GLUT4 in the Endocrine Pancreas Indicating an Impact in Pancreatic Islet Cell Physiology?

Size: px
Start display at page:

Download "GLUT4 in the Endocrine Pancreas Indicating an Impact in Pancreatic Islet Cell Physiology?"

Transcription

1 442 Originl Bsic in the Endocrine Pncres Indicting n Impct in Pncretic Islet Cell Physiology? Authors I. Bähr 1, I. Bzwinsky-Wutschke 1, S. Wolgst 2, K. Hofmnn 1, S. Streck 1, E. Mühluer 2, D. Wedekind 3, E. Peschke 1 Affilitions 1 Institute of Antomy nd Cell Biology, Mrtin Luther University Hlle-Wittenerg, Hlle/Sle, Germny 2 Sxon Acdemy of Sciences Leipzig, Leipzig, Germny 3 Institute of Lortory Animl Science, Hnnover Medicl School, Hnnover, Germny Key words pncretic islet dietes insulin glucose Astrct The glucose trnsporter is well known to fcilitte the trnsport of lood glucose into insulin-sensitive muscle nd dipose tissue. In this study, moleculr, immunohistochemicl, nd Western lot investigtions reveled evidence tht is lso locted in the mouse, rt, nd humn endocrine pncres. In ddition, high glucose decresed nd insulin elevted the expression in pncretic α-cells. In contrst, high glucose incresed expression, wheres insulin led to reduced expression level of the glucose trnsporter in pncretic β-cells. In vivo experiments showed tht in pncretic tissue of type 2 dietic rts s well s type 2 dietic ptients, the expression is significntly incresed compred to the nondietic control group. Furthermore, type 1 dietic rts exhiited reduced trnscript levels in pncretic tissue, wheres insulin tretment of type 1 dietic nimls enhnced the expression ck to control levels. These dt provide evidence for the existence of in the endocrine pncres nd indicte physiologicl relevnce of this glucose trnsporter s well s chrcteristic chnges in dietic disese. received ccepted Biliogrphy DOI /s Pulished online: April 5, 212 Horm Met Res 212; 44: Georg Thieme Verlg KG Stuttgrt New York ISSN Correspondence Dr. I. Bähr Institute of Antomy nd Cell Biology Mrtin Luther University Hlle-Wittenerg Große Steinstrße Hlle/Sle Germny Tel.: +49/345/ /341/ Fx: +49/345/ in.ehr@medizin.uni-hlle.de Introduction Cells in the humn ody use glucose s their mjor source of energy. To ensure constnt supply of glucose, it is essentil tht lood glucose levels re mintined t reltively constnt levels. In the cse of hypoglycemic conditions, the hormone glucgon is secreted y pncretic α-cells, which enhnces lood glucose levels minly y ctivtion of the heptic glycogenolysis nd gluconeogenesis. In contrst, lood glucose levels rise during the fed or sorptive stte, the glucgon secretion is inhiited, nd pncretic β-cells respond y secreting insulin. Insulin cuses decrese in lood glucose ck to norml levels y severl effects: incresing the cellulr glucose influx rte, stimultion of the glycogen synthesis in liver nd skeletl muscle cells, enhncing the ft synthesis in liver, nd dipose tissue s well s fcilittion of the glucose uptke from the lood strem in severl cells. The lst-mentioned effect the insulin-stimulted glucose uptke is primrily medited y the glucose trnsporter isoform (GLUT) 4, which is predominntly expressed in skeletl nd crdic muscle s well s in dipose tissue. In the sl stte, is locted in smll storge vesicles throughout the cytoplsm nd the perinucler region of the cell. Insulin induces the trnsloction of from intrcellulr cytoplsmtic storge sites to the plsm memrne vi inding to the insulin receptor nd, susequently, ctivtion of the insulin receptor sustrte (IRS-1), the phosphtidylinositol 3-kinse (PI3K), nd the Akt kinse signling pthwy [ 1 ]. The plsm memrne-loclized trnsporters permit the fcilitted influx of glucose into the cell, leding to intrcellulr glycolysis or glycogen synthesis. is thus mjor meditor of glucose removl from the circulting lood strem nd therefore key fctor in regultion of the lood glucose homeostsis y insulin. It is well known tht type 1 nd type 2 dietes goes long with reduction of mrna expression nd protein level s well s trnsloction in muscle, crdic nd dipose tissue of different niml models [2 4 ] s well s in humn ptients [5 7 ]. Mice lcking the receptor revel insulin resistnce, impired glucose metolism nd develop type 2 dietes [8, 9 ]. Moreover, overexpression of in knockout mice normlizes insulin sensi- Bähr I et l. in the Endocrine Pncres Horm Met Res 212; 44:

2 Originl Bsic 443 tivity nd glucose tolernce [1, 11 ]. Severl fctors re discussed to e involved in the reltionship etween dysregultion nd the development of insulin resistnce or type 2 dietes. On the one hnd, type 2 dietic s well s oese sujects revel reduced insulin-stimulted IRS-1 tyrosine phosphoryltion nd decresed PI3K ctivity comined with n impired glucose trnsport [12 14 ]. In ddition, overexpression of proteins tht inhiit the insulin signling pthwys, like protein kinse C, my led to n impired insulin-stimulted glucose uptke [15, 16 ]. On the other hnd, chnges in memrne or cytoskeletl physiology of muscle cells or dipocytes s well s circulting fctors, like serum retinol inding protein, free ftty cids or tumor necrosis fctor α, re components influencing the insulin-regulted glucose trnsport [ ]. Besides the high expression of in dipose, skeletl nd crdic muscle tissue, the insulin-dependent glucose trnsporter is locted in the rin nd the kidney in much lower levels [ 22, 23 ], ut could only rrely e detected in other tissues, like the intestine nd the liver [ 24 ]. The im of this study ws to investigte the existence of in mouse, rt, nd humn pncretic tissue including glucose- or insulin-stimulted pncretic islet α- nd β-cell lines for the first time. In ddition, differences of the pncretic mrna expression in type 2 dietic rts nd ptients s well s type 1 dietic nd insulin-treted nimls were evluted. Mterils nd Methods Chemicls nd medi Insulin nd glucose solution, Dulecco s Modified Egle s Medium (DMEM), ovine serum lumin (BSA) s well s protese nd phosphtse inhiitor cocktil were purchsed from Sigm-Aldrich Chemie GmH (Tufkirchen, Germny). Fetl ovine serum (FBS), Roswell Prk Memoril Institute medium (RPMI 164), penicillin, streptomycin, L -glutmine, HEPES, nd nonessentil mino cids were cquired from Biochrom AG (Berlin, Germny). Cell culture The mouse glucgon-producing α-cell line αtc1 clone 9 (αtc1.9) ws purchsed from the Americn Type Culture Collection (Mnsss, VA, USA). Cells were routinely cultured in DMEM with 4 mm L -glutmine, 16.7 mm glucose, 15 mm HEPES, 1.5 g/l sodium icronte,.1 mm nonessentil mino cids,.2 % BSA, 1 % FBS, 1 IU/ml penicillin, nd 1 μm streptomycin. The glucose-responsive, insulin-secreting β-cell line INS1 ws kindly provided y Prof. Wollheim, Deprtment of Medicine, University of Genev, Switzerlnd. INS1 cells were cultured in RPMI 164 medium contining 11.1 mm of glucose supplemented with 1 % FBS, 1 mm HEPES, 2 mm L -glutmine, 1 mm sodium pyruvte, 5 μm 2-mercptoethnol, 1 IU/ml penicillin, nd 137 μm streptomycin. All cells were incuted t 37 C nd 5 % CO 2 nd were split once week in 1:4 fshion. Cell incution experiments To study the mrna expression of glucose trnsporters in pncretic α- nd β-cells fter insulin tretment nd under hyperglycemic conditions, αtc1.9 nd INS1 cells were seeded into 24-well cell culture pltes using DMEM for oth cell lines. After 36 h, cells were wshed with fresh medium. Susequently, cells were incuted with DMEM contining 5.6 mm nd 16.7 mm glucose or in the sence or presence of insulin (1 nm) t 37 C for 16 h. At the end of the incution period, cells were collected nd frozen t 8 C until extrcted. Humn mteril Surgicl smples of pncretic tissue were otined from 28 ptients (15 femles, 13 mles; ged etween 28 nd 78 yers), who underwent prtil or totl pncretectomy ecuse of tumors in pncretic tissue, crcinom of the ppill Vteri, gstric or duodenl crcinom, ile duct crcinom, or severe chronic pncretitis. 7 of these ptients suffered from type 2 dietes. Dt of the ptients records indicte no significnt vritions in mediction or other complictions etween type 2 dietic nd control ptients. Pncretic tissue free of tumor ws otined from dignostic mteril of the Deprtment of Pthology of the Mrtin Luther University Hlle-Wittenerg [ 25 ]. The pncretic mteril ws immeditely immersed in RNAlter (Amion, Austin, TX, USA) until extrction. The permission from the ethics commission of the Fculty of Medicine of the Mrtin Luther University Hlle-Wittenerg ws otined. Additionlly, totl RNA of humn pncres gined from nondietic person ws used s control tissue (Amion, Austin, TX, USA). Totl RNA from islets of Lngerhns, isolted from helthy humn pncres s control tissue, ws ville from SBI (System Biosciences, Mountin, CA, USA). Tissue of humn pectorl muscle ws otined from femle ody donted to the Deprtment of Antomy nd Cell Biology, Mrtin Luther University Hlle-Wittenerg. The muscle tissue ws dissected within time-frme of 4 12 h post mortem. The pproch ws pproved y Institutionl Review Bord regultions, informed consent regultions, nd the provisions of the Declrtion of Helsinki. Animls nd tissue smpling To study the pncretic mrna expression in dietic nimls, type 2 dietic Goto-Kkizki (GK) rts (inred; Tconic M & B, Ry, Denmrk) s well s nondietic Wistr (WR) rts (outred; Schönwlde, Germny) s control group were used. GK rts re nonoese strin tht ws estlished s polygenic model of type 2 dietes. The nimls exhiit mild hyperglycemi, decresed β-cell mss, impired glucose-induced insulin secretion, nd glucose intolernce nd peripherl insulin resistnce [26 ]. In ddition, type 1 dietic LEW.1AR1- iddm rts s well s normoglycemic LEW.1AR1 rts were otined from the Institute of Lortory Animl Science, Hnnover (Medicl School, Hnnover, Germny). The LEW.1AR1- iddm rt is n niml model of utoimmune type 1 dietes with spontneous disese mnifesttion round dy 6 of life. In dietic nimls, pncretic β-cells in the immune-cell infiltrted pncretic islets re quickly destroyed leding to lck of insulin secretion ssocited with hyperglycemi [ 27 ]. Dietic nimls rpidly develop the typicl phenotype of type 1 dietes nd die not lter thn 1 dys fter mnifesttion if left untreted. Furthermore, rts with mnifested type 1 dietes were treted with insulin y sucutneous insulin pellet implnttion (Linplnt, LinShin Inc., Toronto, Cnd) for out 1 dys, s descried erlier [ 28 ]. All nimls were mle nd t the ge of 8 week. They were mintined in groups of 3 nimls per cge nd sujected to light regime of 12-h light nd 12-h drk with light on t 6: h nd controlled temperture of 22 ± 1 C. A stndrd diet ws fed (Altromin 1324; Altromin, Lge, Germny) nd wter ws ccessile d liitum. Rts were scrificed y hert ventricle puncture Bähr I et l. in the Endocrine Pncres Horm Met Res 212; 44:

3 444 Originl Bsic Tle 1 Oligonucleotide primers used in rel-time reverse trnscriptse polymerse chin rection for specific gene mplifiction of glucose trnsporters (GLUT). Genes Product length Primer sequence (forwrd) Primer sequence (reverse) β-actin 389 p 5 -ACTCCTACGTGGGCGACGAGG-3 5 -CAGGTCCAGACGCAGGATGGC-3 (mouse) 269 p 5 -GTCCTGGGGGACCGGATTCCAT-3 5 -CCCTGATGTTAGCCCTGAGTAGGCG-3 (rt) 259 p 5 -GGGCCTGCCCGAAAGAGTCT-3 5 -AGGCTGGCTGTTCCACCCCA-3 (humn) 192 p 5 -CTACCCGGCCCAGAGACCCC-3 5 -TGGCCACGGCCAAACCACAA-3 GLUT1 (mouse) 244 p 5 -AAAGAAGAGGGTCGGCAGAT-3 5 -ACAGCGACACCACAGTGAAG-3 GLUT2 (rt) 183 p 5 -TGGGTTCCTTCCAGTTCG-3 5 -AGGCGTCTGGTGTCGTATG-3 while under deep nesthesi. Muscle nd pncretic tissue were removed; the pncret were immeditely immersed in RNAlter (Amion) nd stored t 2 C. Muscle nd liver tissues were snp-frozen in liquid nitrogen nd stored t 8 C until exmined. Smples of islets of Lngerhns s well s exocrine pncretic tissue from WR rts were collected s descried erlier [ 29 ]. Moleculr nlysis Totl RNA of humn muscle nd pncret, rt muscle, pncres, islets nd INS1 cells s well s of αtc1.9 cells were extrcted using Trizol-sed extrction method ccording to mnufcturer s instructions (PeqGOLD TriFst TM, Peql Biotechnologie GMBH, Erlngen, Germny). The RNA ws quntified spectrophotometriclly nd its integrity ws checked y grose gel electrophoresis. To remove contminting DNA from RNA smples, DNse tretment ws performed (DNA-free TM, Amion Inc., Drmstdt, Germny). Trnscription of the single-strnded RNA into cdna ws chieved y reverse trnscriptse rection ccording to the supplier s instruction (Promeg, Mdison, WI, USA). Smples of mouse muscle, pncres nd islet cdna were ville from former studies [ 3 ]. Rel-time polymerse chin rections (RT-PCR) were performed s formerly descried [ 3 ]. Primers nd sequences re listed in Tle 1. For normliztion of trget gene vlues, β-ctin mrna expression ws used s reference throughout the experiments. After mplifiction, PCR products were seprted y electrophoresis in 3-( N-morpholino) propnesulfonic cid (MOPS)-uffered 1.5 % grose gel, stined with ethidium romide, nd visulized under UV light. Specific mplifiction products were confirmed y restriction digestion ccording to the mnufcturer s instructions (Ferments, St. Leon-Rot, Germny). Immunohistochemicl procedures The pncret of ptients nd wild-type mice s well s skeletl muscle nd liver smples of Wistr rts (n = 4) were immeditely fixed y immersion in 4 % prformldehyde in phosphte-uffered sline (ph 7.4) nd finlly emedded in prffin. Seril sections (5 μm) were cut on microtome nd mounted on glss slides. INS1 cells nd αtc1.9 cells were grown directly onto coverslips nd treted s descried erlier [ 31 ]. Bsiclly ccording to protocol of Bzwinsky-Wutschke et l. [ 31 ] single- or doule-immunofluorescence-leling methods were performed on pncretic tissues nd cultured cells. The following specific primry ntiodies were used: rit nti- (1:1, Millipore GmH, Germny) nd mouse nti-glucgon (1:1, monoclonl, clonek79b1, Sigm-Aldrich). In control sections, the primry ntiody ginst ws replced y norml got serum to test the specificity of the secondry ntiody. Anlyses were performed with confocl lser scnning microscope Leic TCS SP (Leic, Wetzlr, Germny) equipped with the pproprite lsers nd filter sets. Z-series of 12 it were recorded using 4 oil-immersion ojective nd zoom fctor 2.5. The imges of Z-series were comined into single imge using n pproprite softwre progrmme. Western lot experiments For Western lot nlyses, skeletl muscle, liver, nd pncretic tissue otined from LEW.1AR1 rts, skeletl muscle otined from wild-type mice s well s INS1 cells, nd αtc1.9 cells were used. Totl protein ws otined s previously descried [ 31 ] y incution of the tissue of rt or mouse or cells with RIPA lysis uffer (contining 5 mm Tris, 1 % IGEPAL C-63,.5 % sodium deoxycholte,.1 % sodium dodecyl sulfte (SDS), protese nd phosphtse inhiitor cocktil) on ice for 3 min followed y soniction nd further incution on ice. After centrifugtion of the homogente t 11 g for 2 min, the superntnt ws mixed with n equl mount of 2 smple uffer (contining 1 mm Tris, 2 % glycerol, 4 % SDS, nd 4 % mercptoethnol). Totl protein (25 μg) ws seprted y SDS-polycrylmide gel electrophoresis nd trnsferred onto PVDF memrne (Crl Roth GmH & Co. KG, Krlsruhe, Germny) using stndrd protocols. Memrnes were locked with 5 % BSA in Tris-uffered sline nd incuted with primry ntiody detecting (Millipore) t 1:2 dilution. Horserdish peroxydse-conjugted secondry ntiody ws employed t 1:1 dilution. Enhnced chemiluminescence detection (SuperSignl, Pierce, Rockford, Il, USA) ws used to visulize immunorective nds. Sttisticl nlysis Sttisticl nlyses were performed using GrphPd softwre (GrphPd Softwre Inc., L Joll, CA, USA). For sttisticl evlution nd significnce testing of differences, the Mnn-Whitney U-test ws performed. p-vlues of <.5 were considered significnt. Dt were presented s men ± SEM of given smple numer. Results Detection of mrna nd protein in rt, mouse, nd humn pncretic tissue Moleculr investigtions reveled evidence tht the glucose trnsporter is expressed in pncretic tissue of rt nd mouse. RT-PCR rections were performed using specific primers ( Tle 1 ). Gel electrophoresis visulized the specific mplifiction product of rt t 259 p ( Fig. 1 ) nd mouse t 269 p ( Fig. 1e ) in whole pncretic tissue nd islets of Lngerhns s well s in muscle s positive control tissue. In prticulr, mrna expression could e estlished in the rt pncretic β-cell line INS1 ( Fig. 1 ) nd the Bähr I et l. in the Endocrine Pncres Horm Met Res 212; 44:

4 Originl Bsic p 3 p 2 p c e 4 p 3 p 2 p 5 µm Rt pncretic tissue GLUC 5 µm Mouse pncretic tissue f 3 p 2 p 15 p 1 p 75 p GLUC/ 5 µm 3 p 2 p 15 p LR P R d LR P R 2 µm Fig. 1 Evidence for the glucose trnsporter in the pncretic tissue of rts d nd mice e h. Moleculr nlysis reveled the expression of mrna in muscle, pncres, exocrine pncretic tissue, nd islets of Lngerhns of rts (, 259 p) nd mice ( e, 269 p) s well s expression of the mrna in rt INS1 cells nd mouse αtc1.9 cells e. Restriction rection using the restriction enzymes BsrI (, rt) nd LweI ( f, mouse) resulted in defined frgments of the mplifiction products with 197 p nd 62 p moleculr size (rt) or 115 p nd 154 p moleculr size (mouse) c, g. Immunohistochemicl stining of (red) is shown to e concentrted predominntly in rt c or mouse g pncretic islets nd distriuted within the islet. Leling of glucgon (GLUC, green) nd merged signls indicte co-locliztion of glucgon nd s well s protein expression in oth α- nd β-cells. d, h Immunopositive leling of INS1 cells d nd αtc1.9 cells h with. L: 1-p ldder; LR: low-rnge ldder; NTC: nontemplte control; P: mplifiction product; R: restriction frgments. 1 p g h GLUC GLUC/ 5 µm 5 µm 5 µm 2 µm mouse pncretic α-cell line αtc1.9 ( Fig. 1e ). In exocrine pncretic tissue, no mrna expression ws detected neither in rt nor in mouse pncres. The identity of the mplifiction products in INS1 cells nd αtc1.9 cells ws confirmed y restriction nlysis. Incution with the restriction enzyme BsrI led to frgments of the rt trnscript (259 p) with 197 p nd 62 p ( Fig. 1 ). The mouse trnscript (269 p) ws cut into 2 specific frgments with 154 p nd 115 p using the restriction enzyme LweI ( Fig. 1f ). Restriction rections were lso successfully performed in muscle tissue of mice nd rts (dt not shown). Immunohistochemicl leling of rt or mouse pncretic tissue showed specific leling of predominntly in pncretic islets. Only wek immunofluorescence ws detected in the exocrine tissue surrounding the islets ( Fig. 1c, g ). immunoleling exhiited dense punct-like distriution pttern in pncretic islets, which excluded the cell nuclei. Furthermore, immunostining indictes tht is locted in the rt β-cell line INS1 nd α-cell line αtc1.9 ( Fig. 1d, h ), which corresponds to the detection of mrna in oth cell lines y RT- PCR. In ddition, co-leling with glucgon specific ntiody s well s the merging of nd glucgon signls were performed to revel co-locliztion nd to differentite etween α- nd β-cells s the mjor islet cell types in rt nd mouse pncretic islets ( Fig. 1c, g ). The findings demonstrte tht pncretic α-cells s well s β-cells exhiit the glucose trnsporter. Beside the detection of in niml pncretic tissue, the glucose trnsporter ws lso reveled in smples of humn pncretic tissue. As demonstrted in Fig. 2, -specific mplifiction product (192 p) is expressed in whole pncres, islets of Lngerhns s well s in humn muscle s positive control tissue. Restriction rection of the humn product in islets of Lngerhns resulted in specific frgments with 15 p, 52 p, nd 35 p ( Fig. 2 ) using the restriction enzyme PstI. Restriction rections were lso successfully performed in humn muscle tissue (dt not shown). Immunohistochemicl nlysis indicted the predominnt locliztion of in the humn pncretic islet ( Fig. 2c ), wheres exocrine tissue showed wek leling. As reveled y glucgon co-leling, is expressed in oth α- nd β-cell types in humn islets ( Fig. 2c ). To demonstrte the specificity of the ntiody, severl control nlyses were performed ( Fig. 3 ). Besides the detection of in rt pncretic islets y immunohistochemistry, immunostining ws confirmed in rt skeletl muscle tissue s positive control smple. Moreover, no immunorection ws otined in smples of rt liver, known negtive control In ddition, negtive control slides processed without primry ntiody were included for ech stining to rule out nonspecific inding ( Fig. 3 ). To prove ntiody specificity y n lterntive technique, Western lot nlyses using the sme ntiody s in immunohistochemistry lso confirmed protein expression in rt nd mice pn- Bähr I et l. in the Endocrine Pncres Horm Met Res 212; 44:

5 446 Originl Bsic 4 p 3 p 2 p c Humn pncretic tissue GLUC LR P R 3 p 2 p 15 p 1 p 75 p 5 p GLUC/ Fig. 2 Evidence for the glucose trnsporter in humn pncretic tissue. Moleculr nlysis reveled the expression of mrna (192 p) in humn muscle, pncres of control, nd ptient tissue s well s islets of Lngerhns. Restriction rection using the restriction enzyme PstI resulted in frgments of the mplifiction product with 15 p, 52 p nd 35 p (smll product not shown). c Humn pncretic islets show strong immunoleling (red) exhiiting ring-like structures. Leling with glucgon (GLUC, green) nd merged signls indicte co-locliztion of glucgon nd s well s protein expression in oth α- nd β-cells. L: 1-p ldder; LR: low-rnge ldder; NTC: nontemplte control; P: mplifiction product; R: restriction frgments. 5 µm 5 µm 5 µm cretic tissue s well s in INS1 nd αtc1.9 cells ( Fig. 3 ). In ccord with the immunohistochemicl investigtions, signls were otined lso in Western lots using skeletl muscle tissue (positive control) ut not with liver tissue (negtive control). Influence of glucose nd insulin on mrna expression of glucose trnsporters in INS1 cells nd αtc1.9 cells To investigte whether the level of mrna expression in pncretic β-cells cn e influenced y high glucose concentrtions or insulin, INS1 cells were incuted under hyperglycemic conditions or with insulin for 16 h. High glucose levels of 16.7 mm resulted in significnt elevtion of the mrna expression for nerly 1 % compred to the control cells incuted under normoglycemic conditions ( Fig. 4 ), wheres no chnges in GLUT2 expression were detected under different glucose concentrtions ( Fig. 4c ). Incution with insulin (1 nm) nd 5.6 mm glucose concentrtion led to significntly reduced nd GLUT2 mrna expression levels compred to untreted control INS1 cells ( Fig. 4, d ). In ddition to effects in INS1 cells, the influence of high glucose or insulin incution on mrna expression in pncretic α-cells ws investigted. Incution of αtc1.9 cells under hyperglycemic conditions with 16.7 mm glucose in the medium resulted in significntly reduced mrna expression levels compred to control cells incuted t 5.6 mm glucose ( Fig. 4f ). Expression levels of GLUT1 mrna in αtc1.9 cells remined unffected under different glucose concentrtions ( Fig. 4h ). Moreover, mrna expression ws significntly enhnced fter insulin tretment (1 nm) for out 6 % of the control cells ( Fig. 4g ), wheres GLUT1 mrna expression ws only slightly, ut not significntly incresed fter insulin incution ( Fig. 4i ). Furthermore, the rtios of glucose trnsporter expression were nlyzed in INS1 s well s in αtc1.9 cells ( Fig. 4e, j ). The expression levels of oth the GLUT2 mrna in INS1 cells ( Fig. 4e ) nd the GLUT1 mrna in αtc1.9 cells ( Fig. 4j ) were out 3 times higher compred to mrna expression levels in the respective cell line. Pncretic mrna expression in type 2 dietic rts nd ptients It is well known tht type 2 dietes is ssocited with disturnces of pncretic islet cell physiology s well s with reduced mrna expression levels nd dysfunction of the glucose trnsporter in dipose nd muscle tissue. To investigte whether the pncretic mrna expression is lso ffected in type 2 dietes, rel-time RT-PCR nlysis ws performed using pncretic mrna of control Wistr (WR) rts nd type 2 dietic Goto-Kkizki (GK) rts. Interestingly, the mrna level ws significntly incresed in pncret of GK rts compred to the nondietic WR control group ( Fig. 5 ). In ddition to the niml studies, investigtions on pncretic tissue of nondietic nd type 2 dietic ptients were executed. The results demonstrte tht the pncretic mrna expression ws significntly elevted in type 2 dietic ptients ( Fig. 5 ). Pncretic mrna expression in type 1 dietic nd insulin-treted rts Besides exmining type 2 dietic rts nd ptients, pncretic tissue of type 1 dietic nimls ws chrcterized ccording to the mrna expression sttus. Therefore, LEW.1AR1- iddm rts s n niml model of spontneous insulin-dependent dietes mellitus s well s the nondietic LEW.1AR1 rts were used. As the dt show, pncretic mrna expression is significntly reduced in type 1 dietic LEW.1AR1- iddm rts compred to the LEW.1AR1 control group ( Fig. 5c ). Astonishingly, insulin-sustitution of LEW.1AR1- iddm rts suffering from type 1 dietes normlized the pncretic mrna expression levels nerly to those of nondietic control rts. Discussion The glucose trnsporter ws descried to e exclusively present in dipose, crdic nd muscle tissue mediting n insulin-induced stimultion of glucose trnsport into cells. Moleculr, immunohistochemicl nd Western lot investigtions of this study reveled evidence tht is lso locted in mouse, Bähr I et l. in the Endocrine Pncres Horm Met Res 212; 44:

6 Originl Bsic 447 GLUT 4 Pncres Liver Muscle Negtive control 5 µm 5 µm 5 µm 5 µm 5 µm 5 µm MW (kd) Fig. 3 Anlyses of the ntiody specificity. Immunohistochemicl stining of ws performed in rt pncres s well s in rt liver s negtive control tissue nd rt skeletl muscle s positive control tissue. sections were incuted under the sme conditions ut using norml got serum insted of the primry ntiody. Western lot experiments using the ntiody demonstrte protein expression in rt nd mice pncretic tissue s well s INS1 nd αtc1.9 cells. Skeletl muscles were used s positive control smples nd rt liver tissue s negtive control smples lcking the signl. rt, nd humn pncretic tissue with predominnt expression in islets of Lngerhns. In the pst, only one study could demonstrte the existence of in humn pncretic tissue [ 32 ]. In concordnce with our results, the mrna expression s well s protein level of ws detected nd immunostining of protein in humn pncres reveled prevlent loction in the endocrine pncres. However, there re contrry results of others where the detection of in rt islets hd filed [ 33 ]. In ddition to exmintions on pncretic islets, we lso detected the directly in the β-cell line INS1, which confirms results of n erlier study on the β-cell line β-tc [33 ]. Furthermore, we re the first to revel expression in pncretic α-cell line. Chnges in mrna expression fter glucose or insulin incution indicte functionl relevnce of this glucose trnsporter for the α- nd β-cell physiology. Insulin reduced the mrna expression in INS1 cells nd therefore supposly the glucose influx in β-cells, possily leding to negtive feedck signl on insulin secretion. High expression under hyperglycemic conditions is indictive for n insulin-incresing signl due to elevted glucose uptke rtes in β-cells. Surprisingly, glucose nd insulin incution led to opposite effects on mrna expression in αtc1.9 cells. These findings re interesting s glucgon secretion from pncretic α-cells cts s n opponent of insulin leding to rise of lood glucose levels in terms of hypoglycemi. Glucgon relese is inhiited y high glucose concentrtions s well s y insulin [ 34 ]. Insulin medites its glucgon-inhiiting effects y reducing the sensitivity of ATP-dependent potssium chnnels vi the PI3K signling cscde [35, 36 ] or y trnsloction of GABA-A receptors vi n Akt kinse-dependent pthwy [37 ]. The results of this study showed tht insulin incution elevted mrna expression levels in αtc1.9 cells which my e ssocited with n increse of glucose influx in these cells nd consequently reduction of glucgon secretion. Thus, the dt indicte tht insulin medites its inhiiting effect on α-cells y modulting glucose uptke vi. In contrst, it remins uncler why high glucose resulted in reduction of mrna expression in αtc1.9 cells, s reduced glucose influx would rther e ccompnied y stimultion of glucgon relese. Possily, this pthwy is feedck mechnism preventing n excessive reduction of glucgon secretion under hyperglycemic conditions. Our findings of chnges in expression in INS1 nd αtc1.9 cells under hyperglycemic conditions or fter insulin tretment re sed on investigtions of mrna expression levels. Future investigtions nlyzing protein mount, cellulr loction of the glucose trnsporter s well s glucose uptke experiments re necessry to confirm the results. However, recent studies demonstrted tht incresed mrna is ssocited with elevted protein levels nd enhnced glucose uptke rtes in skeletl muscle nd dipocytes [38 4 ] leding to the ssumption tht this correltion lso pertins for pncretic α- nd β-cells. The occurrence of other glucose trnsporter isoforms is well chrcterized in pncretic islet cells. The most importnt glucose trnsporter in β-cells is the insulin-independent GLUT2, which trnsports extrcellulr glucose vi fcilitted diffusion with high trnsport cpcity, leding to exocytosis of insulin [41 ]. In pncretic α-cells, glucose influx is medited vi GLUT1, which is known to e responsile for the low-level of sl glucose uptke [ 42 ]. Our incution studies showed, tht in contrst to the trnscript the GLUT2 mrna expression in INS1 cells s well s the GLUT1 mrna expression in αtc1.9 cells were unffected under hyperglycemic conditions. However, insulin tretment influenced oth the trnscript level s well s the GLUT1 mrna expression in αtc1.9 cells nd GLUT2 mrna expression in INS1 cells. These findings indicte tht insulin hs regultory effects on different glucose trnsporters in pncretic islet cells. But it hs to e considered tht in oth cell types used in these investigtions, the mrna expression of the descried prticulr glucose trnsporter ws out 3 times higher thn the detected mrna level. Therefore, the functionl relevnce of GLUT1 or GLUT2 in glucose trnsport in pncretic α- nd β-cells seems to e the most importnt, ut oviously, our results revel regultory mening of in these islet cells. However, further studies, like glucose uptke mesurements or up- nd downregultion of mrna in α- nd β-cell lines, re prospectively necessry to investigte how much glucose ctully enters α- or β-cells vi the different glucose trnsporter isoforms nd whether the glucose influx Bähr I et l. in the Endocrine Pncres Horm Met Res 212; 44:

7 448 Originl Bsic Reltive expression c Reltive GLUT2 expression 25 *** mm 16.7 mm Glucose 5.6 mm 16.7 mm Glucose Reltive expression d Reltive GLUT2 expression 15 *** INS1 cells 15 * Insulin (1 nm) Insulin (1 nm) e Reltive expression of glucose trnsporter GLUT2 *** Fig. 4 I nfluence of glucose or insulin on the mrna expression of the glucose trnsporter GLUT2 nd in INS1 cells d nd of the glucose trnsporter GLUT1 nd in αtc1.9 cells f i. INS1 cells or αtc1.9 cells were incuted with 5.6 mm or 16.7 mm glucose in medium, c, f, h or with or without insulin (1 nm) t 5.6 mm glucose concentrtion, d, g, i for 16 h. e, j Expression rtios of glucose trnsporters in INS1 nd αtc1.9 cells: Rtio of GLUT2 vs. mrna expression in INS1 cells e nd rtio of GLUT1 vs. mrna expression in αtc1.9 cells j. Rel-time RT-PCR ws performed to quntify the GLUT1, GLUT2, nd mrna expression. The expression of trget gene trnscripts ws clculted reltive to β-ctin mrna levels s reference. Results were expressed s percentge of the untreted controls nd s mens ± SEM of 3 individul experiments, ech with n = 4 tches. Asterisks indicte significnt differences t *p <.5 nd ***p <.1. f Reltive expression h Reltive GLUT1 expression * 5.6 mm 16.7 mm Glucose 5.6 mm 16.7 mm Glucose g Reltive expression i Reltive GLUT1 expression αtc1.9 cells *** Insulin (1 nm) Insulin (1 nm) j Reltive expression of glucose trnsporter GLUT1 *** into pncretic islet cells cn e sustntilly modulted y the. Moreover, eside the existence of in endocrine pncretic cells, wek immunoflourenscence signls lso indicte expression in the exocrine pncretic tissue. The possile mening nd function of this glucose trnsporter in exocrine pncretic cells remin to e elusive. In ddition to investigtions on different cell lines, mrna expression in pncretic tissue of type 1 nd type 2 dietic nimls s well s on type 2 dietic ptients were performed. In pncretic tissue of type 2 dietic rts nd ptients, mrna expression ws significntly elevted. These results were surprising s in muscle, crdic nd dipose tissue the mrna expression ws shown to e reduced [2, 3, 6, 7 ] or not ffected [43, 44 ] in type 2 dietic stte. Nevertheless, our findings of enhnced mrna expression in pncres were confirmed oth in rt nd even in humn tissue. As demonstrted in previous studies, the pncretic GLUT2 trnscript level is significntly reduced in type 2 dietic rts nd mice [45, 46 ]. Possily, the up regultion of pncretic trnscript levels is physiologicl effect to compenste low GLUT2 concentrtions in the type 2 dietic stte. In contrst, pncretic mrna expression is downregulted in insulin-deficient type 1 dietic LEW.1AR1- iddm rts, s it ws lso shown in severl studies regrding expression in skeletl muscle of type 1 dietic nimls treted with lloxn or streptozotocin [47 5 ]. Interestingly, insulin tretment of LEW.1AR1- iddm rts cused recovery of pncretic mrna levels nerly to those oserved in untreted control nimls. Therefore, the direct ssocition etween insulin nd expression in pncretic tissue could e reveled in vivo for the first time. The type 1 dietic LEW.1AR1- iddm rts develop spontneous insulin-dependent utoimmune dietes through β-cell poptosis, wheres other islet cell types re not ltered [27 ]. Therefore, the enhnced mrna expression in pncret of type 1 dietic nimls fter insulin tretment cnnot e referred to n effect in β-cells, ut supposly in α-cells, s incution of αtc1.9 cells with insulin lso incresed the mrna expression levels. Unfortuntely, due to the effects of type 1 nd 2 dietes on pncretic mrna expression, we were not exctly le to distinguish etween exocrine nd endocrine pncres frction. Nevertheless, moleculr nd immunohistochemicl nlyses Bähr I et l. in the Endocrine Pncres Horm Met Res 212; 44:

