1 Molecular cloning and characterization of KISS1 promoter and effect of KISS1 gene mutations on litter size in the goat X.P. An, P. Han, J.X. Hou, H.B. Zhao, Y. Yan, T. Ma, F. Fang, F.X. Meng, Y.X. Song, J.G. Wang and B.Y. Cao College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China Corresponding author: B.Y. Cao Genet. Mol. Res. 12 (4): (2013) Received May 29, 2012 Accepted November 10, 2012 Published February 28, 2013 DOI ABSTRACT. Kisspeptins, the product of the KISS1 gene, play an essential role in the regulation of reproductive functions, acting primarily at the hypothalamic level of the gonadotropic axis. We detected polymorphisms of the goat KISS1 gene in 723 individuals from three goat breeds (Xinong Saanen, Guanzhong, and Boer) by DNA pooling, PCR-RFLP, and DNA sequencing methods. We cloned the promoter sequence of this gene and found it to share high similarity with that of the bovine KISS1 promoter. Six TATA boxes were found in the goat KISS1 promoter region. Two novel SNPs (g.2124t>a and g.2270c>t) were identified in the intron 1 of the KISS1 gene of all three goat breeds. The three goat breeds were in Hardy-Weinberg disequilibrium at g.2124t>a and g.2270c>t loci. The g.2124t>a and g.2270c>t loci were closely linked in the three goat breeds (r 2 > 0.33). The g.2124t>a and g.2270c>t SNPs were significantly associated with litter size, and the C1 female goats had a larger litter size than did those with the other genotypes. These results extend the spectrum
2 Effect of KISS1 gene mutations on goat litter size 4309 of genetic variation of the goat KISS1 gene, which contributes to our knowledge of goat genetic resources for breeding programs. Key words: Combined genotype; SNP; PCR-RFLP; Goat INTRODUCTION In livestock, litter size is the result of well-regulated interactions of endocrine and paracrine mediators. Genetic information, particularly for those loci that affect performance traits, is an important tool in breeding programs. Extensive research has been carried out on different prolific goat breeds to identify the genes involved in the control of litter size (Polley et al., 2009). Several candidate genes and their relationship with litter size in goats have been studied, such as the G protein-coupled receptor 54 (GPR54) gene (Cao et al., 2011), cocaine-amphetamine-regulated transcript (CART) gene (Wang et al., 2011), kit ligand (KITLG) gene (An et al., 2012), bone morphogenetic protein receptor IB (BMPR-IB) gene (Chu et al., 2010) and so on. The KISS1 gene encodes a family of neuropeptides called kisspeptins, which activate receptor G protein-coupled receptor-54 and play a role in the neuroendocrine regulation of GnRH secretion (Smith et al., 2005). Moreover, KISS1 neurons in the hypothalamus participate in crucial features of reproductive maturation and function, such as brain-level sex differentiation, puberty onset and the neuroendocrine regulation of gonadotropin secretion and ovulation (Caraty et al., 2010). Kisspeptins are very potent elicitors of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion in different mammalian species (Gottsch et al., 2004; Dhillo et al., 2005). Specifically in the female, the hypothalamic KISS1/ GPR54 system seems to operate as a central conduit for not only the negative but also the positive feedback regulation of gonadotropins, thereby playing a substantial role in the generation of the preovulatory surge of LH (Kinoshita et al., 2005; Smith et al., 2006). In sheep, KISS1 mrna-expressing cells are found in the arcuate nucleus (ARC) and dorsallateral preoptic area, and both appear to mediate the positive feedback effect of estradiol to generate the preovulatory GnRH/LH surge (Smith et al., 2011). In addition to their prominent expression at hypothalamic levels, fragmentary evidence suggests that KISS1 and or KISS1R mrnas or proteins are also present in several peripheral reproductive tissues including the ovarie (de Roux et al., 2003), oviducts (Gaytan et al., 2007) and testes (Ohtaki et al., 2001). In good agreement, kisspeptin immunoreactivity has been observed in cycling human and marmoset ovaries, with prominent signals in the theca layer of growing follicles, corpora lutea, interstitial gland and ovarian surface epithelium (Gaytan et al., 2009). KISS1 knockout mice are viable and healthy with no apparent abnormalities but fail to undergo sexual maturation. Mutant female mice do not progress through the estrous cycle (d Anglemont de Tassigny et al., 2007). One novel nonsynonymous single nucleotide polymorphism (G T) substituting one amino acid in kisspeptin (P110T) has been found to be statistically related to central precocious puberty in human (Luan et al., 2007). These findings indicate that the KISS1 gene could be an excellent candidate gene for reproductive traits in humans and livestock. In view of the above considerations, the objectives of the present study were to determine the polymorphisms of the caprine KISS1 gene in three breeds and to investigate the associations between these genetic markers and litter size.
