Huang Liquan, Max Marianna, Margolskee Robert F, Su Hui, Masland Richard H, Euler Thomas
Department of Physiology and Biophysics, Mount Sinai School of Medicine, New York, New York 10029, USA.
J Comp Neurol. 2003 Jan 1;455(1):1-10. doi: 10.1002/cne.10396.
We investigated the expression of Ggamma13, a recently discovered G protein subunit, and a selection of Gbeta subunits in retinal bipolar cells, by using a transgenic mouse strain in which green fluorescent protein is strongly expressed in a single type of cone bipolar cell. The cells have ON morphology, and patch-clamp recordings in slices confirmed that they are of the physiological ON type. Immunohistochemistry showed that Ggamma13 is expressed in rod bipolar cells and ON cone bipolar cells, where it is colocalized in the dendrites with Galphaomicron. ON and OFF cone bipolar cells and rod bipolar cells were identified among dissociated cells by their green fluorescence and/or distinct morphology. Hybridization of single-cell polymerase chain reaction products with cDNA probes for G protein subunits Gbeta1 to 5 showed that Gbeta3, Gbeta4, and Ggamma13 are coexpressed in ON bipolar cells but not present in OFF bipolar cells. Gbeta1, 2, and 5 are expressed in partially overlapping subpopulations of cone bipolar cells. Ggamma13 and Gbeta3 and/or Gbeta4, thus, seem selectively to participate in signal transduction by ON bipolar cells.
我们利用一种转基因小鼠品系,研究了最近发现的G蛋白亚基Ggamma13以及一系列Gbeta亚基在视网膜双极细胞中的表达情况。在该转基因小鼠品系中,绿色荧光蛋白在单一类型的视锥双极细胞中强烈表达。这些细胞具有ON型形态,切片中的膜片钳记录证实它们属于生理上的ON型。免疫组织化学显示,Ggamma13在视杆双极细胞和ON视锥双极细胞中表达,在树突中与Galphaomicron共定位。通过绿色荧光和/或独特形态在解离细胞中鉴定出ON和OFF视锥双极细胞以及视杆双极细胞。单细胞聚合酶链反应产物与G蛋白亚基Gbeta1至5的cDNA探针杂交显示,Gbeta3、Gbeta4和Ggamma13在ON双极细胞中共表达,但在OFF双极细胞中不存在。Gbeta1、2和5在视锥双极细胞的部分重叠亚群中表达。因此,Ggamma13和Gbeta3和/或Gbeta4似乎选择性地参与ON双极细胞的信号转导。