8 Originl Bsic 449 Reltive expression WR rts ** c 1 Reltive expression Type 2 dietic GK rts Reltive expression *** *** ptients * Type 2 dietic ptients Fig. 5 Expression of the glucose trnsporter mrna in pncret of nondietic nd type 2 dietic rts nd ptients s well s of type 1 dietic nd insulin-treted rts. Reltive mrna expression levels in pncretic tissue of control Wistr (WR) rts nd type 2 dietic Goto- Kkizki (GK) rts (n = nimls). Reltive mrna trnscript levels in pncret of nondietic ptients s well s of type 2 dietic ptients [n = 7 (type 2 dietic) or 22 (nondietic control group)]. c Reltive expression of the glucose trnsporter mrna in pncret of nondietic LEW.1AR1 (LEW) rts, type 1 dietic LEW.1AR1- iddm (IDDM) rts nd insulin-treted, type 1 dietic LEW.1AR1- iddm rts (n = nimls). The expression of trnscripts ws clculted reltive to β-ctin mrna levels. Vlues re expressed s mens ± SEM. Asterisks indicte significnt differences t *p <.5; **p <.1 nd ***p <.1. LEW rts Type 1 dietic IDDM rts Type 1 dietic IDDM rts (Insulintreted) reveled evidence for predominnt locliztion in islets of Lngerhns ut not in the exocrine pncretic tissue. Therefore, the findings cn e referred to chnges in mrna expression in islet cells. Whether the insulin nd glucose concentrtions in nimls influence expression primrily in pncretic α- or β-cells or even oth cell types remins elusive. Summrizing the results of our in vitro nd in vivo investigtions, we provided evidence for the existence nd indictions of physiologicl relevnce of in the endocrine pncres s well s chrcteristic chnges in dietic disese. The dt suggest tht dietes-ssocited disturnces in function nd glucose uptke re not only reducile to chnges of function in muscle or dipose cells, ut lso of pncretic islet cells. Therefore, pncretic dysfunction my ply role in the development of α- or β-cell dysfunction nd cuse disturnces of the glucose metolism in type 1 nd 2 dietes. Acknowledgements The uthors thnk Ms. Heydel, Ms. Rothgänger, Ms. Jordn, nd Mr. Biemnn for skillful technicl ssistnce nd Ms. Axmnn for linguistic ssistnce. This study ws prtly supported y grnts of the Wilhelm-Roux progrm of the Mrtin Luther University s well s of the Deutsche Dietes-Stiftung. References 1 McCrthy A M, Elmendorf JS. s itinerry in helth & disese. Indin J Med Res 27 ; 125 : Ndisng J F, Lne N, Jdhv A. The heme oxygense system tes hyperglycemi in Zucker dietic ftty rts y potentiting insulinsensitizing pthwys. Endocrinology 29 ; 15 : Desrois M, Sidell RJ, Guguier D, King LM, Rdd GK, Clrke K. Initil steps of insulin signling nd glucose trnsport re defective in the type 2 dietic rt hert. Crdiovsc Res 24 ; 61 : Sivitz W I, DeSutel SL, Kyno T, Bell GI, Pessin JE. Regultion of glucose trnsporter messenger RNA in insulin-deficient sttes. Nture 1989 ; 34 : Wllerg-Henriksson H, Zierth JR. : key plyer regulting glucose homeostsis? Insights from trnsgenic nd knockout mice (review). Mol Memr Biol 21 ; 18 : Gster M, Stehr P, Beck-Nielsen H, Schroder HD, Hnderg A. is reduced in slow muscle fiers of type 2 dietic ptients: is insulin resistnce in type 2 dietes slow, type 1 fier disese? Dietes 21 ; 5 : Armoni M, Hrel C, Br-Yoseph F, Milo S, Krnieli E. Free ftty cids repress the gene expression in crdic muscle vi novel response elements. J Biol Chem 25 ; 28 : Zismn A, Peroni OD, Ael ED, Michel MD, Muvis-Jrvis F, Lowell B B, Wojtszewski JF, Hirshmn MF, Virkmki A, Goodyer L J, Khn C R, Khn B B. Trgeted disruption of the glucose trnsporter 4 selectively in muscle cuses insulin resistnce nd glucose intolernce. Nt Med 2 ; 6 : Pttrnit R, vn den Berg HA, Spnswick D. The development of insulin resistnce in Type 2 dietes: insights from knockout studies. Sci Prog 28 ; 91 : Crvlho E, Kotni K, Peroni OD, Khn BB. Adipose-specific overexpression of reverses insulin resistnce nd dietes in mice lcking selectively in muscle. Am J Physiol Endocrinol Met 25 ; 289 : E551 E Tso T S, Stenit AE, Fctor SM, Chen W, Rossetti L, Chrron M J. Prevention of insulin resistnce nd dietes in mice heterozygous for ltion y trnsgenic complementtion of in skeletl muscle. Dietes 1999 ; 48 : Bjornholm M, Al-Khlili L, Dicker A, Nslund E, Rossner S, Zierth J R, Arner P. Insulin signl trnsduction nd glucose trnsport in humn dipocytes: effects of oesity nd low clorie diet. Dietologi 22 ; 45 : Krook A, Bjornholm M, Glusk D, Jing XJ, Fhlmn R, Myers M G Jr, Wllerg-Henriksson H, Zierth JR. Chrcteriztion of signl trnsduction nd glucose trnsport in skeletl muscle from type 2 dietic ptients. Dietes 2 ; 49 : Bähr I et l. in the Endocrine Pncres Horm Met Res 212; 44:

9 45 Originl Bsic 14 Goodyer L J, Giorgino F, Shermn LA, Crey J, Smith RJ, Dohm GL. Insulin receptor phosphoryltion, insulin receptor sustrte-1 phosphoryltion, nd phosphtidylinositol 3-kinse ctivity re decresed in intct skeletl muscle strips from oese sujects. J Clin Invest 1995 ; 95 : Condorelli G, Vigliott G, Ivrone C, Cruso M, Tocchetti CG, Andreozzi F, Cfieri A, Tecce MF, Formisno P, Beguinot L, Beguinot F. PED/PEA- 15 gene controls glucose trnsport nd is overexpressed in type 2 dietes mellitus. Emo J 1998 ; 17 : Kim J K, Fillmore JJ, Sunshine MJ, Alrecht B, Higshimori T, Kim DW, Liu Z X, Soos T J, Cline G W, O'Brien W R, Littmn DR, Shulmn GI. PKC - thet knockout mice re protected from ft-induced insulin resistnce. J Clin Invest 24 ; 114 : Pilch P F, Thompson PA, Czech MP. Coordinte modultion of D-glucose trnsport ctivity nd ilyer fluidity in plsm memrnes derived from control nd insulin-treted dipocytes. Proc Ntl Acd Sci USA 198 ; 77 : McCrthy A M, Spisk K O, Brozinick J T, Elmendorf J S. Loss of corticl ctin filments in insulin-resistnt skeletl muscle cells impirs vesicle trfficking nd glucose trnsport. Am J Physiol Cell Physiol 26 ; 291 : C86 C Wolf G. Serum retinol-inding protein: link etween oesity, insulin resistnce, nd type 2 dietes. Nutr Rev 27 ; 65 : Boden G. Role of ftty cids in the pthogenesis of insulin resistnce nd NIDDM. Dietes 1997 ; 46 : Hotmisligil G S, Murry DL, Choy L N, Spiegelmn BM. Tumor necrosis fctor lph inhiits signling from the insulin receptor. Proc Ntl Acd Sci USA 1994 ; 91 : El Messri S, Ait-Ikhlef A, Amroise D H, Penicud L, Arluison M. Expression of insulin-responsive glucose trnsporter mrna in the rt rin nd spinl cord: n in situ hyridiztion study. J Chem Neuront 22 ; 24 : Chin E, Zhou J, Bondy C. Antomicl nd developmentl ptterns of fcilittive glucose trnsporter gene expression in the rt kidney. J Clin Invest 1993 ; 91 : Aschench J R, Steglich K, Gel G, Honsch K U. Expression of mrna for glucose trnsport proteins in jejunum, liver, kidney nd skeletl muscle of pigs. J Physiol Biochem 29 ; 65 : Peschke E, Stumpf I, Bzwinsky I, Litvk L, Drlle H, Muhluer E. Meltonin nd type 2 dietes possile link? J Pinel Res 27 ; 42 : Goto Y, Suzuki K, Ono T, Sski M, Toyot T. Development of dietes in the non-oese NIDDM rt (GK rt). Adv Exp Med Biol 1988 ; 246 : Lenzen S, Tiedge M, Elsner M, Lortz S, Weiss H, Jorns A, Kloppel G, Wedekind D, Prokop C M, Hedrich HJ. The LEW.1AR1/Ztm-iddm rt: new model of spontneous insulin-dependent dietes mellitus. Dietologi 21 ; 44 : Peschke E, Hofmnn K, Bhr I, Streck S, Alrecht E, Wedekind D, Muhluer E. The insulin-meltonin ntgonism: studies in the LEW.1AR1- iddm rt (n niml model of humn type 1 dietes mellitus). Dietologi 211 ; 54 : Peschke E, Futeck JD, Musshoff U, Schmidt F, Beckmnn A, Peschke D. Evidence for meltonin receptor within pncretic islets of neonte rts: functionl, utordiogrphic, nd moleculr investigtions. J Pinel Res 2 ; 28 : Muhluer E, Gross E, Lucy K, Wolgst S, Peschke E. Loss of meltonin signlling nd its impct on circdin rhythms in mouse orgns regulting lood glucose. Eur J Phrmcol 29 ; 66 : Bzwinsky-Wutschke I, Wolgst S, Muhluer E, Peschke E. Distriution ptterns of clcium-inding proteins in pncretic tissue of nondietic s well s type 2 dietic rts nd in rt insulinom et-cells (INS-1). Histochem Cell Biol 21 ; 134 : Koyshi H, Mitsui T, Nomur S, Ohno Y, Kdomtsu K, Murmtsu T, Ngsk T, Mizutni S. Expression of glucose trnsporter 4 in the humn pncretic islet of Lngerhns. Biochem Biophys Res Commun 24 ; 314 : Brnt A M, McCoid S, Thoms HM, Bldwin SA, Dvies A, Prker J C, Gis E M, Gould G W. Anlysis of the glucose trnsporter content of islet cell lines: implictions for glucose-stimulted insulin relese. Cell Signl 1992 ; 4 : Kwmori D, Welters HJ, Kulkrni RN. Moleculr pthwys underlying the pthogenesis of pncretic lph-cell dysfunction. Adv Exp Med Biol 21 ; 654 : Leung Y M, Ahmed I, Sheu L, Go X, Hr M, Tsushim RG, Dimnt N E, Gisno H Y. Insulin regultes islet lph-cell function y reducing KATP chnnel sensitivity to denosine 5 -triphosphte inhiition. Endocrinology 26 ; 147 : McDonld P E, De Mrinis YZ, Rmrchey R, Slehi A, M X, Johnson P R, Cox R, Elisson L, Rorsmn P. A K ATP chnnel-dependent pthwy within lph cells regultes glucgon relese from oth rodent nd humn islets of Lngerhns. PLoS Biol 27 ; 5 : e Xu E, Kumr M, Zhng Y, Ju W, Ot T, Zhng N, Liu S, Wendt A, Deng S, Ein Y, Wheeler MB, Brun M, Wng Q. Intr-islet insulin suppresses glucgon relese vi GABA-GABAA receptor system. Cell Met 26 ; 3 : Kern M, Wells JA, Stephens JM, Elton CW, Friedmn JE, Tpscott E B, Pekl P H, Dohm GL. Insulin responsiveness in skeletl muscle is determined y glucose trnsporter (Glut4) protein level. Biochem J 199 ; 27 : Flores-Riveros JR, McLenithn JC, Ezki O, Lne MD. Insulin down-regultes expression of the insulin-responsive glucose trnsporter () gene: effects on trnscription nd mrna turnover. Proc Ntl Acd Sci USA 1993 ; 9 : Kohn A D, Summers SA, Birnum MJ, Roth RA. Expression of constitutively ctive Akt Ser/Thr kinse in 3T3-L1 dipocytes stimultes glucose uptke nd glucose trnsporter 4 trnsloction. J Biol Chem 1996 ; 271 : Efrt S, Tl M, Lodish HF. The pncretic et-cell glucose sensor. Trends Biochem Sci 1994 ; 19 : Heimerg H, De Vos A, Pipeleers D, Thorens B, Schuit F. D ifferences in glucose trnsporter gene expression etween rt pncretic lphnd et-cells re correlted to differences in glucose trnsport ut not in glucose utiliztion. J Biol Chem 1995 ; 27 : Pedersen O, Bk JF, Andersen PH, Lund S, Moller DE, Flier JS, Khn B B. Ev i- dence ginst ltered expression of GLUT1 or in skeletl muscle of ptients with oesity or NIDDM. Dietes 199 ; 39 : Holten M K, Zcho M, Gster M, Juel C, Wojtszewski JF, Del F. St re ng th trining increses insulin-medited glucose uptke, content, nd insulin signling in skeletl muscle in ptients with type 2 dietes. Dietes 24 ; 53 : Frese T, Bzwinsky I, Muhluer E, Peschke E. Circdin nd gedependent expression ptterns of GLUT2 nd glucokinse in the pncretic et-cell of dietic nd nondietic rts. Horm Met Res 27 ; 39 : Chnkiewitz E, Peschke D, Hererg L, Bzwinsky I, Muhluer E, Bromme H J, Peschke E. Did the grdul loss of GLUT2 cuse shift to dietic disorders in the New Zelnd oese mouse (NZO/Hl)? Exp Clin Endocrinol Dietes 26 ; 114 : Zhng Z, Jing J, Yu P, Zeng X, Lrrick JW, Wng Y. Hypoglycemic nd et cell protective effects of ndrogrpholide nlogue for dietes tretment. J Trnsl Med 29 ; 7 : Sto K, Iemitsu M, Aizw K, Ajisk R. DHEA improves impired ctivtion of Akt nd PKC zet/lmd- pthwy in skeletl muscle nd improves hyperglycemi in streptozotocin-induced dietes rts. Act Physiol (Oxf) 29 ; 197 : Hwng S L, Liu IM, Tzeng TF, Cheng JT. Activtion of imidzoline receptors in drenl glnd to lower plsm glucose in streptozotocininduced dietic rts. Dietologi 25 ; 48 : Chn P, Wong KL, Liu IM, Tzeng TF, Yng TL, Cheng JT. Antihyperglycemic ction of ngiotensin II receptor ntgonist, vlsrtn, in streptozotocin-induced dietic rts. J Hypertens 23 ; 21 : Bähr I et l. in the Endocrine Pncres Horm Met Res 212; 44:

EFFECTS OF AN ACUTE ENTERIC DISEASE CHALLENGE ON IGF-1 AND IGFBP-3 GENE EXPRESSION IN PORCINE SKELETAL MUSCLE

EFFECTS OF AN ACUTE ENTERIC DISEASE CHALLENGE ON IGF-1 AND IGFBP-3 GENE EXPRESSION IN PORCINE SKELETAL MUSCLE Swine Dy 22 Contents EFFECTS OF AN ACUTE ENTERIC DISEASE CHALLENGE ON IGF-1 AND IGFBP-3 GENE EXPRESSION IN PORCINE SKELETAL MUSCLE B. J. Johnson, J. P. Kyser, J. D. Dunn, A. T. Wyln, S. S. Dritz 1, J.

More information

Supplementary figure 1

Supplementary figure 1 Supplementry figure 1 Dy 8 post LCMV infection Vsculr Assoc. Prenchym Dy 3 post LCMV infection 1 5 6.7.29 1 4 1 3 1 2 88.9 4.16 1 2 1 3 1 4 1 5 1 5 1.59 5.97 1 4 1 3 1 2 21.4 71 1 2 1 3 1 4 1 5 1 5.59.22

More information

Supplementary Figure 1

Supplementary Figure 1 Supplementry Figure 1 c d Wistr SHR Wistr AF-353 SHR AF-353 n = 6 n = 6 n = 28 n = 3 n = 12 n = 12 Supplementry Figure 1 Neurophysiologicl properties of petrosl chemoreceptive neurones in Wistr nd SH rts.

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION Prentl doi:.8/nture57 Figure S HPMECs LM Cells Cell lines VEGF (ng/ml) Prentl 7. +/-. LM 7. +/-.99 LM 7. +/-.99 Fold COX induction 5 VEGF: - + + + Bevcizum: - - 5 (µg/ml) Reltive MMP LM mock COX MMP LM+

More information

2018 American Diabetes Association. Published online at

2018 American Diabetes Association. Published online at Supplementry Figure S1. Ft-1 mice exhibit reduced diposity when fed n HFHS diet. WT nd ft-1 mice were fed either control or n HFHS diet for 18 weeks. A: Representtive photogrphs for side-by-side comprison

More information

* * * * * liver kidney ileum. Supplementary Fig.S1

* * * * * liver kidney ileum. Supplementary Fig.S1 Supplementry Fig.S1 liver kidney ileum Fig.S1. Orlly delivered Fexrmine is intestinlly-restricted Mice received vehicle or Fexrmine (100mg/kg) vi per os (PO) or intrperitonel (IP) injection for 5 dys (n=3/group).

More information

ESM Table 1. Characterisation of the human non-diabetic cohort used for MRIbased assessment of pancreatic fat and insulin secretion via OGTT.

ESM Table 1. Characterisation of the human non-diabetic cohort used for MRIbased assessment of pancreatic fat and insulin secretion via OGTT. ESM Tle 1. Chrcteristion of the humn non-dietic cohort used for MRIsed ssessment of pncretic ft nd insulin secretion vi OGTT. Trit sex Medin (IQR) 86 femles, 5 mles ge (yers) 4.4 (.5-5.57) BMI (kg/m²).62

More information

Supplementary Figure 1

Supplementary Figure 1 doi: 1.138/nture6188 SUPPLEMENTARY INFORMATION Supplementry Figure 1 c CFU-F colonies per 1 5 stroml cells 14 12 1 8 6 4 2 Mtrigel plug Neg. MCF7/Rs MDA-MB-231 * * MCF7/Rs-Lung MDA-MB-231-Lung MCF7/Rs-Kidney

More information

Bioactive milk components to secure growth and gut development in preterm pigs ESTER ARÉVALO SUREDA PIGUTNET FA1401 STSM

Bioactive milk components to secure growth and gut development in preterm pigs ESTER ARÉVALO SUREDA PIGUTNET FA1401 STSM Bioctive milk components to secure growth nd gut development in preterm pigs ESTER ARÉVALO SUREDA PIGUTNET FA1401 STSM STSM Pigutnet FA1401 STSM 03/Septemer 30/Novemer/2017 (3 months) Host: Home: Thoms

More information

Feeding state and age dependent changes in melaninconcentrating hormone expression in the hypothalamus of broiler chickens

Feeding state and age dependent changes in melaninconcentrating hormone expression in the hypothalamus of broiler chickens Supplementry Mterils Epub: No 2017_23 Vol. 65, 2018 https://doi.org/10.183/bp.2017_23 Regulr pper Feeding stte nd ge dependent chnges in melninconcentrting hormone expression in the hypothlmus of broiler

More information

Acute and gradual increases in BDNF concentration elicit distinct signaling and functions in neurons

Acute and gradual increases in BDNF concentration elicit distinct signaling and functions in neurons nd grdul increses in BDNF concentrtion elicit distinct signling nd functions in neurons Yunyun Ji,, Yun Lu, Feng Yng, Wnhu Shen, Tin Tze-Tsng Tng,, Linyin Feng, Shumin Dun, nd Bi Lu,.. - Grdul (normlized

More information

Effects of physical exercise on working memory and prefrontal cortex function in post-stroke patients

Effects of physical exercise on working memory and prefrontal cortex function in post-stroke patients Effects of physicl exercise on working memory nd prefrontl cortex function in post-stroke ptients M Moriy, C Aoki, K Sktni Grdute School of Helth Sciences Reserch, Mjor of Physicl Therpy, TeikyoHeisei

More information

*** *** *** *** T-cells T-ALL. T-cells T-ALL. T-cells T-ALL. T-cells T-ALL. T-cells T-ALL. Relative ATP content. Relative ATP content RLU RLU

*** *** *** *** T-cells T-ALL. T-cells T-ALL. T-cells T-ALL. T-cells T-ALL. T-cells T-ALL. Relative ATP content. Relative ATP content RLU RLU RLU Events 1 1 1 Luciferin (μm) T-cells T-ALL 1 1 Time (min) T-cells T-ALL 1 1 1 1 DCF-DA Reltive ATP content....1.1.. T-cells T-ALL RLU 1 1 T-cells T-ALL Luciferin (μm) 1 1 Time (min) c d Control e DCFH-DA

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION X p -lu c ct ivi ty doi:.8/nture8 S CsA - THA + DAPI Merge FSK THA TUN Supplementry Figure : A. Ad-Xp luc ctivity in primry heptocytes exposed to FSK, THA, or TUN s indicted. Luciferse ctivity normlized

More information

The effect of dietary α-linolenic acid levels on regulation of omega-3 lipid synthesis in rat

The effect of dietary α-linolenic acid levels on regulation of omega-3 lipid synthesis in rat The effect of dietry α-linolenic cid levels on regultion of omeg-3 lipid synthesis in rt Wei-Chun Tu School of Agriculture Food nd Wine The University of Adelide Conversion of PUFA to LCPUFA PUFA LCPUFA

More information

Expression of Three Cell Cycle Inhibitors during Development of Adipose Tissue

Expression of Three Cell Cycle Inhibitors during Development of Adipose Tissue Expression of Three Cell Cycle Inhiitors during Development of Adipose Tissue Jiin Zhng Deprtment of Animl Sciences Advisor: Michel E. Dvis Co-dvisor: Kichoon Lee Development of niml dipose tissue Hypertrophy

More information

Effect of Aqueous Extract of Carica papaya Dry Root Powder on Lactation of Albino Rats

Effect of Aqueous Extract of Carica papaya Dry Root Powder on Lactation of Albino Rats Effect of Aqueous Extrct of Cric ppy Dry Root Powder on Lcttion of Alino Rts G. Tosswnchuntr nd S. Aritjt Deprtment of Biology Fculty of Science Ching Mi University Ching Mi 50200 Thilnd Keywords: mmmry

More information

Supplementary Figure S1

Supplementary Figure S1 Supplementry Figure S Connexin4 TroponinI Merge Plsm memrne Met Intrcellulr Met Supplementry Figure S H9c rt crdiomyolsts cell line. () Immunofluorescence of crdic mrkers: Connexin4 (green) nd TroponinI

More information

Irs-2 coordinates Igf-1 receptor-mediated β-cell development and peripheral insulin signalling

Irs-2 coordinates Igf-1 receptor-mediated β-cell development and peripheral insulin signalling Irs-2 coordintes Igf-1 receptor-medited β-cell development nd peripherl insulin signlling Dominic J. Withers 1,2 *, Deorh J. Burks 1 *, Hether H. Towery 1, Shri L. Altmuro 1, Crrie L. Flint 1 & Morris

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION . Norml Physiologicl Conditions. SIRT1 Loss-of-Function S1. Model for the role of SIRT1 in the regultion of memory nd plsticity. () Our findings suggest tht SIRT1 normlly functions in coopertion with YY1,

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi:0.08/nture0987 SUPPLEMENTARY FIGURE Structure of rbbit Xist gene. Exons re shown in boxes with romn numbers, introns in thin lines. Arrows indicte the locliztion of primers used for mplifiction. WWW.NATURE.COM/NATURE

More information

Supplementary Figure 1

Supplementary Figure 1 Roles of endoplsmic reticulum stress-medited poptosis in -polrized mcrophges during mycocteril infections Supplementry informtion Yun-Ji Lim, Min-Hee Yi, Ji-Ae Choi, Jung-hwn Lee, Ji-Ye Hn, Sung-Hee Jo,

More information

Myricetin Ameliorates Defective Post-Receptor Insulin Signaling via b-endorphin Signaling in the Skeletal Muscles of Fructose-Fed Rats

Myricetin Ameliorates Defective Post-Receptor Insulin Signaling via b-endorphin Signaling in the Skeletal Muscles of Fructose-Fed Rats ecam Advnce Access published Mrch 3, 2010 ecam 2010;Pge 1 of 9 doi:10.1093/ecm/neq017 Originl Article Ameliortes Defective Post-Receptor Insulin Signling vi b-endorphin Signling in the Skeletl Muscles

More information

Hormonal networks involved in phosphate deficiencyinduced cluster root formation of Lupinus albus L.