3 X.P. An et al MATERIAL AND METHODS Genomic DNA isolation and data collections Blood samples were obtained from 723 goats belonging to three breeds: Xinong Saanen (SN; N = 306), Guanzhong (GZ; N = 221) and Boer (BG; N = 196). They were reared in Qianyang, Zhouzhi and Liuyou counties of Shaanxi Province, respectively. All diets were based on alfalfa, corn silage, and a combination of concentrates including corn, soybean meal, and bone meal. Health, fertility and production records were maintained by the dairymen and veterinarians. The litter size from the first to fourth parity was obtained from production records. Five milliliters blood per goat were collected aseptically from the jugular vein and kept in a tube containing ACD anticoagulant (citric acid, sodium citrate and dextrose - 10:27:38). All samples were delivered to the laboratory in an ice box. The genomic DNA was extracted from white blood cells using a standard phenol-chloroform extraction protocol. All experiments were performed in accordance with the National Institute of Health Guide for the Care and Use of Laboratory Animals. Cloning and sequencing of KISS1 promoter Using the Primer software (version 6.0) and according to the bovine KISS1 gene (GenBank accession No. NW_ ), one pair of primers was designed to amplify the goat KISS1 promoter (Table 1). The PCR products were separated on a 1.5% agarose gel, purified with the Gel Extraction kit (Tiangen, Beijing, China) and inserted into pmd19-t vector (TaKaRa, Dalian, China), according to provided protocols. The recombinant plasmid was then transformed into competent Escherichia coli JM109 cells. At least three positive clones were sequenced in both directions by Beijing Invitrogen Biotechnology Co. Ltd. (Beijing, China). Sequence analysis was performed with on-line BLAST, it/~webgene/wwwhc_tata.html and Table 1. Primer sequences for the goat KISS1 gene applied for screening polymorphisms and genotyping. Primer Sequence (5ꞌ 3ꞌ) Gene region Amplicon (bp) GenBank accession No. Tm ( C) KISS1-F1 TTCACCTGGCTGACTTGT Promoter 2069 NW_ KISS1-R1 ATACCTGTGGTTCTAGGATTC KISS1-F2 TGCAAAGCCGAGTGTGCAGG Exon GU KISS1-R2 TGAAGGCGGTGGCACAAAGG KISS1-F3 CCCGCTGTAACTAGAGAAAG Intron GU KISS1-R3 CATCCAGGGTGAGTGATACT Tm = melting temperature. SNP investigation and genotyping On the basis of the caprine KISS1 gene sequence (GenBank accession No. GU142847), two pairs of primers were designed to amplify the exon 1 and partial intron 1 of the KISS1 gene (Table 1). Their optimal annealing temperatures are shown in Table 1. Herein, we screened them to identify SNPs of this gene by the pooled DNA sequencing method (Bansal et al., 2002). Five microliters 100 ng/μl DNA per sample were collected to create a DNA pool for each goat breed.
4 Effect of KISS1 gene mutations on goat litter size 4311 PCR products were sent to Beijing Genomics Institute (Beijing, China) to be sequenced in both directions. SNPs were detected using Chromas 2.31 and DNAstar 7.0 softwares. The SNPs of the KISS1 gene were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The 25-μL reaction volume contained 50 ng genomic DNA, 12.5 µl 2X reaction mix (including 500 µm dntps, 20 mm Tris-HCl, ph 9, 100 mm KCl, 3 mm MgCl 2 ), 0.5 µm of each primer, and 0.5 U Taq DNA polymerase. The cycling protocol was 5 min at 95 C and 35 cycles of denaturation at 94 C for 30 s, annealing at X C (Table 1) for 30 s, and extension at 72 C for 35 s, with a final extension at 72 C for 10 min. PCR products (5 μl) of different primer pairs were mixed with 0.7 μl 10X buffer, 2.5 U restriction enzyme (NEB, Ipswich, UK) and 3.8 μl sterile ddh 2 O, and then incubated for 1.5 h at 37 C. The restriction enzymes used are shown in Table 2. Digestion products were subjected to 3.5% horizontal agarose gel electrophoresis. The agarose gels were stained with ethidium bromide, and the genotypes were then determined. Statistical analysis Allelic frequencies, heterozygosity (H E ) and polymorphism information content (PIC) were calculated using Popgene (version 4.0). Linkage disequilibrium (LD) was assessed by the SHEsis software (Shi and He, 2005). Statistical analysis was performed using the general linear model procedure of the SPSS 16 statistical software. The model applied was: Y iklm = µ + C i + B k + (BC) ik + S l + E iklm, where Y iklm is the trait measured on each of the iklm th animal, µ is the overall population mean, C i is the fixed effect associated with the i th combined genotype, B k is the fixed effect associated with the k th breed, (BC) ik is the interaction between the i th combined genotype and the k th breed, S l is the fixed effect associated with the l th sire, and E iklm is the random error. RESULTS SNP identification and genotypes The 2069-bp fragment, including a 1962-bp fragment upstream from the start codon and a 107-bp fragment of exon 1 in the goat KISS1 gene, was amplified (Figure 1). The sequence of the KISS1 promoter was submitted to the National Center for Biotechnology Information (GenBank accession No. JX047312). A comparison with the bovine KISS1 5ꞌ-flanking sequence demonstrated a high degree of homology with approximately 88.59% nucleotide identity. The transcription factor binding sites of the KISS1 promoter region are shown in Figure S1. Six TATA boxes were found in the KISS1 promoter region (TCTATCACTG, CTGATACCAT, GGTCTATAGC, TCTATAGCTC, ATTATAGGCA, and ACTATATGGC) (Figure S1). Two SNPs (g.2124t>a and g.2270c>t) were genotyped in the three goat breeds (Figures 2 and 3). The g.2124t>a and g.2270c>t mutations were in intron 1 (GenBank accession No. JQ806381). At g.2124t>a and g.2270c>t loci, PIC was in the three goat breeds. According to the PIC classification (low polymorphism if PIC value < 0.25, moderate polymorphism if 0.25 < PIC value < 0.50, and high polymorphism if PIC > 0.50), the goat breeds had moderate genetic diversity at g.2124t>a and g.2270c>t loci. Genotypic and allelic frequencies of the two SNPs are shown in Table 2. The SN and GZ breeds were in Hardy-Weinberg disequilibrium at the g.2124t>a and g.2270c>t loci (P < 0.05) (Table 2).
