Curry Haley Nicole, Huynh Roger, Rouhana Labib
Department of Biological Sciences, Wright State University, 3640 Colonel Glenn Hwy., Dayton, OH 45435, USA.
Department of Biology, University of Massachusetts Boston, 100 William T. Morrissey Blvd., Boston, MA 02125-3393, USA.
bioRxiv. 2024 Sep 3:2024.09.01.610670. doi: 10.1101/2024.09.01.610670.
The Transient Receptor Potential superfamily of proteins (TRPs) form cation channels that are abundant in animal sensory systems. Amongst TRPs, the Melastatin-related subfamily (TRPMs) is composed of members that respond to temperature, pH, sex hormones, and various other stimuli. Some TRPMs exhibit enriched expression in gonads of vertebrate and invertebrate species, but their contributions to germline development remain to be determined. We identified twenty-one potential TRPMs in the planarian flatworm and analyzed their anatomical distribution of expression by whole-mount hybridization. Enriched expression of two TRPMs ( and ) was detected in testis, whereas eight TRPM genes had detectable expression in patterns representative of neuronal and/or sensory cell types. Functional analysis of TRPM homologs by RNA-interference (RNAi) revealed that disruption of expression results in reduced sperm development, indicating a role for this receptor in supporting spermatogenesis. RNAi did not result in a detectable phenotype, but it increased sperm development deficiencies when combined with RNAi. Fluorescence hybridization revealed expression of in early spermatogenic cells within testes, suggesting cell-autonomous regulatory functions in germ cells for this gene. In addition, RNAi resulted in reduced numbers of presumptive germline stem cell clusters in asexual planarians, suggesting that supports establishment, maintenance, and/or expansion of spermatogonial germline stem cells. While further research is needed to identify the factors that trigger Smed-TRPM-c activity, these findings reveal one of few known examples for TRPM function in direct regulation of sperm development.
瞬时受体电位蛋白超家族（TRPs）形成阳离子通道，在动物感觉系统中大量存在。在TRPs中，与褪黑素相关的亚家族（TRPMs）由对温度、pH值、性激素和各种其他刺激有反应的成员组成。一些TRPMs在脊椎动物和无脊椎动物的性腺中表现出丰富的表达，但其对生殖系发育的贡献仍有待确定。我们在涡虫中鉴定出21种潜在的TRPMs，并通过整体杂交分析了它们的解剖学表达分布。在睾丸中检测到两种TRPMs（和）的丰富表达，而八个TRPM基因在代表神经元和/或感觉细胞类型的模式中有可检测到的表达。通过RNA干扰（RNAi）对TRPM同源物进行功能分析表明，的表达破坏会导致精子发育减少，表明该受体在支持精子发生中起作用。RNAi没有导致可检测到的表型，但与RNAi联合使用时会增加精子发育缺陷。荧光杂交显示在睾丸内早期生精细胞中有表达，表明该基因在生殖细胞中具有细胞自主调节功能。此外，RNAi导致无性涡虫中假定的生殖系干细胞簇数量减少，表明支持精原生殖系干细胞的建立、维持和/或扩增。虽然需要进一步研究来确定触发Smed-TRPM-c活性的因素，但这些发现揭示了TRPM在直接调节精子发育中的少数已知功能实例之一。
