Kerkhoven R M, Croll R P, Ramkema M D, Van Minnen J, Bogerd J, Boer H H
Department of Organismic Zoology, Faculty of Biology, Amsterdam, The Netherlands.
Cell Tissue Res. 1992 Mar;267(3):551-9. doi: 10.1007/BF00319378.
VD1 and RPD2 are two giant neuropeptidergic neurons in the central nervous system (CNS) of the pond snail Lymnaea stagnalis. We wished to determine whether other central neurons in the CNS of L. stagnalis express the VD1/RPD2 gene. To this end, in situ hybridization with the cDNA probe of the VD1/RPD2 gene and immunocytochemistry with antisera specific to VD1 and RPD2 (the alpha 1-antiserum, Mab4H5 and ALMA 6) and to R15 (the alpha 1 and 16-mer antisera) were performed on alternate tissue sections. A VD1/RPD2 neuronal system comprising three classes of neurons (A1-A3) was found. All neurons of the system express the gene. Division into classes is based on immunocytochemical characteristics. Class A1 neurons (VD1 and RPD2) immunoreact with the alpha 1-antiserum, Mab4H5 and ALMA 6. Class A2 neurons (1-5 small and 1-5 medium sized neurons in the visceral and right parietal ganglion, and two clusters of small neurons and 5 medium-sized neurons in the cerebral ganglia) immunoreact with the alpha 1-antiserum and Mab4H5, but not with ALMA 6. Class A3 neurons (3-4 medium-sized neurons and a cluster of 4-5 small neurons located in the pedal ganglion) immunoreact with the alpha 1-antiserum only. All neurons of the system are immunonegative to the R15 antisera. The observations suggest that the neurons of the VD1/RPD2 system produce different sets of neuropeptides. A group of approximately 15 neurons (class B), scattered in the ganglia, immunostained with one or more of the antisera, but did not react with the cDNA probe in in situ hybridization.
VD1和RPD2是椎实螺（Lymnaea stagnalis）中枢神经系统（CNS）中的两个巨大神经肽能神经元。我们希望确定椎实螺中枢神经系统中的其他中枢神经元是否表达VD1/RPD2基因。为此，我们在交替的组织切片上进行了VD1/RPD2基因cDNA探针的原位杂交，以及针对VD1和RPD2（α1抗血清、单克隆抗体4H5和抗Lymnaea促性腺激素抗体6）和R15（α1和16肽抗血清）的抗血清的免疫细胞化学实验。发现了一个由三类神经元（A1 - A3）组成的VD1/RPD2神经元系统。该系统的所有神经元都表达该基因。分类是基于免疫细胞化学特征。A1类神经元（VD1和RPD2）与α1抗血清、单克隆抗体4H5和抗Lymnaea促性腺激素抗体6发生免疫反应。A2类神经元（在内脏神经节和右顶神经节中有1 - 5个小神经元和1 - 5个中型神经元，在脑神经节中有两簇小神经元和5个中型神经元）与α1抗血清和单克隆抗体4H5发生免疫反应，但不与抗Lymnaea促性腺激素抗体6发生反应。A3类神经元（位于足神经节中的3 - 4个中型神经元和一簇4 - 5个小神经元）仅与α1抗血清发生免疫反应。该系统的所有神经元对R15抗血清均呈免疫阴性。这些观察结果表明，VD1/RPD2系统的神经元产生不同组的神经肽。一组约15个神经元（B类）散布在神经节中，用一种或多种抗血清进行免疫染色，但在原位杂交中不与cDNA探针反应。