5 X.P. An et al LD was estimated in these breeds to determine the linkage relationships between the two SNPs (Table 2). If r 2 > 0.33, LD was considered strong (Ardlie et al., 2002). According to the results, both g.2124t>a and g.2270c>t loci were closely linked in the three goat breeds (Table 2). Figure 1. PCR product of the promoter region of the goat KISS1 gene. Figure 2. Electrophoresis patterns obtained after digestion with XmnI endonuclease at the g.2124t>a locus. Lane M = DNA marker; lanes 1 and 2 = TA genotype; lanes 3 and 6 = TT genotype; lanes 4 and 5 = AA genotype. Figure 3. Electrophoresis patterns obtained after digestion with MwoI endonuclease at the g.2270c>t locus. Lane M = DNA marker; lanes 1 and 4 = CC genotype; lanes 2 and 5 = TT genotype; lanes 3 and 6 = TC genotype.
6 Effect of KISS1 gene mutations on goat litter size 4313 Table 2. Genotypic distribution and allelic frequencies of two SNP loci in the KISS1 gene. Locus Restriction enzyme Breed SN GZ BG g.2124t>a XmnI Genotype TT TA AA Allele T A H E PIC Equilibrium χ 2 test P = 0.03 P = 0.03 P = 0.02 g.2270c>t MwoI Genotype TT TC CC Allele T C H E PIC Equilibrium χ 2 test P = 0.01 P = 0.01 P = 0.55 LD of g.2124t>a and g.2270c>t r a 0.71 a 0.52 a SN = Xinong Saanen; GZ = Guanzhong; BG = Boer; LD = linkage disequilibrium; H E = expected heterozygosity; PIC = polymorphism information content. a Strong linkage disequilibrium (r 2 > 0.33). Association and effects of combined genotypes The associations of 7 combined genotypes of 2 loci with litter size are shown in Table 3. In the first parity, the C1 female goats had greater litter size than did the C4, C6 or C7 goats (P < 0.05). In the second and third parity, the C1 female goats had larger litter size than did the C7 goats (P < 0.05). In the average parity, the C7 female goats had the smallest litter size compared to the C1 and C4 goats (P < 0.05), and the C1 female goats had larger litter size compared to the C4 goats (P < 0.05). Table 3. Combined effect of two SNP loci on litter size (means ± standard errors) in SN, GZ and BG breeds. Genotype N 1st parity 2nd parity 3rd parity 4th parity Average litter size litter size litter size litter size litter size C1 (TTTT) ± 0.05 b 1.85 ± 0.05 b 1.98 ± 0.05 b 2.08 ± ± 0.03 b C2 (TATT) ± ± ± ± ± 0.06 C3 (TTTC) ± ± ± ± ± 0.07 C4 (TATC) ± 0.05 a 1.73 ± ± 0.05 b 2.02 ± ± 0.03 c C5 (AATC) ± ± ± ± ± 0.09 C6 (TACC) ± 0.12 a 1.61 ± ± ± ± 0.07 C7 (AACC) ± 0.06 a 1.60 ± 0.06 a 1.70 ± 0.07 a 1.97 ± ± 0.04 a Values with different superscripts in the same column differ significantly at P < For breed abbreviations, see legend to Table 2. DISCUSSION CdxA, CEPB, GATA-1, Sp1, and MZF1 play an important role in regulating gene expression (Grange et al., 1991; Merika and Orkin, 1993; Cram et al., 2001; Liu et al., 2011), which can also be observed in the goat KISS1 promoter. In addition, TATA boxes (ATTATAGGCA and ACTATATGGC) were found in the goat and bovine KISS1 promoters.
