Department of Otorhinolaryngology-Head and Neck Surgery, Zhongda Hospital, Southeast University, Nanjing, China.
Anhui Province Key Laboratory of Biological Macro-Molecules Research, Wannan Medical College, Wuhu, China.
J Histochem Cytochem. 2022 Aug;70(8):583-596. doi: 10.1369/00221554221119543. Epub 2022 Aug 16.
Herein, we aimed to use double-labeling immunofluorescence to describe the expression pattern of Calbindin-D28K (CaBP28K) in the mouse cochlea from late embryonic (E) stages to the adulthood. CaBP28K was expressed in the inner hair cells (IHCs) and the greater epithelial ridge (GER) at E17. In addition, its expression was observed in the interdental cells. On postnatal day 1 (P1), CaBP28K immunoreactivity was observed in the IHCs and outer hair cells (OHCs) and was also specifically expressed in the nucleus and the cytoplasm of spiral ganglion neurons (SGNs). At P8, CaBP28K labeling disappeared from the interdental cells, and the CaBP28K-positive domain within the GER shifted from the entire cytoplasm to only the apical and basal regions. At P14, CaBP28K immunoreactivity was lost from the GER; however, its expression in the IHCs and OHCs, as well as the SGNs, persisted into adulthood. The identification of CaBP28K in the hair cells (HCs) and cuticular plates, as well as SGNs, was confirmed by its colocalization with several markers for Sox2, Myosin VIIa, Phalloidin, and Tuj1. We also detected colocalization with calmodulin in the cytoplasm of both HCs and SGNs. Western blot revealed an increase in CaBP28K postnatal expression in the mouse cochlea.
在此,我们旨在使用双重免疫荧光标记来描述钙结合蛋白 28k(CaBP28K)在小鼠耳蜗中的表达模式,范围从胚胎晚期(E)阶段到成年期。在 E17 时,CaBP28K 在内耳毛细胞(IHC)和上皮嵴(GER)中表达。此外,还观察到其在牙间细胞中的表达。在出生后第 1 天(P1),CaBP28K 免疫反应性见于 IHC 和外毛细胞(OHC),并且在螺旋神经节神经元(SGN)的核和细胞质中也特异性表达。在 P8 时,牙间细胞中的 CaBP28K 标记消失,而 GER 中的 CaBP28K 阳性区域从整个细胞质转移到仅顶端和基底区域。在 P14 时,GER 中的 CaBP28K 免疫反应性丧失;然而,其在 IHC 和 OHC 以及 SGN 中的表达持续到成年期。CaBP28K 在毛细胞(HCs)和角质板以及 SGN 中的鉴定通过其与 Sox2、Myosin VIIa、鬼笔环肽和 Tuj1 的几种标志物的共定位得到证实。我们还检测到细胞质中的钙调蛋白与 HCs 和 SGN 共定位。Western blot 显示小鼠耳蜗中 CaBP28K 的表达在出生后增加。