Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, United States of America.
PLoS One. 2020 Dec 11;15(12):e0243756. doi: 10.1371/journal.pone.0243756. eCollection 2020.
Reproduction is highly sensitive to changes in physiology and the external environment. Neuropeptides are evolutionarily conserved signaling molecules that regulate multiple physiological processes. However, the potential reproductive roles of many neuropeptide signaling pathways remain underexplored. Here, we describe the results of RNAi-based screens in Drosophila melanogaster to identify neuropeptides/neuropeptide receptors with potential roles in oogenesis. The screen read-outs were either the number of eggs laid per female per day over time or fluorescence microscopy analysis of dissected ovaries. We found that the orphan neuropeptide receptor encoded by moody (homologous to mammalian melatonin receptors) is likely required in somatic cells for normal egg production and proper germline stem cell maintenance. However, the egg laying screens had low signal-to-noise ratio and did not lead to the identification of additional candidates. Thus, although egg count assays might be useful for large-scale screens to identify oogenesis regulators that result in dramatic changes in oogenesis, more labor-intensive microscopy-based screen are better applicable for identifying new physiological regulators of oogenesis with more subtle phenotypes.
生殖对生理和外部环境的变化极为敏感。神经肽是进化上保守的信号分子，可调节多种生理过程。然而，许多神经肽信号通路的潜在生殖作用仍未得到充分探索。在这里，我们描述了在黑腹果蝇中进行的基于 RNAi 的筛选结果，以鉴定在卵子发生中具有潜在作用的神经肽/神经肽受体。筛选的读出值是每天每个雌性产卵的数量，或者是对解剖卵巢的荧光显微镜分析。我们发现，由 moody 编码的孤儿神经肽受体（与哺乳动物褪黑素受体同源）可能在体细胞中对于正常的卵子发生和正常的生殖干细胞维持是必需的。然而，产卵筛选的信噪比低，并未鉴定出其他候选物。因此，尽管卵计数测定可能适用于大规模筛选以鉴定导致卵子发生明显变化的卵子发生调节剂，但更耗时的基于显微镜的筛选更适用于鉴定具有更细微表型的卵子发生的新生理调节剂。