7 X.P. An et al Therefore, they may play a similar regulatory role in both cattle and goats. Both g.2124t>a and g.2270c>t loci were closely linked in the three goat breeds studied, which may be a result of selection. Selection during domestication and improvement can influence the LD level of a gene (Saunders et al., 2005), and selection aimed at alleles of a structural gene can significantly increase the LD level in the target gene region (Clark et al., 2004). The two SNP loci were in Hardy-Weinberg disequilibrium in the SN and GZ breeds (P < 0.05), which showed that the genotypic frequencies had been affected by selection, mutation or migration. So far, there have been some studies of the KISS1 gene as a candidate gene for reproductive traits in animals, which revealed that the KISS1 gene plays an important role in animal reproduction (Tomikawa et al., 2010). Huijbregts et al. (2012) detected three SNPs (c.638inst, c.641c>g and c.645g>ca) in the 3ꞌ-UTR of the human KISS1 gene, and found that the c.645g>ca mutation was associated with central precocious puberty. Cao et al. (2010) showed an association between allele C of the 296 locus and allele deletion of the locus in the KISS1 gene and larger litter size in Jining Grey goats. Hou et al. (2011) identified T2643C and 8-bp base deletions (2677AGTTCCCC) in intron 2 of the goat KISS1 gene, with T2643C showing significant effects on litter size (P < 0.05). Reproductive traits are complex quantitative traits involving multiple genes, loci and interactions, so it is important to analyze the combined effect of multiple genes or loci on reproductive traits. In the present study, the association between multiple loci and litter size from the first to the fourth parity was analyzed. The C1 female goats had larger litter size compared to those with other combined genotypes in average parity. Although two novel variants of the KISS1 gene do not concern the coding region, it is possible that they regulate the expression of the KISS1 gene. Sequences in the non-coding region can affect the mechanism of mrna deadenylation and degradation (Xu et al., 1997; Nackley et al., 2006). Accumulating evidence further shows that central or peripheral administration of kisspeptin stimulates GnRH-dependent LH and FSH secretion in various mammalian species from rodents to humans (Gottsch et al., 2004; Navarro et al., 2005), suggesting that kisspeptin plays an essential role in governing reproductive functions throughout species. Biochemical and physiological functions, together with the results obtained in our study, indicate that the KISS1 gene could be used as a molecular breeding marker in goats. In conclusion, this study indicated that two SNPs may play an important role in litter size. The C1 female goats had a larger litter size than did those with other combined genotypes in average parity and could be used for the development of new breeds of prolific goats. ACKNOWLEDGMENTS Research supported by the National Support Program of China (#2011BAD28B05-3) and the Science and Technology Innovation Project of Shaanxi Province (#2011KTCL02-09). Supplementary material REFERENCES An XP, Hou JX, Li G, Song YX, et al. (2012). Polymorphism identification in the goat KITLG gene and association analysis with litter size. Anim. Genet. 43: Ardlie KG, Kruglyak L and Seielstad M (2002). Patterns of linkage disequilibrium in the human genome. Nat. Rev. Genet. 3:
8 Effect of KISS1 gene mutations on goat litter size 4315 Bansal A, van den Boom D, Kammerer S, Honisch C, et al. (2002). Association testing by DNA pooling: an effective initial screen. Proc. Natl. Acad. Sci. U. S. A. 99: Cao GL, Chu MX, Fang L, Di R, et al. (2010). Analysis on DNA sequence of KiSS-1 gene and its association with litter size in goats. Mol. Biol. Rep. 37: Cao GL, Chu MX, Fang L, Feng T, et al. (2011). Analysis on DNA sequence of GPR54 gene and its association with litter size in goats. Mol. Biol. Rep. 38: Caraty A, Franceschini I and Hoffman GE (2010). Kisspeptin and the preovulatory gonadotrophin-releasing hormone/ luteinising hormone surge in the ewe: basic aspects and potential applications in the control of ovulation. J. Neuroendocrinol. 22: Chu MX, Zhao XH, Zhang YJ, Jin M, et al. (2010). Polymorphisms of BMPR-IB gene and their relationship with litter size in goats. Mol. Biol. Rep. 37: Clark RM, Linton E, Messing J and Doebley JF (2004). Pattern of diversity in the genomic region near the maize domestication gene tb1. Proc. Natl. Acad. Sci. U. S. A. 101: Cram EJ, Liu BD, Bjeldanes LF and Firestone GL (2001). Indole-3-carbinol inhibits CDK6 expression in human MCF-7 breast cancer cells by disrupting Sp1 transcription factor interactions with a composite element in the CDK6 gene promoter. J. Biol. Chem. 276: d Anglemont de Tassigny, X, Fagg LA, Dixon JP, Day K, et al. (2007). Hypogonadotropic hypogonadism in mice lacking a functional Kiss1 gene. Proc. Natl. Acad. Sci. U. S. A. 104: de Roux N, Genin E, Carel JC, Matsuda F, et al. (2003). Hypogonadotropic hypogonadism due to loss of function of the KiSS1-derived peptide receptor GPR54. Proc. Natl. Acad. Sci. U. S. A. 100: Dhillo WS, Chaudhri OB, Patterson M, Thompson EL, et al. (2005). Kisspeptin-54 stimulates the hypothalamic-pituitary gonadal axis in human males. J. Clin. Endocrinol. Metab. 