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<p>Gonadotropin-releasing hormone plays a crucial role in vertebrate reproduction through the hypothalamo–hypophyseal–gonadal axis. While its functional effects are well understood, the transcriptional regulation of <i>gnrh1</i> remains poorly characterized in teleosts. This study investigates the promoter regulation of catfish <i>gnrh1,</i> i.e., <i>cfgnrh1</i>, in the brain of the Asian stinging catfish, <i>Heteropneustes fossilis</i>. To accomplish this, the 5′-upstream promoter fragments of <i>cfgnrh1</i> were isolated and characterized using in silico motif analysis, luciferase reporter assays, chromatin immunoprecipitation, and site-directed mutagenesis. The results identified Pax6 as a key regulator of <i>cfgnrh1</i> transcription. Immunolocalization of Pax6 and Gnrh in the preoptic area and hypothalamus of the <i>H. fossilis</i> brain indicates possible interaction. Polyethylenimine-mediated siRNA transient gene silencing of <i>pax6</i> confirmed its regulatory importance, as evidenced by the downregulation of <i>cfgnrh1</i>. Collectively, these findings highlight and validate a clear transcriptional interaction between <i>pax6</i> and <i>cfgnrh1</i>, emphasizing their significant roles in catfish reproduction.</p>

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Involvement of Pax6 in the transcriptional regulation of gnrh1 in the Asian stinging catfish Heteropneustes fossilis

  • 期刊论文

  • 发表日期：

    2026年5月23日

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  • Prabhaker Sangem,
  • Sonika Kar,
  • Swathi Tenugu,
  • Akanksha Pranoty,
  • Balasubramanian Senthilkumaran

摘要

Gonadotropin-releasing hormone plays a crucial role in vertebrate reproduction through the hypothalamo–hypophyseal–gonadal axis. While its functional effects are well understood, the transcriptional regulation of gnrh1 remains poorly characterized in teleosts. This study investigates the promoter regulation of catfish gnrh1, i.e., cfgnrh1, in the brain of the Asian stinging catfish, Heteropneustes fossilis. To accomplish this, the 5′-upstream promoter fragments of cfgnrh1 were isolated and characterized using in silico motif analysis, luciferase reporter assays, chromatin immunoprecipitation, and site-directed mutagenesis. The results identified Pax6 as a key regulator of cfgnrh1 transcription. Immunolocalization of Pax6 and Gnrh in the preoptic area and hypothalamus of the H. fossilis brain indicates possible interaction. Polyethylenimine-mediated siRNA transient gene silencing of pax6 confirmed its regulatory importance, as evidenced by the downregulation of cfgnrh1. Collectively, these findings highlight and validate a clear transcriptional interaction between pax6 and cfgnrh1, emphasizing their significant roles in catfish reproduction.