机构:[1]State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, China.[2]European Molecular Biology Laboratory, European Bioinformatics Institute (EMBL-EBI), Wellcome Genome Campus, Cambridge CB10 1SD, United Kingdom.[3]Translational Research Institute of Brain and Brain-Like Intelligence and Department of Anesthesiology, Shanghai Fourth People's Hospital Affiliated to Tongji University School of Medicine, Shanghai, China.[4]Research Unit of Ocular Development and Regeneration, Chinese Academy of Medical Sciences, Beijing, China.[5]Guangdong Province Key Laboratory of Brain Function and Disease, Guangzhou, China.
Neurons are the bricks of the neuronal system and experimental access to certain neuron subtypes will be of great help to decipher neuronal circuits. Here, we identified trophoblast glycoprotein (TPBG)-expressing GABAergic amacrine cells (ACs) that were selectively labeled in DAT-tdTomato transgenic mice.Retina and brain sections were prepared for immunostaining with antibodies against various biomarkers. Patch-sequencing was performed to obtain the transcriptomes of tdTomato-positive cells in DAT-tdTomato mice. Whole-cell recordings were conducted to identify responses to light stimulation.Tyrosine hydroxylase immunoreactive cells were colocalized with tdTomato-positive cells in substantia nigra pars compacta, but not in the retina. Transcriptomes collected from tdTomato-positive cells in retinas via Patch-sequencing exhibited the expression of marker genes of ACs (Pax6 and Slc32a1) and marker genes of GABAergic neurons (Gad1, Gad2, and Slc6a1). Immunostaining with antibodies against relevant proteins (GAD67, GAD65, and GABA) also confirmed transcriptomic results. Furthermore, tdTomato-positive cells in retinas selectively expressed Tpbg, a marker gene for distinct clusters molecularly defined, which was proved with TPBG immunoreactivity in fluorescently labeled cells. Finally, tdTomato-positive cells recorded showed ON-OFF responses to light stimulation.Ectopic expression occurs in the retina but not in the substantia nigra pars compacta in the DAT-tdTomato mouse, and fluorescently labeled cells in the retina are TPBG-expressing GABAergic ACs. This type of transgenic mice has been proved as an ideal tool to achieve efficient labeling of a distinct subset of ACs that selectively express Tpbg.
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出版当年[2021]版:
大类|2 区医学
小类|2 区眼科学
最新[2025]版:
大类|2 区医学
小类|2 区眼科学
第一作者:
第一作者机构:[1]State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, China.
共同第一作者:
通讯作者:
通讯机构:[1]State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, China.[4]Research Unit of Ocular Development and Regeneration, Chinese Academy of Medical Sciences, Beijing, China.[5]Guangdong Province Key Laboratory of Brain Function and Disease, Guangzhou, China.[*1]Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China[*2]Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China[*3]Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China
推荐引用方式(GB/T 7714):
Huang Wanjing,Xu Qiang,Liu Feng,et al.Identification of TPBG-Expressing Amacrine Cells in DAT-tdTomato Mouse.[J].Investigative ophthalmology & visual science.2022,63(5):13.doi:10.1167/iovs.63.5.13.
APA:
Huang Wanjing,Xu Qiang,Liu Feng,Su Jing,Xiao Dongchang...&Liu Sheng.(2022).Identification of TPBG-Expressing Amacrine Cells in DAT-tdTomato Mouse..Investigative ophthalmology & visual science,63,(5)
MLA:
Huang Wanjing,et al."Identification of TPBG-Expressing Amacrine Cells in DAT-tdTomato Mouse.".Investigative ophthalmology & visual science 63..5(2022):13