Jungblut Lucas D, Paz Dante A, López-Costa Juan J, Pozzi Andrea G
Laboratorio de Biología del Desarrollo, Instituto de Fisiología, Bioloíg Molecular y Neurociencias (IFIBYNE-CONICET) and Departamento de Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires.
Zoolog Sci. 2009 Oct;26(10):722-8. doi: 10.2108/zsj.26.722.
We evaluated the presence of G protein subtypes Galpha(o), Galpha(i2), and Galpha(olf) in the main olfactory system (MOS) and accessory or vomeronasal system (VNS) of Rhinella (Bufo) arenarum tadpoles, and here describe the fine structure of the sensory cells in the olfactory epithelium (OE) and vomeronasal organ (VNO). The OE shows olfactory receptor neurons (ORNs) with cilia in the apical surface, and the vomeronasal receptor neurons (VRNs) of the VNO are covered with microvilli. Immunohistochemistry detected the presence of at least two segregated populations of ORNs throughout the OE, coupled to Galpha(olf) and Galpha(o). An antiserum against Galpha(i2) was ineffective in staining the ORNs. In the VNO, Galpha(o) neurons stained strongly but lacked immunoreactivity to any other Galpha subunit in all larval stages analyzed. Western blot analyses and preabsorption experiments confirmed the specificity of the commercial antisera used. The functional significance of the heterogeneous G-protein distribution in R. arenarum tadpoles is not clear, but the study of G- protein distributions in various amphibian species is important, since this vertebrate group played a key role in the evolution of tetrapods. A more complete knowledge of the amphibian MOS and VNS would help to understand the functional organization and evolution of vertebrate chemosensory systems. This work demonstrates, for the first time, the existence of a segregated distribution of G-proteins in the OE of R. arenarum tadpoles.
我们评估了沙蟾蝌蚪主嗅觉系统(MOS)和副嗅觉或犁鼻器系统(VNS)中G蛋白亚型Gα(o)、Gα(i2)和Gα(olf)的存在情况,并在此描述嗅觉上皮(OE)和犁鼻器(VNO)中感觉细胞的精细结构。OE显示顶端表面有带纤毛的嗅觉受体神经元(ORN),VNO的犁鼻器受体神经元(VRN)则覆盖有微绒毛。免疫组织化学检测到整个OE中至少存在两个分离的ORN群体,分别与Gα(olf)和Gα(o)偶联。抗Gα(i2)抗血清对ORN染色无效。在VNO中,Gα(o)神经元染色强烈,但在所有分析的幼虫阶段对任何其他Gα亚基均无免疫反应性。蛋白质免疫印迹分析和预吸收实验证实了所用商业抗血清的特异性。沙蟾蝌蚪中异质G蛋白分布的功能意义尚不清楚,但研究各种两栖动物物种中的G蛋白分布很重要,因为这一脊椎动物群体在四足动物的进化中发挥了关键作用。对两栖动物MOS和VNS更全面的了解将有助于理解脊椎动物化学感觉系统的功能组织和进化。这项工作首次证明了沙蟾蝌蚪OE中存在G蛋白的分离分布。