90: Gaytan F, Gaytan M, Castellano JM, Romero M, et al. (2009). KiSS-1 in the mammalian ovary: distribution of kisspeptin in human and marmoset and alterations in KiSS-1 mrna levels in a rat model of ovulatory dysfunction. Am. J. Physiol. Endocrinol. Metab. 296: E520-E531. Gaytan M, Castellano JM, Roa J, Sanchez-Criado JE, et al. (2007). Expression of KiSS-1 in rat oviduct: possible involvement in prevention of ectopic implantation? Cell Tissue Res. 329: Gottsch ML, Cunningham MJ, Smith JT, Popa SM, et al. (2004). A role for kisspeptins in the regulation of gonadotropin secretion in the mouse. Endocrinology 145: Grange T, Roux J, Rigaud G and Pictet R (1991). Cell-type specific activity of two glucocorticoid responsive units of rat tyrosine aminotransferase gene is associated with multiple binding sites for C/EBP and a novel liver-specific nuclear factor. Nucleic Acids Res. 19: Hou JX, An XP, Wang JG and Song YX (2011). New genetic polymorphisms of KiSS-1 gene and their association with litter size in goats. Small Rumin. Res. 96: Huijbregts L, Roze C, Bonafe G, Houang M, et al. (2012). DNA polymorphisms of the KiSS1 3 untranslated region interfere with the folding of a G-rich sequence into G-quadruplex. Mol. Cell Endocrinol. 351: Kinoshita M, Tsukamura H, Adachi S, Matsui H, et al. (2005). Involvement of central metastin in the regulation of preovulatory luteinizing hormone surge and estrous cyclicity in female rats. Endocrinology 146: Liu YF, Zan LS, Cui WT, Xin YP, et al. (2011). Molecular cloning, characterization and association analysis of the promoter region of the bovine CDK6 gene. Genet. Mol. Res. 10: Luan X, Zhou Y, Wang W, Yu H, et al. (2007). Association study of the polymorphisms in the KISS1 gene with central precocious puberty in Chinese girls. Eur. J. Endocrinol. 157: Merika M and Orkin SH (1993). DNA-binding specificity of GATA family transcription factors. Mol. Cell Biol. 13: Nackley AG, Shabalina SA, Tchivileva IE, Satterfield K, et al. (2006). Human catechol-o-methyltransferase haplotypes modulate protein expression by altering mrna secondary structure. Science 314: Navarro VM, Castellano JM, Fernandez-Fernandez R, Tovar S, et al. (2005). Characterization of the potent luteinizing hormone-releasing activity of KiSS-1 peptide, the natural ligand of GPR54. Endocrinology 146: Ohtaki T, Shintani Y, Honda S, Matsumoto H, et al. (2001). Metastasis suppressor gene KiSS-1 encodes peptide ligand of a G-protein-coupled receptor. Nature 411: Polley S, De Sachinandan, Batabyal S, Kaushik R, et al. (2009). Polymorphism of fecundity genes (BMPR1B, BMP15 and GDF9) in the Indian prolific Black Bengal goat. Small Rumin. Res. 85: Saunders MA, Slatkin M, Garner C, Hammer MF, et al. (2005). The extent of linkage disequilibrium caused by selection on G6PD in humans. Genetics 171: Shi YY and He L (2005). SHEsis, a powerful software platform for analyses of linkage disequilibrium, haplotype construction, and genetic association at polymorphism loci. Cell Res. 15:
9 X.P. An et al Smith JT, Cunningham MJ, Rissman EF, Clifton DK, et al. (2005). Regulation of Kiss1 gene expression in the brain of the female mouse. Endocrinology 146: Smith JT, Popa SM, Clifton DK, Hoffman GE, et al. (2006). Kiss1 neurons in the forebrain as central processors for generating the preovulatory luteinizing hormone surge. J. Neurosci. 26: Smith JT, Li Q, Yap KS, Shahab M, et al. (2011). Kisspeptin is essential for the full preovulatory LH surge and stimulates GnRH release from the isolated ovine median eminence. Endocrinology 152: Tomikawa J, Homma T, Tajima S, Shibata T, et al. (2010). Molecular characterization and estrogen regulation of hypothalamic KISS1 gene in the pig. Biol. Reprod. 82: Wang PQ, Deng LM, Zhang BY, Chu MX, et al. (2011). Polymorphisms of the cocaine-amphetamine-regulated transcript (CART) gene and their association with reproductive traits in Chinese goats. Genet. Mol. Res. 10: Xu N, Chen CY and Shyu AB (1997). Modulation of the fate of cytoplasmic mrna by AU-rich elements: key sequence features controlling mrna deadenylation and decay. Mol. Cell Biol. 17:
KISSPEPTIN AND GNIH CONTROL OF GNRH IN FEMALE MAMMALS M.J. Zamiri Department of Animal Science, College of Agriculture, Shiraz University, Shiraz, Iran firstname.lastname@example.org Introduction Since the discovery
Kisspeptin and other neuropeptides Higher centres Timing of puberty Stress New opportunities for reproductive endocrinology E2 +ve E2 -ve Richard A Anderson Obstetrics and Gynaecology University of Edinburgh
LH and FSH By Ronald Steriti, ND, PhD 2011 Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) are gonadotropins that stimulate the gonads - the testes in males, and the ovaries in females.
