Parisi Valentina, Guerrera Maria C, Abbate Francesco, Garcia-Suarez Olivia, Viña Eliseo, Vega Jose A, Germanà Antonino
Dipartimento di Scienze Veterinarie (Sezione di Morfologia), Università di Messina, Italy.
Departamento de Morfología y Biología Celular, Grupo SINPOS, Universidad de Oviedo, Spain.
Ann Anat. 2014 Jul;196(4):178-82. doi: 10.1016/j.aanat.2014.01.004. Epub 2014 Mar 12.
The fish sensory epithelium contains three types of sensory cells denominated ciliated, microvillous, and crypt neurons. Each one differs from the other in its morphological, ultrastructural and molecular features, as well as in their projections to the central nervous system. Crypt neurons are present in both bony and cartilaginous fish and can be identified on the basis of their morphology and the expression of some specific proteins and genes. In this study we have investigated the morphology of crypt neurons, as well as the occurrence and co-localization of S100 protein, calretinin and TRPV4, three proposed markers for crypt cells, in the olfactory epithelium of adult zebrafish (Danio rerio) using double immunofluorescence associated to laser confocal microscopy. A sparse population of superficial S100 protein positive cells was detected being identified as crypt neurons. The calretinin immunoreactive cells were more abundant, occasionally resembling the morphology of the crypt cells but never displaying co-localization of both proteins. The TRPV4 positive cells differed in morphology from crypt cells, thus excluding the occurrence of TRPV4 in those cells. These results demonstrate that only S100 protein immunoreactivity can be used to identify crypt cells. Because some calretinin positive cells showed localization and morphology similar to the crypt cells of the sensory epithelium, the occurrence of two subtypes of crypt cells, S100 protein and calretinin positive, cannot be excluded. The significance of these findings remains to be elucidated.
鱼类感觉上皮包含三种类型的感觉细胞，分别称为纤毛神经元、微绒毛神经元和隐窝神经元。每一种在形态、超微结构和分子特征以及它们向中枢神经系统的投射方面都彼此不同。隐窝神经元存在于硬骨鱼和软骨鱼中，可以根据其形态以及一些特定蛋白质和基因的表达来识别。在本研究中，我们使用与激光共聚焦显微镜相关的双重免疫荧光技术，研究了成年斑马鱼（Danio rerio）嗅觉上皮中隐窝神经元的形态，以及三种被提议作为隐窝细胞标志物的S100蛋白、钙视网膜蛋白和瞬时受体电位香草酸亚型4（TRPV4）的出现情况和共定位。检测到少量浅表S100蛋白阳性细胞，被鉴定为隐窝神经元。钙视网膜蛋白免疫反应性细胞更为丰富，偶尔类似于隐窝细胞的形态，但从未显示出两种蛋白的共定位。TRPV4阳性细胞在形态上与隐窝细胞不同，因此排除了TRPV4在这些细胞中的存在。这些结果表明，只有S100蛋白免疫反应性可用于识别隐窝细胞。由于一些钙视网膜蛋白阳性细胞显示出与感觉上皮隐窝细胞相似的定位和形态，因此不能排除存在S100蛋白阳性和钙视网膜蛋白阳性这两种隐窝细胞亚型。这些发现的意义仍有待阐明。