Hormonal networks involved in phosphate deficiencyinduced cluster root formation of Lupinus albus L. Institute of Crop Science (34h) Hormonl networks involved in phosphte deficiencyinduced cluster root formtion of Lupinus lus L. For PSP5 in Montpellier, 214 Zhengrui Wng, A.B.M. Moshiur Rhmn, Guoying Wng,

More information

TNF-α (pg/ml) IL-6 (ng/ml)

TNF-α (pg/ml) IL-6 (ng/ml) Xio, et l., Supplementry Figure 1 IL-6 (ng/ml) TNF-α (pg/ml) 16 12 8 4 1,4 1,2 1, 8 6 4 2 med Cl / Pm3CSK4 zymosn curdln Poly (I:C) LPS flgelin MALP-2 imiquimod R848 CpG TNF-α (pg/ml) IL-6 (ng/ml) 2 1.6

More information

Supplementary Materials. Viral delivery of mir-196a ameliorates the SBMA phenotype via the silencing of CELF2

Supplementary Materials. Viral delivery of mir-196a ameliorates the SBMA phenotype via the silencing of CELF2 Supplementry Mterils Virl delivery of mir-96 meliortes the SBMA phenotype vi the silencing of CELF2 Yu Miyzki, Hiroki Adchi, Mshis Ktsuno, Mkoto Minmiym, Yue-Mei Jing, Zhe Hung, Hideki Doi, Shinjiro Mtsumoto,

More information

Supplementary information for: Low bone mass and changes in the osteocyte network in mice lacking autophagy in the osteoblast lineage

Supplementary information for: Low bone mass and changes in the osteocyte network in mice lacking autophagy in the osteoblast lineage Supplementry informtion for: Low one mss nd chnges in the osteocyte network in mice lcking utophgy in the osteolst linege Mrilin Piemontese, Meld Onl, Jinhu Xiong, Li Hn, Jeff D. Thostenson, Mri Almeid,

More information

Microtubule-driven spatial arrangement of mitochondria promotes activation of the NLRP3 inflammasome

Microtubule-driven spatial arrangement of mitochondria promotes activation of the NLRP3 inflammasome Supplementry Informtion Microtuule-driven sptil rrngement of mitochondri promotes ctivtion of the NLRP3 inflmmsome Tkum Misw 1,2, Michihiro Tkhm 1,2, Ttsuy Kozki 1,2, Hnn Lee 1,2, Jin Zou 1,2, Ttsuy Sitoh

More information

ARTICLE. J. E. Bowe & A. Chander & B. Liu & S. J. Persaud & P. M. Jones

ARTICLE. J. E. Bowe & A. Chander & B. Liu & S. J. Persaud & P. M. Jones Dietologi (23) 56:783 79 DOI.7/s25-2-2828-2 ARTICLE The permissive effects of glucose on receptor-operted potentition of insulin secretion from mouse islets: role for ERK/2 ctivtion nd cytoskeletl remodelling

More information

Myricetin Ameliorates Defective Post-Receptor Insulin Signaling via

Myricetin Ameliorates Defective Post-Receptor Insulin Signaling via Evidence-Bsed Complementry nd Alterntive Medicine Volume 211, Article ID 15752, 9 pges doi:1.193/ecm/neq17 Originl Article Ameliortes Defective Post-Receptor Insulin Signling vi β-endorphin Signling in

More information

Serum nesfatin-1 levels are decreased in pregnant women newly diagnosed with gestational diabetes

Serum nesfatin-1 levels are decreased in pregnant women newly diagnosed with gestational diabetes originl rticle Serum nesftin-1 levels re decresed in pregnnt women newly dignosed with gesttionl dibetes Esr Nur Ademoglu 1, Suheyl Gorr 2, Muge Keskin 3, Ayse Crlioglu 4, Rifki Ucler 5, Husmettin Erdmr

More information

Copy Number ID2 MYCN ID2 MYCN. Copy Number MYCN DDX1 ID2 KIDINS220 MBOAT2 ID2

Copy Number ID2 MYCN ID2 MYCN. Copy Number MYCN DDX1 ID2 KIDINS220 MBOAT2 ID2 Copy Numer Copy Numer Copy Numer Copy Numer DIPG38 DIPG49 ID2 MYCN ID2 MYCN c DIPG01 d DIPG29 ID2 MYCN ID2 MYCN e STNG2 f MYCN DIPG01 Chr. 2 DIPG29 Chr. 1 MYCN DDX1 Chr. 2 ID2 KIDINS220 MBOAT2 ID2 Supplementry

More information

Diabetes mellitus secondary to pancreatic diseases (type 3c): The effect of smoking on the exocrine endocrine interactions of the pancreas

Diabetes mellitus secondary to pancreatic diseases (type 3c): The effect of smoking on the exocrine endocrine interactions of the pancreas 764062DVR0010.1177/1479164118764062Dibetes & Vsculr Disese ReserchŚliwińsk-Mossoń et l. reserch-rticle2018 Originl Article Dibetes mellitus secondry to pncretic diseses (type 3c): The effect of smoking

More information

THE EVALUATION OF DEHULLED CANOLA MEAL IN THE DIETS OF GROWING AND FINISHING PIGS

THE EVALUATION OF DEHULLED CANOLA MEAL IN THE DIETS OF GROWING AND FINISHING PIGS THE EVALUATION OF DEHULLED CANOLA MEAL IN THE DIETS OF GROWING AND FINISHING PIGS THE EVALUATION OF DEHULLED CANOLA MEAL IN THE DIETS OF GROWING AND FINISHING PIGS John F. Ptience nd Doug Gillis SUMMARY

More information

Abstract ABSTRACT #69. Abstract. Introduction & Methods. Methods & Results. Results. Results & Conclusions

Abstract ABSTRACT #69. Abstract. Introduction & Methods. Methods & Results. Results. Results & Conclusions Effects of dietry β-glucn on Growth Performnce, Dirrhe, nd Gut Permeility of Wening Pigs Experimentlly Infected with Pthogenic E. coli Kwngwook Kim, Amy Ehrlich, Vivin Perng, Jennifer Chse, Helen Ryould,

More information

DOI: 10.1038/nc2331 PCre;Ros26R 12 h induction 48 h induction Vegfr3 i EC c d ib4 24 h induction VEGFR3 e Fold chnge 1.0 0.5 P < 0.05 Vegfr3 i EC Vegfr3 Figure S1 Cre ctivtion leds to genetic deletion

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION doi:1.138/nture1188 1mM CCl 2 (min) 3 4 6 CCl 2 (mm) for 4min.1. 1 (mm) Pro- d WT GdCl 3 R-68 -/- P2x7r -/- -/- Csp1 -/- WT -/- P2x7r -/- -/- Csp1 -/- Csp1 (p2) (p17) Pro-Csp1

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION DOI: 10.1038/nc2824 Hcn4 Tx5 Mlc2 c Hcn4- ISH d Tx5- ISH e Mlc2-ISH Hcn4-ISH f e Tx5-ISH f -ISH Figure S1 Section in situ hyridistion nlysis of crescent stge mouse emryos (E7.5). () More nterior section

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi:1.138/nture1794 BR EPFs BRI1? ERECTA TMM BSKs YDA PP2A BSU1 BIN2 pbzr1/2 BZR1/2 MKK4/5/7/9 MPK3/6 SPCH Cell growth Stomtl production Supplementry Figure 1. The model of BR nd stomtl signling pthwys.

More information

NappHS. rrna. transcript abundance. NappHS relative con W+W 0.8. nicotine [µg mg -1 FM]

NappHS. rrna. transcript abundance. NappHS relative con W+W 0.8. nicotine [µg mg -1 FM] (A) W+OS 3 min 6 min con L S L S RNA loding control NppHS rrna (B) (C) 8 1 k NppHS reltive trnscript undnce 6 4.5 *** *** *** *** 3 k. + + + line 1 line (D) nicotine [µg mg -1 FM] 1..8.4. con W+W Supplementl

More information

PROVEN ANTICOCCIDIAL IN NEW FORMULATION

PROVEN ANTICOCCIDIAL IN NEW FORMULATION PROVEN ANTICOCCIDIAL IN NEW FORMULATION Coxidin 100 microgrnulte A coccidiosttic dditive for roilers, chickens rered for lying nd turkeys Contins 100 g of monensin sodium per kg Aville s homogenous grnules

More information

Interleukin-4 Restores Insulin Sensitivity in Lipid-Induced Insulin-Resistant Adipocytes

Interleukin-4 Restores Insulin Sensitivity in Lipid-Induced Insulin-Resistant Adipocytes ISSN 6-2979, Biochemistry (Moscow), 21, Vol. 3, No. 5, pp. 49-56. Pleides Pulishing, Ltd., 21. Originl Russin Text I. S. Stfeev, S. S. Michurin,3, N. V. Podkuychenko, A. V. Vorotnikov, M. Yu. Menshikov,

More information

Supplementary Figure S1

Supplementary Figure S1 Supplementry Figure S1 d MAP2 GFAP e MAP2 GFAP GFAP c f Clindin GFAP Supplementry Figure S1. Neuronl deth nd ltered strocytes in the rin of n ffected child. Neuron specific MAP2 ntiody stining in the hippocmpus

More information

Check your understanding 3

Check your understanding 3 1 Wht is the difference etween pssive trnsport nd ctive trnsport? Pssive trnsport is the movement of prticles not requiring energy. Movement of prticles in ctive trnsport uses energy. 2 A gs tp in the

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION DOI: 1.138/nc286 Figure S1 e f Medium DMSO AktVIII PP242 Rp S6K1-I Gr1 + + + + + + Strvtion + + + + + IB: Akt-pT38 IB: Akt K-pT389 K IB: Rptor Gr1 shs6k1-a shs6k1-b shs6k1-c shrictor shrptor Gr1 c IB:

More information

Effect of supplemental fat from dried distillers grains with solubles or corn oil on cow performance, IGF-1, GH, and NEFA concentrations 1

Effect of supplemental fat from dried distillers grains with solubles or corn oil on cow performance, IGF-1, GH, and NEFA concentrations 1 Effect of supplementl ft from dried distillers grins with solules or corn oil on cow performnce, IGF-1, GH, nd NEFA concentrtions 1 Aigil Brtosh 2, Cody Wright 3, Aimee Wertz-Lutz 4, nd George Perry 5

More information

Keywords ATP. GK rat. Na + /K + -ATPase. Pancreatic islet. ROS. Src

Keywords ATP. GK rat. Na + /K + -ATPase. Pancreatic islet. ROS. Src Dietologi (28) 51:1226 1235 DOI 1.17/s125-8-18-x ARTICLE ctivtion genertes rective oxygen species nd impirs metolism secretion coupling in dietic Goto Kkizki nd ouin-treted rt pncretic islets R. Kominto

More information

The Journal of Physiology

The Journal of Physiology J Physiol 595.13 (2017) pp 4379 4398 4379 Impct of perintl exposure to sucrose or high fructose corn syrup (HFCS-55) on diposity nd heptic lipid composition in rt offspring Crl R. Toop 1, Beverly S. Muhlhusler