Investigation on ERCC5 genetic polymorphisms and the development of gastric cancer in a Chinese population L.Q. Yang 1, Y. Zhang 2 and H.F. Sun 3 1 Department of Gastroenterology, The Second Affiliated
FSH (Rodent) ELISA Kit Catalog Number KA2330 96 assays Version: 06 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 Principle of the Assay...
LH (Bovine) ELISA Kit Catalog Number KA2280 96 assays Version: 05 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 Principle of the Assay...
The menstrual Cycle François Pralong Services d Endocrinologie, Diabétologie et Métabolisme, Hôpitaux Universitaires de Genève et Lausanne Centre des Maladies CardioVasculaires et Métaboliques, Lausanne
Reproductive Health and Pituitary Disease Janet F. McLaren, MD Assistant Professor Division of Reproductive Endocrinology and Infertility Department of Obstetrics and Gynecology email@example.com Objectives
The refinement of the pulsatile Luteinizing Hormone (LH) profile following pubertal maturation: a potential mechanism for enhanced recruitment of ovarian follicles for improved fertility Ying Wan Master
1. The graph shows the changes in concentration of the hormones responsible for controlling the menstrual cycle. A Hormone concentration Oestrogen B C 0 14 28 Time / days WD Phillips and TJ Chilton A Level
Am J Physiol Endocrinol Metab 297: E1212 E1221, 2009. First published September 15, 2009; doi:10.1152/ajpendo.00461.2009. Sex differences in the regulation of Kiss1/NKB neurons in juvenile mice: implications
Identification of a novel duplication mutation in the VHL gene in a large Chinese family with Von Hippel-Lindau (VHL) syndrome L.H. Cao 1, B.H. Kuang 2, C. Chen 1, C. Hu 2, Z. Sun 1, H. Chen 2, S.S. Wang
Role of XRCC1 gene polymorphisms in non-small cell lung cancer cisplatin-based chemotherapy, and their effect on clinical and pathological characteristics H.F. Liu, J.S. Liu, J.H. Deng and R.R. Wu Department
Federal Employee Program 1310 G Street, N.W. Washington, D.C. 20005 202.942.1000 Fax 202.942.1125 5.30.03 Subject: Synarel Page: 1 of 5 Last Review Date: September 15, 2016 Synarel Description Assisted
Concentrations of Luteinizing Hormone and Ovulatory Responses in Dairy Cows Before Timed Artificial Insemination S. L. Pulley, D. H. Keisler, S. L. Hill, and J. S. Stevenson Summary The objective of this
Reproductive System (Hormone Function) Physiology Department Medical School, University of Sumatera Utara 1 Endocrine Control: Three Levels of Integration Hormones of the hypothalamic-anterior pituitary
Edinburgh Research Explorer Neurokinin 3 receptor antagonism decreases gonadotropin and testosterone secretion in healthy men Citation for published version: Skorupskaite, K, Jyothis, GT, Veldhuis, JD,
www.springerlink.co Chin J Cancer Res 22(1):49-54, 2010 49 Original Article Polymorphisms of XPC Gene And Susceptibility of Esophageal Cancer Xiang-xian Feng 1, 2, Pei-fen Duan 2, Li-bing Wang 2, Zu-xun
The Human Major Histocompatibility Complex 1 Location and Organization of the HLA Complex on Chromosome 6 NEJM 343(10):702-9 2 Inheritance of the HLA Complex Haplotype Inheritance (Family Study) 3 Structure
ACUPUNCTURE AND FEMALE INFERTILITY About infertility Infertility is the inability of a couple to get pregnant despite having regular unprotected sex. A couple is regarded as infertile if, after regular
ORIGINAL ARTICLE Pediatrics http://dx.doi.org/10.3346/jkms.2014.29.8.1120 J Korean Med Sci 2014; 29: 1120-1125 KISS1 Gene Polymorphisms in Korean Girls with Central Precocious Puberty Young-Jun Rhie, 1
REPRODUCTION-DEVELOPMENT Insulin-Like Growth Factor-I Activates KiSS-1 Gene Expression in the Brain of the Prepubertal Female Rat Jill K. Hiney, Vinod K. Srivastava, Michelle D. Pine, and W. Les Dees Department
International Journal of Reproduction, Contraception, Obstetrics and Gynecology Bonyadi K et al. Int J Reprod Contracept Obstet Gynecol. 2017 Sep;6(9):3760-3764 www.ijrcog.org DOI: http://dx.doi.org/10.18203/2320-1770.ijrcog20174021
J Med Dent Sci 2003; 50: 225 229 Original Article C18orf1 located on chromosome 18p11.2 may confer susceptibility to schizophrenia Mika Kikuchi 1,2, Kazuo Yamada 1, Tomoko Toyota 1,2 and Takeo Yoshikawa
FERTILITY AND STERILITY VOL. 80, NO. 4, OCTOBER 2003 Copyright 2003 American Society for Reproductive Medicine Published by Elsevier Inc. Printed on acid-free paper in U.S.A. CASE REPORTS Preimplantation
Chapter 28: REPRODUCTIVE SYSTEM: MALE I. FUNCTIONAL ANATOMY (Fig. 28.1) A. Testes: glands which produce male gametes, as well as glands producing testosterone 2. Seminiferous tubules (Fig.28.3; 28.5) a.
