Characterization of green fluorescent protein–expressing retinal cone bipolar cells in a 5-hydroxytryptamine receptor 2a transgenic mouse line
Pages 662-668
Q. Lu, E. Ivanova, Z.-H. Pan

Volume 163, Issue 2, Pages 662-668 (6 October 2009)

Abstract:Retinal bipolar cells relay visual information from photoreceptors to third-order retinal neurons. Bipolar cells, comprising multiple types, play an essential role in segregating visual information into multiple parallel pathways in the retina. The identification of molecular markers that can label specific retinal bipolar cells could facilitate the investigation of bipolar cell functions in the retina. Transgenic mice with specific cell type(s) labeled with green fluorescent protein (GFP) have become a powerful tool for morphological and functional studies of neurons in the CNS, including the retina. In this study, we report a 5-hydroxytryptamine receptor 2a (5-HTR2a) transgenic mouse line in which expression of GFP was observed in two populations of bipolar cells in the retina. Based on the terminal stratification and immunostaining, all the strongly GFP-labeled bipolar cells were found to be type 4 cone bipolar cells. A small population of weakly labeled bipolar cells was also observed, which may represent type 8 or 9 cone bipolar cells. GFP expression in retinal cone bipolar cells was seen as early as postnatal day 5. In addition, despite severe retinal degeneration due to the presence of the rd1 mutation in this transgenic line, the density of GFP-labeled cone bipolar cells remained stable up to at least 6 months of age. This transgenic mouse line will be a useful tool for the study of type 4 cone bipolar cells in the retina under both normal and disease conditions.

GENESAT(genesat.arc.nasa.gov)是一个Howard Hughes Medical Institute, 和The Rockefeller University合作的一个project,就是把GFP接在各种基因的promoter后面,做转基因小鼠.然后看看GFP在哪些细胞里表达.因为之前有一种转基因小鼠的GFP特异性表达在type5 bipolar cell里,而这个基因是5-HT3 receptor.刚好学校有位老师用5-HT2a-GFP的鼠研究大脑神经元.我的导师潘卓华教授就想看看在视网膜里如何.结果在type4 bipolar cell里有非常强而且单一的表达.试了很多种抗体.

同时Switzerland也有一个大老板Botond Roska在做,而且他是大批量的做,所以潘老师有点赶.如果在他们后面就没有意义了.他们做了几百种,我只做一种,当然没他们有意义了.他们的文章发表在Nature Neuroscience上:

Genetic address book for retinal cell types

Sandra Siegert1, Brigitte Gross Scherf1, Karina Del Punta2, Nick Didkovsky2, Nathaniel Heintz2,3 &
Botond Roska

不过还好他们online发表是在8月2号,我们实验室的工作online发表在7月份.

最想感谢的是手把手教我做实验的Dr. Elena Ivanova, 来自俄罗斯的超级pp超级cute的mm,德国马普Marc Plank的PhD.她做事认真细致,非常严谨.我做错事她都很严厉的立刻指出来.在我身上投入了很多心血和精力.6月份我回国,这篇文章还有好几张图的实验是她帮我完成的.嗯,导师潘教授就不说了,大恩不言谢........我只说他是一位非常非常理想的老师+老板,能在这个实验室读研已经非常心满意足了.

还有同级的美国女生Amy Hinkel,封面的图是她设计的.作图软件Photoshop她比我熟太多了.刚好她在潘老师实验室做rotation,而我在另一个实验室做rotation.

 

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