Molecular Cloning and Identification of the Transcriptional Regulatory Domain of the Goat Neurokinin B Gene <i>TAC3</i>

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Author(s)

    • SUETOMI Yuta SUETOMI Yuta
    • Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
    • MATSUDA Fuko MATSUDA Fuko
    • Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
    • MAEDA Kei-ichiro
    • Laboratory of Animal Breeding, Department of Veterinary Medical Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan
    • TSUKAMURA Hiroko
    • Laboratory of Reproductive Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
    • OHKURA Satoshi
    • Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan

Abstract

Neurokinin B (NKB), encoded by <i>TAC3</i>, is thought to be an important accelerator of pulsatile gonadotropin-releasing hormone release. This study aimed to clarify the transcriptional regulatory mechanism of goat <i>TAC3</i>. First, we determined the full-length mRNA sequence of goat <i>TAC3</i> from the hypothalamus to be 820 b, including a 381 b coding region, with the putative transcription start site located 143-b upstream of the start codon. The deduced amino acid sequence of NKB, which is produced from preproNKB, was completely conserved among goat, cattle, and human. Next, we cloned 5'-upstream region of goat <i>TAC3</i> up to 3400 b from the translation initiation site, and this region was highly homologous with cattle <i>TAC3</i> (89%). We used this goat <i>TAC3</i> 5'-upstream region to perform luciferase assays. We created a luciferase reporter vector containing DNA constructs from –2706, –1837, –834, –335, or –197 to +166 bp (the putative transcription start site was designated as +1) of goat <i>TAC3</i> and these were transiently transfected into mouse hypothalamus-derived N7 cells and human neuroblastoma-derived SK-N-AS cells. The luciferase activity gradually increased with the deletion of the 5'-upstream region, suggesting that the transcriptional suppressive region is located between –2706 and –336 bp and that the core promoter exists downstream of –197 bp. Estradiol treatment did not lead to significant suppression of luciferase activity of any constructs, suggesting the existence of other factor(s) that regulate goat <i>TAC3</i> transcription.

Journal

  • Journal of Reproduction and Development

    Journal of Reproduction and Development 59(5), 463-469, 2013

    THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT

Codes

  • NII Article ID (NAID)
    130003361048
  • NII NACSIS-CAT ID (NCID)
    AA10936678
  • Text Lang
    ENG
  • ISSN
    0916-8818
  • NDL Article ID
    024943079
  • NDL Call No.
    Z54-H305
  • Data Source
    NDL  J-STAGE 
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