Murphy F A, Tucker K, Fadool D A
Auburn University, Zoology and Wildlife Sciences, Auburn, Alabama 36849-5414, USA.
J Comp Neurol. 2001 Mar 26;432(1):61-74. doi: 10.1002/cne.1088.
We have explored the use of a new model to study the transduction of chemosignals in the vomeronasal organ (VNO), for which the functional pathway for chemical communication is incompletely understood. Because putative vomeronasal receptors in mammalian and other vertebrate models belong to the superfamily of G-protein-coupled receptors, the objective of the present study was to define which G-protein subunits were present in the VNO of Sternotherus odoratus (stinkpot or musk turtle) in order to provide directionality for future functional studies of the downstream signaling cascades. The turtle vomeronasal epithelium (VNE) was found to contain the G-proteins G(beta) and G(alphail-3) at the microvillar layer, the presumed site of signal tranduction in these neurons, as evidenced by immunocytochemical techniques. G(alphao) labeled the axon bundles in the VNE and the somata of the vomeronasal sensory neurons but not the microvillar layer. Densitometric analysis of Western blots indicated that the VNO from females contained greater concentrations of G(alphai1-3) compared with males. Sexually immature (juvenile) turtles showed intense immunolabeling for all three subunits (G(beta), G(alphai1-3), and G(alphao)) in the axon bundles and an absence of labeling in the microvillar layer. Another putative signaling component found in the microvilli of mammalian VNO, transient receptor potential channel, was also immunoreactive in S. odoratus in a gender-specific manner, as quantified by Western blot analysis. These data demonstrate the utility of Sternotherus for discerning the functional signal transduction machinery in the VNO and may suggest that gender and developmental differences in effector proteins or cellular signaling components may be used to activate sex-specific behaviors.
我们探索了使用一种新模型来研究犁鼻器(VNO)中化学信号的转导,其化学通讯的功能途径尚未完全了解。由于哺乳动物和其他脊椎动物模型中的假定犁鼻器受体属于G蛋白偶联受体超家族,本研究的目的是确定香龟(麝香龟)的犁鼻器中存在哪些G蛋白亚基,以便为下游信号级联反应的未来功能研究提供方向性。通过免疫细胞化学技术证明,在这些神经元中假定的信号转导位点——微绒毛层,发现龟的犁鼻器上皮(VNE)含有G蛋白G(β)和G(αi1 - 3)。G(αo)标记了VNE中的轴突束和犁鼻器感觉神经元的胞体,但未标记微绒毛层。蛋白质免疫印迹的光密度分析表明,与雄性相比,雌性的犁鼻器中G(αi1 - 3)的浓度更高。性未成熟(幼年)龟的轴突束中所有三个亚基(G(β)、G(αi1 - 3)和G(αo))均显示强烈的免疫标记,而微绒毛层未标记。通过蛋白质免疫印迹分析定量,在哺乳动物犁鼻器微绒毛中发现的另一种假定信号成分——瞬时受体电位通道,在香龟中也具有性别特异性的免疫反应性。这些数据证明了香龟在识别犁鼻器中功能信号转导机制方面的实用性,并可能表明效应蛋白或细胞信号成分的性别和发育差异可用于激活性别特异性行为。