MCP-1 gene polymorphisms in North Chinese patients with pulmonary tuberculosis G.L. Shi, L. Yang, Y. Sun, Y.J. Yin and C.X. Song Department of Clinical Immunology Laboratory, Beijing Tuberculosis and Thoracic
Bio 111 Study Guide Chapter 17 From Gene to Protein BEFORE CLASS: Reading: Read the introduction on p. 333, skip the beginning of Concept 17.1 from p. 334 to the bottom of the first column on p. 336, and
Chapter 2 Gene and Promoter Structures of the Dopamine Receptors Ursula M. D Souza Abstract The dopamine receptors have been classified into two groups, the D 1 - like and D 2 -like dopamine receptors,
DOI:http://dx.doi.org/10.7314/APJCP.2013.14.9.5269 RESEARCH ARTICLE Tao Zhang*, Hui Wang Abstract Aim: We conducted a case-control matched study to investigate the role of IL-16 gene polymorphisms, rs4072111,
Investigation: The Human Menstrual Cycle Research Question: How do hormones control the menstrual cycle? Introduction: The menstrual cycle (changes within the uterus) is an approximately 28-day cycle that
Low levels of serum mir-99a is a predictor of poor prognosis in breast cancer J. Li 1, Z.J. Song 2, Y.Y. Wang 1, Y. Yin 1, Y. Liu 1 and X. Nan 1 1 Tumor Research Department, Shaanxi Provincial Tumor Hospital,
THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 272, No. 29, Issue of July 18, pp. 18060 18070, 1997 1997 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in U.S.A. Two Distinct TATA-less
ORIGINAL ARTICLE Pediatrics DOI: 10.3346/jkms.2011.26.7.927 J Korean Med Sci 2011; 26: 927-931 Serum Kisspeptin Levels in Korean Girls with Central Precocious Puberty Young Jun Rhie 1,2, Kee Hyoung Lee
Effect of variation of ABCB1 and GSTP1 on osteosarcoma survival after chemotherapy J.Z. Li 1, Z.Q. Tian 2, S.N. Jiang 3 and T. Feng 4 1 Department of Orthopedics, Juye County People s Hospital, Heze, China
Blocking the expression of the hepatitis B virus S gene in hepatocellular carcinoma cell lines with an anti-gene locked nucleic acid in vitro Y.-B. Deng, H.-J. Qin, Y.-H. Luo, Z.-R. Liang and J.-J. Zou
Action of reproductive hormones through the life span Do reproductive hormones affect the life span? One hypothesis about the rate of aging asserts that there is selective pressure for either high rate
Chinese Medicine, 2009, 1, 36-41 Published Online September 2009 in SciRes (www.scirp.org/journal/cm) Association of CALCA Genetic Polymorphism with Essential Hypertension ABSTRACT Calcitonin gene-related
UC San Diego UC San Diego Electronic Theses and Dissertations Title Ventral Anterior Homeodomain Protein Regulates Gonadotropin-Releasing Hormone and Fertility / Permalink https://escholarship.org/uc/item/89p7j1mq
Genetic variations in the Wnt signaling pathway affect lung function in asthma patients S.-H. Wang 1,2, F. Xu 1, H.-X. Dang 1 and L. Yang 1 1 Pediatric Intensive Care Unit, Children s Hospital, Chongqing
GONADAL FUNCTION: An Overview University of PNG School of Medicine & Health Sciences Division of Basic Medical Sciences Clinical Biochemistry BMLS III & BDS IV VJ Temple 1 What are the Steroid hormones?
814 Biomed Environ Sci, 2016; 29(11): 814-817 Letter to the Editor Association of TCF7L2 and GCG Gene Variants with Insulin Secretion, Insulin Resistance, and Obesity in New-onset Diabetes * ZHANG Lu 1,^,
The Open Breast Cancer Journal, 2011, 3, 1-5 1 Open Access Influence of RNA Interference Targeting Rab5a on Proliferation and Invasion of Breast Cancer Cell Line MCF-7 Nier Cha 1,2, Xinyue Gu 1, Fengyun
Veterinary Research Communications,28(2004) 111 119 2004 Kluwer Academic Publishers. Printed in the Netherlands Manipulation of Ovarian Function for the Reproductive Management of Dairy Cows W.W. Thatcher1*,
The Reproductive System Part 3 Gamete Formation Introduction Chromosomes carry genetic information In humans, cells contain 46 chromosomes Gametes carry only 23 chromosomes Meiosis Special type of cell
1 Roadmap Quantitative traits What kinds of variation can selection work on? How much will a population respond to selection? Heritability How can response be restored? Inbreeding How inbred is a population?
