Expression of Three Cell Cycle Inhibitors during Development of Adipose Tissue

Size: px

Start display at page:

Download "Expression of Three Cell Cycle Inhibitors during Development of Adipose Tissue"

Clifford Waters
5 years ago
Views:

1 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

2 Development of niml dipose tissue Hypertrophy Hyperplsi Increse Ft cell size Ft cell numer Occur stge Mechnism Whole developmentl stge Increse deposition of lipid Bckground Normlly t erly stge, ut lso in severely oese dults Prolifertion nd differentition of predipocytes Cell cycle Oesity Medium Severe (BMI>35) 2

3 Bckground Cell cycle during dipocyte differentition In Vitro Cell Cycle Confluent Predipocyte Mitotic Clonl Expnsion Terminlly Differentited Adipocyte In Vivo Cell Cycle Predipocyte Immture dipocyte Lrge Nonprolifertive Adipocyte Smll prolifertive dipocyte (SPA) 3

4 Hypothesis Core inhiitors in cell cycle regultion of ft cells Tissue Distriution Predipocytes (P) vs. Adipocytes (A) Adipocyte differentition in vitro Development of dipose tissue in vivo Expressed highly in mture dipose tissue P<A 4

5 GDS596 (Humn, n=2) GEO Dt Anlysis Dtmining in gene expression omnius (GEO) dtsets (GDS) GDS3142 (Mouse,n=3 or 4) Six tissues (dipose, hert, liver, lung, muscle, kidney) dipose verge of other five Descending rnk Multiple comprisons with MIXED model in SAS Cell cycle inhiitors with high rnks 5

6 GEO Dt Anlysis Selection of cell cycle inhiitors sed on GDS596 (Humn) nd GDS3142 (Mouse) Gene (Rnk) Adipose Muscle Hert Lung Liver Kidney A/O CCNG2(2102) c 138 c 45 c Humn CDKN2C(654) c 310 c 92 c 42 c PMP22 (2023) c e 369 d 1.43 CCNG2(1878) c c 399 c 1.69 Mouse CDKN2C (55) c c 253 c 9.55 PMP22(2007) c 668 c e 248 d 1.65 A/O: rtio etween expression in dipose tissue nd verge expression in other tissues. Different letters (-e) : significnt difference (P<5). Cyclin-G2(CCNG2), Cyclin-dependent kinse inhiitor (CDKN2C), Peripherl myelin protein 22 (PMP22 ) 6

7 GEO Dt Anlysis Selected cell cycle inhiitors Cyclin-G2 (CCNG2): Unconventionl cyclin; inhiits cell cycle t G1/S trnsition (Aguilr et l., 2010). Cyclin-dependent kinse inhiitor 2C (CDKN2C): Key cell cycle inhiitor, inds to Cyclin dependent kinse 4 nd 6 (CDK4 nd CDK6) in G1 phse (Hiri et l., 1995). Peripherl myelin protein 22 (PMP22): growth-rrest specific gene 3 (gs3), mintins G0 growth rrest (Welcher et l., 1991). 7

8 Moleculr Reserch qpcr to detect expression in porcine dipose tissues 105 dy fetl Lndrce 0 Dy 3 Dy 6 Dy 9 Dy 6 dy postntl Lndrce In Vivo Tissue Development In Vitro Cell Culture Sucutneous Adipose (n=3) Stroml Vsculr (SV) 120 dy mrketge Lndrce vs. Ft Cell (FC) Internl Control: Cyclophilin (Cyc) Tissue Distriution Adipose Hert Liver Lung Muscle Kidney Spleen Intestine 8

9 GEO Dt Anlysis Dt processing nd nlysis Multiple comprison of the selected genes: MIXED model with DIFF option in SAS. Pirwise comprison etween undifferentited nd differentiting/ed predipocytes: Student s t test in SAS. 9

10 Trget gene/cyc Moleculr Reserch Expression of the three cell cycle inhiitors in vrious tissues in pigs CCNG2 CDKN2C PMP22 cd c d cd cd c F M H Lu Li K S I F M H Lu Li K S I F M H Lu Li K S I Tissues: Ft(F), Muscle(M), Hert(H), Lung(Lu), Liver(Li), Kidney (K), Spleen(S) nd Intestine(I). Different letters (-c) indicte significnt difference (P<5). c High expression occurs in muscle or lung, low expression occurs in liver. Expression in dipose tissue is close to men expression in different tissues. c c c c c cde e e de cd 10

11 Trget gene/cyc Moleculr Reserch Expression of the three genes in stroml vsculr cell (SVC) vs. ft cell (FC) in pigs DLK1 PPARγ CCNG2 CDKN2C PMP * *** ** ** FC SVC SVC FC *: P<5; **: P<1 ***:P: <01 SVC<FC for ll three genes. DLK1- predipocyte mrker is high in SVC; PPARγ n dipocyte mrker - is high in FC. Seprtion of SVC nd FC ws successful. 11

12 Trget gene/cyc Moleculr Reserch Expression during in vitro differentition of stroml-vsculr cells DLK1 PPARγ CCNG2 CDKN2C PMP c c Different letters (-c) indicte significnt difference (P<5) c c c c 4.0 Dy 0 Dy 3 Dy 6 Dy Three cell cycle inhiitors nd PPARγ incresed during cell culture. DLK1 dropped to low level fter differentition. 12

13 Trget gene/cyc Moleculr Reserch Expression during dipose development DLK1 PPARγ CCNG2 CDKN2C PMP Different letters (-c) indicte significnt difference (P<5) Emryo 6 d old Adult Emryo: High expression of DLK1 Predipocytes is dominnt 6-dy pigs: Low expression of ll the genes SPA is dominnt 120-dy pigs: High expression of three cell cycle inhiitors nd PPAR γ Aipocytes is dominnt

14 Conclusion Tissue Distriution Predipocytes vs. Adipocytes Adipocyte differentition in vitro Development of dipose tissue in vivo Core inhiitors in cell cycle regultion of ft cells Expressed highly in dipose tissue P<A Conclusion Age or species? 14

15 Conclusion CCNG2, CDKN2C nd PMP22 re involved in: Cell cycle inhiition during ft cell differentition Mintin quiescent stte for hypertrophy Regultion during development of dipose tissue Close reltionship etween the three genes nd development of dipose tissue Possile mrkers for selection of less ft deposition in niml industry Potentil wys to prevent severe oesity in humn SPAs my e dominnt in postntl pigs 15

16 Future Pln Western Blotting: Confirm results in protein level Proliferting cellulr nucler ntigen (PCNA) Immunostining: Confirm cell prolifertion sttus during in vivo development of dipose tissue Identifiction of core cell cycle ctivtors during hyperplsi 16

17 Acknowledgement The Ohio Agriculturl Reserch nd Development Center, The Ohio Stte University. The Agend Progrm (No. PJ008587), Rurl Development Administrtion, Repulic of Kore. Dr. Michel E. Dvis Dr. Kichoon Lee Dr. Jinsoo Ahn Dr. Young M. Choi Yeunsu Suh 17

18 18

Differential expression of cyclin G2, cyclin dependent kinase inhibitor 2C and peripheral myelin protein 22 genes during adipogenesis..

Differential expression of cyclin G2, cyclin dependent kinase inhibitor 2C and peripheral myelin protein 22 genes during adipogenesis.. The Ohio Stte University From the SeletedWorks of Jiin Zhng Summer My 5, 214 Differentil expression of ylin G2, ylin dependent kinse inhiitor 2C nd peripherl myelin protein 22 genes during dipogenesis..pdf