Iguchi Hirotoshi, Katsuzawa Takumi, Saruta Chihiro, Sadakata Tetsushi, Kobayashi Shota, Sato Yumi, Sato Akira, Sano Yoshitake, Maezawa So, Shinoda Yo, Furuichi Teiichi
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda, Japan.
Department of Environmental Health, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Japan.
Front Mol Neurosci. 2024 Jul 18;17:1444629. doi: 10.3389/fnmol.2024.1444629. eCollection 2024.
The Ca-dependent activator protein for secretion (CAPS/CADPS) family protein facilitates catecholamine release through the dense-core vesicle exocytosis in model neuroendocrine cell lines. However, it remains unclear if it induces dopamine release in the central neurons. This study aimed to examine the expression and function of CADPS2, one of the two CADPS paralogs, in dopamine neurons of the mouse midbrain. This study shows that CADPS2 was expressed in tyrosine hydroxylase and the vesicular monoamine transporter 2 (VMAT2)-positive dopaminergic neurons of the midbrain samples and primary mesencephalic cell cultures. Subcellular fractions rich in dopamine were collected using immunoaffinity for CADPS2 from midbrain protein extracts. Cell imaging using fluorescent false neurotransmitter FFN511 as a substrate for VMAT2 showed decreased activity-dependent dopamine release in -deficient cultures, compared to that in wild-type cultures. These results suggest that CADPS2 is involved in dopamine release from the central neurons, indicating its involvement in the central dopamine pathway.
分泌型钙依赖性激活蛋白(CAPS/CADPS)家族蛋白通过模型神经内分泌细胞系中的致密核心囊泡胞吐作用促进儿茶酚胺释放。然而,其是否能诱导中枢神经元释放多巴胺仍不清楚。本研究旨在检测CADPS两个旁系同源物之一CADPS2在小鼠中脑多巴胺能神经元中的表达及功能。本研究表明,CADPS2在中脑样本和原代中脑细胞培养物中酪氨酸羟化酶和囊泡单胺转运体2(VMAT2)阳性的多巴胺能神经元中表达。利用针对CADPS2的免疫亲和从脑蛋白提取物中收集富含多巴胺的亚细胞组分。使用荧光假神经递质FFN511作为VMAT2底物的细胞成像显示,与野生型培养物相比,CADPS2缺陷型培养物中活性依赖性多巴胺释放减少。这些结果表明,CADPS2参与中枢神经元的多巴胺释放,提示其参与中枢多巴胺能通路。