Identification of TPBG-Expressing Amacrine Cells in DAT-tdTomato Mouse

Overview

Journal Invest Ophthalmol Vis Sci

Specialty Ophthalmology

Date 2022 May 13

PMID 35551574

Authors

Wanjing Huang

Qiang Xu

Feng Liu

Jing Su

Dongchang Xiao

Lei Tang

Zhao-Zhe Hao

Ruifeng Liu

Kangjian Xiang

Yalan Bi

Zhichao Miao

Xialin Liu

Yizhi Liu

Sheng Liu

Affiliations

Soon will be listed here.

Abstract

Purpose: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

Citing Articles

Comparative single-cell transcriptomic analysis reveals putative differentiation drivers and potential origin of vertebrate retina.

Zeng X, Gyoja F, Cui Y, Loza M, Kusakabe T, Nakai K NAR Genom Bioinform. 2024; 6(4):lqae149.

PMID: 39534499 PMC: 11555436. DOI: 10.1093/nargab/lqae149.

The RBPMS mouse line for studying retinal and vascular relevant diseases.

Li G, Luo Y, Zhang Q, Chen W, Lai K, Liu Y iScience. 2023; 26(11):108111.

PMID: 37867934 PMC: 10589894. DOI: 10.1016/j.isci.2023.108111.

References

Tran N, Shekhar K, Whitney I, Jacobi A, Benhar I, Hong G . Single-Cell Profiles of Retinal Ganglion Cells Differing in Resilience to Injury Reveal Neuroprotective Genes. Neuron. 2019; 104(6):1039-1055.e12. PMC: 6923571. DOI: 10.1016/j.neuron.2019.11.006. View

Peng Y, Shekhar K, Yan W, Herrmann D, Sappington A, Bryman G . Molecular Classification and Comparative Taxonomics of Foveal and Peripheral Cells in Primate Retina. Cell. 2019; 176(5):1222-1237.e22. PMC: 6424338. DOI: 10.1016/j.cell.2019.01.004. View

Liang C, Zhang G, Zhang L, Chen S, Wang J, Zhang T . Calmodulin Bidirectionally Regulates Evoked and Spontaneous Neurotransmitter Release at Retinal Ribbon Synapses. eNeuro. 2020; 8(1). PMC: 7808332. DOI: 10.1523/ENEURO.0257-20.2020. View

Stabio M, Sabbah S, Quattrochi L, Ilardi M, Fogerson P, Leyrer M . The M5 Cell: A Color-Opponent Intrinsically Photosensitive Retinal Ganglion Cell. Neuron. 2018; 97(1):251. PMC: 5769920. DOI: 10.1016/j.neuron.2017.12.030. View

Ullmann J, Moore B, Temple S, Fernandez-Juricic E, Collin S . The retinal wholemount technique: a window to understanding the brain and behaviour. Brain Behav Evol. 2011; 79(1):26-44. DOI: 10.1159/000332802. View

Zhang D, Zhou T, McMahon D . Functional heterogeneity of retinal dopaminergic neurons underlying their multiple roles in vision. J Neurosci. 2007; 27(3):692-9. PMC: 6672798. DOI: 10.1523/JNEUROSCI.4478-06.2007. View

Ran Y, Huang Z, Baden T, Schubert T, Baayen H, Berens P . Type-specific dendritic integration in mouse retinal ganglion cells. Nat Commun. 2020; 11(1):2101. PMC: 7193577. DOI: 10.1038/s41467-020-15867-9. View

Feng G, Mellor R, Bernstein M, Nguyen Q, Wallace M, Nerbonne J . Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron. 2000; 28(1):41-51. DOI: 10.1016/s0896-6273(00)00084-2. View

Kim I, Zhang Y, Meister M, Sanes J . Laminar restriction of retinal ganglion cell dendrites and axons: subtype-specific developmental patterns revealed with transgenic markers. J Neurosci. 2010; 30(4):1452-62. PMC: 2822471. DOI: 10.1523/JNEUROSCI.4779-09.2010. View

10.

Versaux-Botteri C, Nguyen-Legros J, Geffard M, Vigny A, Denoroy L . Regional specialization of the rat retina: catecholamine-containing amacrine cell characterization and distribution. J Comp Neurol. 1986; 243(3):422-33. DOI: 10.1002/cne.902430311. View

11.

Wakeham C, Ren G, Morgans C . Expression and distribution of trophoblast glycoprotein in the mouse retina. J Comp Neurol. 2020; 528(10):1660-1671. PMC: 7255440. DOI: 10.1002/cne.24850. View

12.

Zhang Y, Kim I, Sanes J, Meister M . The most numerous ganglion cell type of the mouse retina is a selective feature detector. Proc Natl Acad Sci U S A. 2012; 109(36):E2391-8. PMC: 3437843. DOI: 10.1073/pnas.1211547109. View

13.

Cadwell C, Scala F, Li S, Livrizzi G, Shen S, Sandberg R . Multimodal profiling of single-cell morphology, electrophysiology, and gene expression using Patch-seq. Nat Protoc. 2017; 12(12):2531-2553. PMC: 6422019. DOI: 10.1038/nprot.2017.120. View

14.

Martersteck E, Hirokawa K, Evarts M, Bernard A, Duan X, Li Y . Diverse Central Projection Patterns of Retinal Ganglion Cells. Cell Rep. 2017; 18(8):2058-2072. PMC: 5357325. DOI: 10.1016/j.celrep.2017.01.075. View

15.

Bruggen B, Meyer A, Boven F, Weiler R, Dedek K . Type 2 wide-field amacrine cells in TH::GFP mice show a homogenous synapse distribution and contact small ganglion cells. Eur J Neurosci. 2014; 41(6):734-47. DOI: 10.1111/ejn.12813. View

16.

Wassle H . Parallel processing in the mammalian retina. Nat Rev Neurosci. 2004; 5(10):747-57. DOI: 10.1038/nrn1497. View

17.

Macosko E, Basu A, Satija R, Nemesh J, Shekhar K, Goldman M . Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets. Cell. 2015; 161(5):1202-1214. PMC: 4481139. DOI: 10.1016/j.cell.2015.05.002. View

18.

Zhuang X, Masson J, Gingrich J, Rayport S, Hen R . Targeted gene expression in dopamine and serotonin neurons of the mouse brain. J Neurosci Methods. 2005; 143(1):27-32. DOI: 10.1016/j.jneumeth.2004.09.020. View

19.

Zeisel A, Munoz-Manchado A, Codeluppi S, Lonnerberg P, La Manno G, Jureus A . Brain structure. Cell types in the mouse cortex and hippocampus revealed by single-cell RNA-seq. Science. 2015; 347(6226):1138-42. DOI: 10.1126/science.aaa1934. View

20.

Madisen L, Zwingman T, Sunkin S, Oh S, Zariwala H, Gu H . A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci. 2009; 13(1):133-40. PMC: 2840225. DOI: 10.1038/nn.2467. View