title: Molecular cloning and expression analysis of the synaptotagmin-1 gene in the hypothalamus and pituitary of Huoyan goose during different stages of the egg-laying cycle.
abstract: BACKGROUND
  Synaptotagmin-1 (Syt1) is an abundant, evolutionarily conserved integral membrane protein that plays essential roles in neurotransmitter release and hormone secretion. Neurotransmitters secreted by hypothalamic neurons can alter GnRH (gonadotropin-releasing hormones) neuronal activity by binding to and activating specific membrane receptors in pituitary cells and, in turn, control the release of gonadotropin hormones from the pituitary gland. To reveal the influence of Syt1 on the process of goose egg-laying, we cloned and characterized the cDNA of goose Syt1 originating from hypothalamus and pituitary tissues of Huoyan goose and investigated the mRNA expression profiles during different stages of the egg-laying cycle.
  
  
  METHODS
  Hypothalamus and pituitary tissues were obtained from 36 Huoyan geese in the pre-laying period, early laying period, peak-laying period, and ceased period. The cDNA sequences of goose Syt1 were cloned and characterized from Huoyan goose tissues using 5'-RACE and 3'-RACE methods. Multiple alignments and phylogenetic analyses of the deduced Syt1 amino acid sequence were conducted using bioinformatics tools. The expression profiles of the Syt1 mRNA in the hypothalamus and pituitary during pre-laying, early laying, peak-laying and ceased period were examined using real-time PCR (qRT-PCR).
  
  
  RESULTS
  The cDNA of Syt1 consisted of a 274 bp 5' UTR, a 1266 bp open reading frame (ORF) encoding 421 amino acids, and a 519 bp 3' UTR. The deduced amino acid sequence of goose Syt1 is highly conserved with the sequence from other species, especially with birds (more than 98%), and contains two protein kinase C2 conserved regions (C2 domain) from amino acids residue 157 to 259 and 288 to 402. The results of qRT-PCR demonstrated that the expression of Syt1 mRNA increased from the pre-laying period to the peak-laying period, reached its peak in the peak-laying period, and then decreased in the ceased period.
  
  
  CONCLUSIONS
  To the best of our knowledge, this study is the first to obtain full-length cDNA sequences of the goose Syt1 gene, and the results of Syt1 mRNA expression profiling in the hypothalamus and pituitary tissues suggested that Syt1 may play an important role in regulating the secretion of hormones relevant to the reproduction and egg-laying of female geese.
authors.affiliation: College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, China. xhluan@163.com.
authors.forename: Xinhong
authors.initials: X
authors.lastname: Luan
authors.forename: Lina
authors.initials: L
authors.lastname: Luo
authors.forename: Zhongzan
authors.initials: Z
authors.lastname: Cao
authors.forename: Rongrong
authors.initials: R
authors.lastname: Li
authors.forename: Dawei
authors.initials: D
authors.lastname: Liu
authors.forename: Ming
authors.initials: M
authors.lastname: Gao
authors.forename: Mei
authors.initials: M
authors.lastname: Liu
authors.forename: Laiyou
authors.initials: L
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030161:Avian Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods; D017124:Conserved Sequence; D005260:Female; D005777:Geese / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental; D007031:Hypothalamus / Q000378:metabolism*; D008969:Molecular Sequence Data; D010058:Oviposition; D010802:Phylogeny; D010902:Pituitary Gland / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D050863:Synaptotagmin I / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D030161:Avian Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D050863:Synaptotagmin I