Guan Suhua, Huang Xin, Zhu Dongmei, Gao Zexia, Liu Han
Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education / Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture and Rural Affairs, College of Fisheries, Huazhong Agricultural University, Wuhan, China.
Hubei Hongshan Laboratory, Engineering Research Center of Green Development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt, Ministry of Education, Wuhan, China.
Front Cell Dev Biol. 2025 Sep 11;13:1587151. doi: 10.3389/fcell.2025.1587151. eCollection 2025.
Olfaction is essential for the survival and reproduction of fish, as it facilitates foraging, food localization, mate selection, and breeding. The cultured olfactory epithelial cells will provide an important resource for research on how fish use olfaction to detect odor molecules in their environment. In this study, olfactory epithelial cells from were cultured to investigate their responses to various odors, amino acids, and prostaglandin F (PGF). Initially, the olfactory epithelial cells were cultured using the explant method and collagenase digestion technique. Based on observations of growth characteristics, collagenase digestion demonstrated superior growth stability and morphological features of ciliated neurons. The presence of olfactory neurospheres was identified through scanning electron microscopy (SEM). Immunofluorescence analysis revealed that most of the cells cultured were labeled with NEUN antibody. Additionally, the expression of olfactory receptors () was detected in the cultured olfactory epithelial cells using fluorescence hybridization (FISH) and reverse transcription PCR (RT-PCR). Stimulation with amino acids mixture and PGF significantly increased the number of olfactory epithelial cells labeled with pERK. RNA-seq analysis revealed that 1,276 differentially expressed genes (DEGs) were identified following PGF stimulation, with pathways related to olfaction and reproduction being significantly enriched. Collectively, this study successfully established an model of the olfactory epithelium cells in and preliminarily investigated its response to odorant molecules, providing a valuable platform for research on fish olfactory function.
嗅觉对于鱼类的生存和繁殖至关重要，因为它有助于觅食、食物定位、配偶选择和繁殖。培养的嗅觉上皮细胞将为研究鱼类如何利用嗅觉检测环境中的气味分子提供重要资源。在本研究中，培养了[鱼类名称未给出]的嗅觉上皮细胞，以研究它们对各种气味、氨基酸和前列腺素F（PGF）的反应。最初，使用外植体方法和胶原酶消化技术培养嗅觉上皮细胞。基于对生长特征的观察，胶原酶消化显示出纤毛神经元具有更好的生长稳定性和形态特征。通过扫描电子显微镜（SEM）鉴定了嗅觉神经球的存在。免疫荧光分析表明，大多数培养的细胞用NEUN抗体标记。此外，使用荧光原位杂交（FISH）和逆转录PCR（RT-PCR）在培养的嗅觉上皮细胞中检测到嗅觉受体（）的表达。用氨基酸混合物和PGF刺激显著增加了用pERK标记的嗅觉上皮细胞数量。RNA测序分析显示，PGF刺激后鉴定出1276个差异表达基因（DEG），与嗅觉和繁殖相关的途径显著富集。总体而言，本研究成功建立了[鱼类名称未给出]嗅觉上皮细胞模型，并初步研究了其对气味分子的反应，为鱼类嗅觉功能研究提供了有价值的平台。