More information

Supporting Information

Supporting Information Supporting Informtion Yun et l. 1.173/pns.1523236113 SI Mterils nd Methods Humn Sujects. All prticipnts in our study were recruited from the Center for Reproductive Medicine, Shndong Provincil Hospitl

More information

Abstract. Introduction. V.I. Lushchak 1,*, T.V. Bagnyukova 1,*, J.M. Storey 2 and K.B. Storey 2

Abstract. Introduction. V.I. Lushchak 1,*, T.V. Bagnyukova 1,*, J.M. Storey 2 and K.B. Storey 2 Brzilin Journl of Medicl nd Biologicl Reserch (2001) 34: 1055-1064 Effect of exercise on glycolytic enzymes in fish tissues ISSN 0100-879X 1055 Influence of exercise on the ctivity nd the distriution etween

More information

Glucagon-Like Peptide-1 Increases Mitochondrial Biogenesis and Function in INS-1 Rat Insulinoma Cells

Glucagon-Like Peptide-1 Increases Mitochondrial Biogenesis and Function in INS-1 Rat Insulinoma Cells Brief Report Endocrinol Met 215;3:216-22 http://dx.doi.org/1.383/enm.215.3.2.216 pissn 293-596X eissn 293-5978 Glucgon-Like Peptide-1 Increses Mitochondril Biogenesis nd Function in INS-1 Rt Insulinom

More information

Supplementary Figure S1

Supplementary Figure S1 Supplementry Figure S Tissue weights (g).... Liver Hert Brin Pncres Len mss (g) 8 6 -% +% 8 6 Len mss Len mss (g) (% ody weight) Len mss (% ody weight) c Tiilis nterior weight (g).6...... Qudriceps weight

More information

Chromium Alleviates Glucose Intolerance, Insulin Resistance, and Hepatic ER Stress in Obese Mice

Chromium Alleviates Glucose Intolerance, Insulin Resistance, and Hepatic ER Stress in Obese Mice nture pulishing group rticles intervention AND prevention Chromium Allevites Glucose Intolernce, Insulin Resistnce, nd Heptic ER Stress in Oese Mice Nir Sreejyn, Feng Dong, Mchender R. Knddi,2, Xioping

More information

British Journal of Nutrition

British Journal of Nutrition British Journl of Nutrition (215), 113, 1862 1875 q The Authors 215 doi:1.117/s7114515121x Anormlities in myo-inositol metolism ssocited with type 2 dietes in mice fed high-ft diet: enefits of dietry myo-inositol

More information

British Journal of Nutrition

British Journal of Nutrition (11), 16, 1449 1456 q The Authors 11 doi:1.117/s71145111917 Fish oil comined with SCFA synergisticlly prevent tissue ccumultion of NEFA during weight loss in oese mice Miken H. Pedersen 1,, Lotte Luritzen

More information

IGF-1 vs insulin: Respective roles in modulating sodium transport via the PI-3 kinase/sgk1 pathway in a cortical collecting duct cell line

IGF-1 vs insulin: Respective roles in modulating sodium transport via the PI-3 kinase/sgk1 pathway in a cortical collecting duct cell line originl rticle http://www.kidney-interntionl.org & 27 Interntionl Society of Nephrology IGF-1 vs insulin: Respective roles in modulting sodium trnsport vi the PI-3 kinse/sgk1 pthwy in corticl collecting

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION TM TM tip link horizontl top connectors 1 leucine-rich (21 %) otoncorin-like 1809 ntigenic peptides B D signl peptide hydrophoic segment proline/threonine-rich (79 %) Supplementry Figure 1. () The outer

More information

ARTICLE. E. Pavlova 1, N. Atanassova 1, C. McKinnell 2, R.M. Sharpe 2 1 Institute of Experimental Morphology, Pathology and Anthropology with Museum,

ARTICLE. E. Pavlova 1, N. Atanassova 1, C. McKinnell 2, R.M. Sharpe 2 1 Institute of Experimental Morphology, Pathology and Anthropology with Museum, DOI:.554/5YRTIMB..3 OPPOSITE MODELS OF EXPRESSION OF ANDROGEN RECEPTOR (AR) AND RETINOIC ACID RECEPTOR-α (RAR-α) IN THE ONSET OF MALE GERM CELL DEVELOPMENT IN HORMONALLY MANIPULATED RATS E. Pvlov, N. Atnssov,

More information

The effect of encapsulated butyric acid and zinc on performance, gut integrity and meat quality in male broiler chickens 1

The effect of encapsulated butyric acid and zinc on performance, gut integrity and meat quality in male broiler chickens 1 The effect of encpsulted utyric cid nd zinc on performnce, gut integrity nd met qulity in mle roiler chickens 1 Astrct This study evluted the impct of encpsulted utyric cid nd zinc (ButiPEARL Z) on performnce

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi:0.08/nture078 RNse VifHA VifHA βctin 6 Cell lyste IP: ntiha MG VifHA VifHA β ctin 6 7 Cell lyste IP: ntiha Supplementry Figure. Effect of RNse nd MG tretment on the Vif interction., RNse tretment does

More information

Ulk λ PPase. 32 P-Ulk1 32 P-GST-TSC2. Ulk1 GST (TSC2) : Ha-Ulk1 : AMPK. WB: Ha (Ulk1) : Glu. h CON - Glu - A.A WB: LC3 AMPK-WT AMPK-DKO

Ulk λ PPase. 32 P-Ulk1 32 P-GST-TSC2. Ulk1 GST (TSC2) : Ha-Ulk1 : AMPK. WB: Ha (Ulk1) : Glu. h CON - Glu - A.A WB: LC3 AMPK-WT AMPK-DKO DOI: 10.1038/ncb2152 C.C + - + - : Glu b Ulk1 - - + λ PPse c AMPK + - + + : ATP P-GST-TSC2 WB: Flg (Ulk1) WB Ulk1 WB: H (Ulk1) GST (TSC2) C.C d e WT K46R - + - + : H-Ulk1 : AMPK - + - + + + AMPK H-Ulk1

More information

Supplemental Materials

Supplemental Materials Supplementl Mterils Cellulose deficiency of shv3svl1 is enhnced y hyper ccumultion of exogenous sucrose vi the plsm memrne sucrose/h symporter SUC1 Trevor H. Yets, Hgit Sorek, Dvid E. Wemmer, Chris R.

More information

Cyanidin-3-O-glucoside ameliorates lipid and glucose accumulation in C57BL/6J mice via activation of PPAR-α and AMPK

Cyanidin-3-O-glucoside ameliorates lipid and glucose accumulation in C57BL/6J mice via activation of PPAR-α and AMPK 3 rd Interntionl Conference nd Exhiition on Nutrition & Food Sciences Septemer 23-25, 214 Vlenci, Spin Cynidin-3-O-glucoside meliortes lipid nd glucose ccumultion in C57BL/6J mice vi ctivtion of PPAR-α

More information

Heparanase promotes tumor infiltration and antitumor activity of CAR-redirected T- lymphocytes

Heparanase promotes tumor infiltration and antitumor activity of CAR-redirected T- lymphocytes Supporting Online Mteril for Heprnse promotes tumor infiltrtion nd ntitumor ctivity of -redirected T- lymphocytes IgnzioCrun, Brr Svoldo, VlentinHoyos, Gerrit Weer, Ho Liu, Eugene S. Kim, Michel M. Ittmnn,

More information

Effect of fungicide timing and wheat varietal resistance on Mycosphaerella graminicola and its sterol 14 α-demethylation-inhibitorresistant

Effect of fungicide timing and wheat varietal resistance on Mycosphaerella graminicola and its sterol 14 α-demethylation-inhibitorresistant Effect of fungicide timing nd whet vrietl resistnce on Mycospherell grminicol nd its sterol 14 α-demethyltion-inhiitorresistnt genotypes Didierlurent L., Roisin-Fichter C., Snssené J., Selim S. Pltform

More information

TLR7 induces anergy in human CD4 + T cells

TLR7 induces anergy in human CD4 + T cells TLR7 induces nergy in humn CD T cells Mrgrit Dominguez-Villr 1, Anne-Sophie Gutron 1, Mrine de Mrcken 1, Mrl J Keller & Dvid A Hfler 1 The recognition of microil ptterns y Toll-like receptors (TLRs) is

More information

The Effect of Substituting Sugar with Artificial. Sweeteners on the Texture and Palatability of Pancakes

The Effect of Substituting Sugar with Artificial. Sweeteners on the Texture and Palatability of Pancakes The Effect of Sustituting Sugr with Artificil NUTR 453 Sweeteners on the Texture nd Pltility of Pnckes Jmie Wldron, Rquel Reyes, nd Reecc Legi 1 I. Astrct The effects of replcing sugr with Stevi nd Splend

More information

PNEUMOVAX 23 is recommended by the CDC for all your appropriate adult patients at increased risk for pneumococcal disease 1,2 :

PNEUMOVAX 23 is recommended by the CDC for all your appropriate adult patients at increased risk for pneumococcal disease 1,2 : PNEUMOVAX 23 is recommended y the CDC for ll your pproprite dult ptients t incresed risk for pneumococcl disese 1,2 : Adults ged

More information

Common genetic variation in the melatonin receptor 1B gene (MTNR1B) is associated with decreased early-phase insulin response

Common genetic variation in the melatonin receptor 1B gene (MTNR1B) is associated with decreased early-phase insulin response Dietologi (2009) 52:1537 1542 DOI 10.1007/s00125-009-1392-x ARTICLE Common genetic vrition in the meltonin receptor 1B gene (MTNR1B) is ssocited with decresed erly-phse insulin response C. Lngenerg & L.

More information

EFFECTS OF INGREDIENT AND WHOLE DIET IRRADIATION ON NURSERY PIG PERFORMANCE

EFFECTS OF INGREDIENT AND WHOLE DIET IRRADIATION ON NURSERY PIG PERFORMANCE Swine Dy 21 EFFECTS OF INGREDIENT AND WHOLE DIET IRRADIATION ON NURSERY PIG PERFORMANCE J. M. DeRouchey, M. D. Tokch, J. L. Nelssen, R. D. Goodbnd, S. S. Dritz 1, J. C. Woodworth, M. J. Webster, B. W.

More information

A. Kinoshita 1, L. Locher 2, R. Tienken 3, U. Meyer 3, S. Dänicke 3, J. Rehage 4, K. Huber 5

A. Kinoshita 1, L. Locher 2, R. Tienken 3, U. Meyer 3, S. Dänicke 3, J. Rehage 4, K. Huber 5 Effects of dietry nicin supplementtion on heptic expression of FoxO nd genes involved in glucose production in diry cows during the trnsition period A. Kinoshit, L. Locher, R. Tienken 3, U. Meyer 3, S.

More information

Responses of skeletal muscle lipid metabolism in rat gastrocnemius to hypothyroidism and iodothyronine administration: a putative role for FAT/CD36

Responses of skeletal muscle lipid metabolism in rat gastrocnemius to hypothyroidism and iodothyronine administration: a putative role for FAT/CD36 Am J Physiol Endocrinol Met 33: E1222 E1233, 212. First pulished Septemer 11, 212; doi:1.1152/jpendo.37.212. Responses of skeletl muscle lipid metolism in rt gstrocnemius to hypothyroidism nd iodothyronine

More information

Background Pears (Pyrus L.) are one of the leading cultivated fruit trees in China following apples and oranges in planting area and fruit yield.

