Brann Jessica H, Dennis John C, Morrison Edward E, Fadool Debra A
Program in Neuroscience and Molecular Biophysics, Biomedical Research Facility, Department of Biological Science, Florida State University, Tallahassee, Florida 32306, USA.
J Neurochem. 2002 Dec;83(6):1452-60. doi: 10.1046/j.1471-4159.2002.01266.x.
The vomeronasal organ (VNO) is the receptor portion of the accessory olfactory system and transduces chemical cues that identify social hierarchy, reproductive status, conspecifics and prey. Signal transduction in VNO neurons is apparently accomplished via an inositol 1,4,5-trisphosphate (IP3)-activated calcium conductance that includes a different set of G proteins than those identified in vertebrate olfactory sensory neurons. We used immunohistochemical (IHC) and SDS-PAGE/western analysis to localize three IP3 receptors (IP3R) in the rat VNO epithelium. Type-I IP3R expression was weak or absent. Antisera for type-II and -III IP3R recognized appropriate molecular weight proteins by SDS-PAGE, and labeled protein could be abolished by pre-adsorption of the respective antibody with antigenic peptide. In tissue sections, type-II IP3R immunoreactivity was present in the supporting cell zone but not in the sensory cell zone. Type-III IP3R immunoreactivity was present throughout the sensory zone and overlapped that of transient receptor potential channel 2 (TRPC2) in the microvillar layer of sensory epithelium. Co-immunoprecipitation of type-III IP3R and TRPC2 from VNO lysates confirmed the overlapping immunoreactivity patterns. The protein-protein interaction complex between type-III IP3R and TRPC2 could initiate calcium signaling leading to electrical signal production in VNO neurons.
犁鼻器(VNO)是副嗅觉系统的感受器部分,可转导识别社会等级、生殖状态、同种个体和猎物的化学信号。VNO神经元中的信号转导显然是通过一种由肌醇1,4,5-三磷酸(IP3)激活的钙电导来完成的,该钙电导包含一组与脊椎动物嗅觉感觉神经元中所鉴定的不同的G蛋白。我们使用免疫组织化学(IHC)和SDS-PAGE/蛋白质免疫印迹分析来定位大鼠VNO上皮中的三种IP3受体(IP3R)。I型IP3R表达较弱或缺失。II型和III型IP3R的抗血清通过SDS-PAGE识别出合适分子量的蛋白质,并且标记的蛋白质可以通过用抗原肽预先吸附相应抗体来消除。在组织切片中,II型IP3R免疫反应性存在于支持细胞区,而不存在于感觉细胞区。III型IP3R免疫反应性存在于整个感觉区,并与感觉上皮微绒毛层中的瞬时受体电位通道2(TRPC2)的免疫反应性重叠。从VNO裂解物中共免疫沉淀III型IP3R和TRPC2证实了重叠的免疫反应模式。III型IP3R和TRPC2之间的蛋白质-蛋白质相互作用复合物可以启动钙信号传导,导致VNO神经元产生电信号。