5th World Congress on Ovulation Induction Roma, 13 15 September 2007 FSH pharmacogenetics Hermann M. Behre Center of Reproductive Medicine and Andrology, University Hospital Halle, Germany & Manuela Simoni
EGFR gene polymorphisms -216G>T and -191C>A are risk markers for gastric cancer in Mexican population J.H. Torres-Jasso 1,2, M.E. Marín 3, E. Santiago-Luna 4, J.C. Leoner 5, J. Torres 6, M.T. Magaña-Torres
MOLECULAR AND CELLULAR BIOLOGY, June 1996, p. 3012 3022 Vol. 16, No. 6 0270-7306/96/$04.00 0 Copyright 1996, American Society for Microbiology Base Pairing at the 5 Splice Site with U1 Small Nuclear RNA
0013-7227/05/$15.00/0 Endocrinology 146(9):3917 3925 Printed in U.S.A. Copyright 2005 by The Endocrine Society doi: 10.1210/en.2005-0337 Changes in Hypothalamic KiSS-1 System and Restoration of Pubertal
Original Article Association between hsa-mir-34b/c rs4938723 T > C promoter polymorphism and cancer risk: a meta-analysis based on 6,036 cases and 6,204 controls Tao Tao 1,2,3 *, Shuqiu Chen 1,2,3 *, Bin
Plasma LH and FSH concentrations in prepubertal beef heifers before and in response to repeated injections of low doses of Gn-RH B. J. McLeod, W. Haresign, A. R. Peters and G. E. Lamming A.R.C. Research
Characterization of Luteinizing Hormone and Luteinizing Hormone Receptor and Their Indispensable Role in the Ovulatory Process of the Medaka Katsueki Ogiwara 1, Chika Fujimori 1, Sanath Rajapakse 2, Takayuki
CORRECTION NOTICE Nat. Genet. 42, 759 763 (2010); published online 22 August 2010; corrected online 27 August 2014 Genome-wide association study of esophageal squamous cell carcinoma in Chinese subjects
Course Outline BGEN3020 Introduction to Human Genetics Course Coordinator: Dr. David Merz Department of Biochemistry and Medical Genetics Faculty of Health Sciences Rm 324 Basic Medical Sciences Building
CHAPTER 41: Animal Hormones 1. List a few similarities and differences comparing: a. endocrine system b. nervous system 2. What is the difference between endocrine and exocrine glands? 3. What is the difference
mix P241-D2 MODY mix 1 Lot D2-0413. As compared to version D1 (lot D1-0911), one reference has been replaced. Maturity-Onset Diabetes of the Young (MODY) is a distinct form of non insulin-dependent diabetes
1 Effect of Resistin on Granulosa and Theca Cell Function in Cattle D.V. Lagaly, P.Y. Aad, L.B. Hulsey, J.A. Grado-Ahuir and L.J. Spicer Story in Brief Resistin is an adipokine that has not been extensively
AP 2 Exam Chapter 16 Endocrie Due Wed. night 4/22 or Thurs. morning 4/23 Name: Matching; match the labeled organ with the most appropriate response or identification. Figure 16.1 Using Figure 16.1, match
Female Reproduction 1 and 2: The Menstrual Cycle R.J. Witorsch, Ph.D. OBJECTIVES: At the end of this block of lectures, the student should be able to: 1. Describe the functional anatomy of ovary with particular
Overview of the Expressway Cell-Free Expression Systems The Expressway Cell-Free Expression Systems use an efficient coupled transcription and translation reaction to produce up to milligram quantities
Neurology Asia 2017; 22(3) : 221 225 Association of CALCA and RAMP1 gene polymorphisms with migraine in a Chinese population Xingkai An, Zhenzhen Yu, Jie Fang, Qing Lin, Congxia Lu, Qilin Ma, Hongli Qu
Chapter 18 Major endocrine glands and their hormones Endocrine glands Pituitary gland Has two major parts Anterior lobe called the adenohypophysis is epithelial in origin Posterior lobe called the neurohypophysis
ESM Methods Hyperinsulinemic-euglycemic clamp procedure During the hyperinsulinemic-euglycemic clamp , a priming dose of human insulin (Novolin, Clayton, NC) was followed by a constant rate (60 mu m
Viral Load DNA >> Standard PCR standard 0 Copies Catalog Number: 1122 Lot Number: 150298 Release Category: A Provided: 500 µl of 5.6 mm Tris HCl, 4.4 mm Tris base, 0.05% sodium azide 0.1 mm EDTA, 5 mg/liter
Global variation in copy number in the human genome Redon et. al. Nature 444:444-454 (2006) 12.03.2007 Tarmo Puurand Study 270 individuals (HapMap collection) Affymetrix 500K Whole Genome TilePath (WGTP)
European Journal of Endocrinology (2006) 155 S11 S16 ISSN 0804-4643 The role of KiSS-1 in the regulation of puberty in higher primates Tony M Plant Department of Cell Biology and Physiology and Obstetrics,
Am J Physiol Endocrinol Metab 296: E520 E531, 2009. First published January 13, 2009; doi:10.1152/ajpendo.90895.2008. KiSS-1 in the mammalian ovary: distribution of kisspeptin in human and marmoset and
The Endocrine System Like nervous system, endocrine system provides communication and control. Messages are relayed from one cell to another via chemical messengers (hormones). Unlike nervous system which