Background Pears (Pyrus L.) are one of the leading cultivated fruit trees in China following apples and oranges in planting area and fruit yield. Nnjing Agriculturl University Potssium enhnces the sugr ssimiltion in leves nd fruit y regulting the expression of key genes involved in sugr metolism of Asin pers Cixi Dong, Chngwei Shen, Yngchun Xu College

More information

Effects of Sini San used alone and in combination with fluoxetine on central and peripheral 5-HT levels in a rat model of depression

Effects of Sini San used alone and in combination with fluoxetine on central and peripheral 5-HT levels in a rat model of depression Online Sumissions:http://www.journltcm.com J Trdit Chin Med 2013 Octoer 15; 33(5): 674-681 info@journltcm.com ISSN 0255-2922 2013 JTCM. All rights reserved. EXPERIMENTAL STUDY TOPIC Effects of Sini Sn

More information

PDGF-BB secreted by preosteoclasts induces angiogenesis during coupling with osteogenesis

PDGF-BB secreted by preosteoclasts induces angiogenesis during coupling with osteogenesis Supplementry Informtion PDGF-BB secreted y preosteoclsts induces ngiogenesis during coupling with osteogenesis Hui Xie, Zhung Cui, Long Wng, Zhuying Xi, Yin Hu, Lingling Xin, Chngjun Li, Ling Xie, Jnet

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi:1.138/nture1228 Totl Cell Numer (cells/μl of lood) 12 1 8 6 4 2 d Peripherl Blood 2 4 7 Time (d) fter nti-cd3 i.p. + TCRβ + IL17A + cells (%) 7 6 5 4 3 2 1 Totl Cell Numer (x1 3 ) 8 7 6 5 4 3 2 1 %

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi:10.1038/nture09663 Scrmle shnlrp3 shcsp1 IL-1β (p17) IL-1β (pg/ml) 2000 1500 1000 500 Wt Nlrp3-/- Ipf-/- 0 APDC IL-1β (p17) Supplementl Figure 1. Mitochondril ROS cn trigger NLRP3 inflmmsome ctivtion,

More information

SYNOPSIS Final Abbreviated Clinical Study Report for Study CA ABBREVIATED REPORT

SYNOPSIS Final Abbreviated Clinical Study Report for Study CA ABBREVIATED REPORT Finl Arevited Clinicl Study Report Nme of Sponsor/Compny: Bristol-Myers Squi Ipilimum Individul Study Tle Referring to the Dossier (For Ntionl Authority Use Only) Nme of Finished Product: Yervoy Nme of

More information

Evidence for facilitated lactate uptake in lizard skeletal muscle

Evidence for facilitated lactate uptake in lizard skeletal muscle The Journl of Experimentl Biology 24, 499 46 (2) Printed in Gret Britin The Compny of Biologists Limited 2 JEB3537 499 Evidence for fcilitted lctte uptke in lizrd skeletl muscle E. R. Donovn* nd T. T.

More information

Meat and Food Safety. B.A. Crow, M.E. Dikeman, L.C. Hollis, R.A. Phebus, A.N. Ray, T.A. Houser, and J.P. Grobbel

Meat and Food Safety. B.A. Crow, M.E. Dikeman, L.C. Hollis, R.A. Phebus, A.N. Ray, T.A. Houser, and J.P. Grobbel Met nd Food Sfety Needle-Free Injection Enhncement of Beef Strip Loins with Phosphte nd Slt Hs Potentil to Improve Yield, Tenderness, nd Juiciness ut Hrm Texture nd Flvor B.A. Crow, M.E. Dikemn, L.C. Hollis,

More information

Enhanced Chemopreventive Effect by Combining Quercetin and Green tea in Prostate Cancer

Enhanced Chemopreventive Effect by Combining Quercetin and Green tea in Prostate Cancer Enhnced Chemopreventive Effect y Comining Quercetin nd Green te in Prostte Cncer Piwen Wng, MD, PhD Assistnt Professor, Division of Cncer Reserch nd Trining Chrles R. Drew University of Medicine nd Science

More information

Input from external experts and manufacturer on the 2 nd draft project plan Stool DNA testing for early detection of colorectal cancer

Input from external experts and manufacturer on the 2 nd draft project plan Stool DNA testing for early detection of colorectal cancer Input externl experts nd mnufcturer on the 2 nd drft project pln Stool DNA testing for erly detection of colorectl cncer (Project ID:OTJA10) All s nd uthor s replies on the 2nd drft project pln Stool DNA

More information

% Inhibition of MERS pseudovirus infection. 0 h 0.5 h 1 h 2 h 4 h 6 h Time after virus addition

% Inhibition of MERS pseudovirus infection. 0 h 0.5 h 1 h 2 h 4 h 6 h Time after virus addition % Inhiition of MERS pseudovirus infection 1 8 h.5 h 1 h 2 h 4 h 6 h Time fter virus ddition Supplementry Figure S1. Inhiition of on MERS pseudovirus infection t the different intervls postinfection. A

More information

Curcumin attenuates Nrf2 signaling defect, oxidative stress in muscle and glucose intolerance in high fat diet-fed mice

Curcumin attenuates Nrf2 signaling defect, oxidative stress in muscle and glucose intolerance in high fat diet-fed mice Online Sumissions: http://www.wjgnet.com/1948-938of fice wjd@wjgnet.com doi:239/wjd.v3.i.94 World J Dietes 212 My 1; 3(): 94-14 ISSN 1948-938 (online) 212 Bishideng. All rights reserved. ORIGINAL ARTICLE

More information

De novo lipogenesis in human fat and liver is linked to ChREBP-b and metabolic health

De novo lipogenesis in human fat and liver is linked to ChREBP-b and metabolic health Received 2 Aug 2 Accepted 23 Jn 213 Pulished 26 Fe 213 DOI: 1.13/ncomms253 De novo lipogenesis in humn ft nd liver is linked to ChREBP- nd metolic helth Leh Eissing 1, *, Thoms Scherer 2, *,w, Klus Tödter

More information

Effect of processing on in vitro bioaccessibility of phenolics, flavonoids and antioxidant activity of vegetables with/without yoghurt

Effect of processing on in vitro bioaccessibility of phenolics, flavonoids and antioxidant activity of vegetables with/without yoghurt Effect of processing on in vitro ioccessiility of phenolics, flvonoids nd ntioxidnt ctivity of vegetles with/without yoghurt Assoc. Prof. Dr. Esr ÇAPANOĞLU GÜVEN Deprtment of Food Engineering Istnul Technicl

More information

Invasive Pneumococcal Disease Quarterly Report July September 2018

Invasive Pneumococcal Disease Quarterly Report July September 2018 Invsive Pneumococcl Disese Qurterly Report July Septemer Introduction Since 17 Octoer 2008, invsive pneumococcl disese (IPD) hs een notifile to the locl Medicl Officer of Helth under the Helth Act 1956.

More information

INFLUENCE OF DIFFERENT STRAINS AND WAYS OF INOCULATION ON THE RABBIT S RESPONSE TO EXPERIMENTAL INFECTION WITH PASTEURELLA MULTOCIDA

INFLUENCE OF DIFFERENT STRAINS AND WAYS OF INOCULATION ON THE RABBIT S RESPONSE TO EXPERIMENTAL INFECTION WITH PASTEURELLA MULTOCIDA Pthology nd Hygiene INFLUENCE OF DIFFERENT STRAINS AND WAYS OF INOCULATION ON THE RABBIT S RESPONSE TO EXPERIMENTAL INFECTION WITH PASTEURELLA MULTOCIDA Kulcsár G. 1, Fáián K. 1 *, Brn T. 1, Virág Gy.

More information

Flaxseed Lignan Increased Glucose Uptake by Human Red Blood Cells

Flaxseed Lignan Increased Glucose Uptake by Human Red Blood Cells The Open Nutrceuticls Journl, 29, 2, 81-85 81 Open Access Flxseed Lignn Incresed Glucose Uptke y Humn Red Blood Cells Yeong Rhee * nd Ardith Brunt 351 EML, NDSU Dept. # 262, PO Box 65, North Dkot Stte

More information

Critical role of c-kit in beta cell function: increased insulin secretion and protection against diabetes in a mouse model

Critical role of c-kit in beta cell function: increased insulin secretion and protection against diabetes in a mouse model Dietologi (1) 55:14 5 DOI 1.17/s15-1-5-5 ARTICLE Criticl role of c-kit in et cell function: incresed insulin secretion nd protection ginst dietes in mouse model Z. C. Feng & J. Li & B. A. Turco & M. Riopel

More information

Synergistic effects of metformin, resveratrol, and hydroxymethylbutyrate on insulin sensitivity

Synergistic effects of metformin, resveratrol, and hydroxymethylbutyrate on insulin sensitivity Dietes, Metolic Syndrome nd Oesity: Trgets nd Therpy Open Access Full Text Article open ccess to scientific nd medicl reserch Originl Reserch Synergistic effects of metformin, resvertrol, nd hydroxymethylutyrte

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION doi:10.1038/nture11225 Numer of OTUs sed on 3% distnce Numer of 16s rrna-sed V2-V4 tg sequences LF MF PUFA Supplementry Figure 1. High-ft diets decrese the richness nd diversity

More information

Chapter 5: The peripheral nervous system Learning activity suggested answers

Chapter 5: The peripheral nervous system Learning activity suggested answers Chpter 5: The peripherl nervous system Lerning ctivity suggested nswers Lerning Activity 5.1 (p. 222) 1 Briefly descrie the two min functions of the somtic nervous system. Description should refer to:

More information

Electronic Supplementary Information for:

Electronic Supplementary Information for: Electronic Supplementry Mteril (ESI) for ChemComm. This journl is The Royl Society of Chemistry 214 Electronic Supplementry Informtion for: Gold nnoprticles functionlized with cresyl violet nd porphyrin

More information

Chronic high-sodium diet intake after weaning lead to neurogenic hypertension in adult Wistar rats

Chronic high-sodium diet intake after weaning lead to neurogenic hypertension in adult Wistar rats Chronic high-sodium diet intke fter wening led to neurogenic hypertension in dult Wistr rts 1 Pul Mglhães Gomes; 2 Rento Willin Mrtins Sá; 1 Giovn Lopes Aguir; 1 Milede Hnner Sriv Pes; 1 Andréi Crvlho

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi:10.1038/nture09973 Plsm Memrne Phgosome TLR1/2/4 ROS Mitochondrion ROS OXPHOS Complex I ROS TRAF6 NADPH Oxidse Supplementry Figure 1 Model detiling the roles of mitochondril ROS in mcrophge cteril

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi: 1.138/nture862 humn hr. 21q MRPL39 murine Chr.16 Mrpl39 Dyrk1A Runx1 murine Chr. 17 ZNF295 Ets2 Znf295 murine Chr. 1 COL18A1 -/- lot: nti-dscr1 IgG hevy hin DSCR1 DSCR1 expression reltive to hevy

More information

FERTILITY EFFECTS OF SODIUM FLUORIDE IN MALE MICE

FERTILITY EFFECTS OF SODIUM FLUORIDE IN MALE MICE 128 Fluoride Vol. 33 No. 3 128-134 2000 Reserch Report FERTILITY EFFECTS OF SODIUM FLUORIDE IN MALE MICE Ahmed Elbetieh, Hom Drmni, Ahmd S Al-Hiyst b Irbid, Jordn SUMMARY: Sexully mture mle Swiss mice

More information

Clinical Study Report Synopsis Drug Substance Naloxegol Study Code D3820C00018 Edition Number 1 Date 01 February 2013 EudraCT Number

Clinical Study Report Synopsis Drug Substance Naloxegol Study Code D3820C00018 Edition Number 1 Date 01 February 2013 EudraCT Number EudrCT Number 2012-001531-31 A Phse I, Rndomised, Open-lbel, 3-wy Cross-over Study in Helthy Volunteers to Demonstrte the Bioequivlence of the Nloxegol 25 mg Commercil nd Phse III Formultions nd to Assess

More information

PHYSIOLOGICAL AND PROTEOMIC RESPONSES OF TOBACCO SEEDLINGS EXPOSED TO SILVER NANOPARTICLES

PHYSIOLOGICAL AND PROTEOMIC RESPONSES OF TOBACCO SEEDLINGS EXPOSED TO SILVER NANOPARTICLES PHYSIOLOGICAL AND PROTEOMIC RESPONSES OF TOBACCO SEEDLINGS EXPOSED TO SILVER NANOPARTICLES Rent Bi Deprtment of Biology, Fculty of Science, University of Zgre INTRODUCTION Nnoprticles (NPs) Silver nnoprticles

More information