Molecular codes for cell type specification in Brn3 retinal ganglion cells

Sajgo, Szilard; Ghinia, Miruna Georgiana; Brooks, Matthew; Kretschmer, Friedrich; Chuang, Katherine; Hiriyanna, Suja; Wu, Zhijian; Popescu, Octavian; Badea, Tudor C.

doi:10.1073/pnas.1618551114

Cited by 64 publications

(126 citation statements)

References 89 publications

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“…Intriguingly, we also detected the expression of Tbr1 , a T-brain subfamily gene closely related to Tbr2 , in mouse retinas (Mao et al, 2008, Naiche et al, 2005). A recent paper confirmed the expression of Tbr1 in RGCs (Sajgo et al, 2017). To trace the spatiotemporal expression of Tbr1 during retinogenesis and to study its functions during retinal development, we generated 2 knock-in mouse lines, Tbr1 TauGFP-IRESCreERT2 and Tbr1 mycflox alleles.…”

Section: Introductionmentioning

confidence: 61%

Essential Roles of Tbr1 in the Formation and Maintenance of the Orientation-Selective J-RGCs and a Group of OFF-Sustained RGCs in Mouse

et al. 2019

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SUMMARY In mouse retina, more than 30 retinal ganglion cell (RGC) subtypes have been classified based on a combined metric of morphological and functional characteristics. RGCs arise from a common pool of retinal progenitor cells during embryonic stages and differentiate into mature subtypes in adult retinas. However, the cellular and molecular mechanisms controlling the formation and maturation of such remarkable cellular diversity remain unknown. Here, we demonstrate that T-box transcription factor T-brain 1 (Tbr1) is expressed in 2 groups of morphologically and functionally distinct RGCs: the orientation-selective J-RGCs and a group of OFF-sustained RGCs with symmetrical dendritic arbors. When Tbr1 is genetically ablated during retinal development, these 2 RGC groups cannot develop. Ectopically expressing Tbr1 in M4 ipRGCs during development alters dendritic branching and density but not inner plexiform layer stratification level. Our data indicate that Tbr1 plays critical roles in regulating the formation and dendritic morphogenesis of specific RGC types.

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Section: Introductionmentioning

confidence: 61%

Essential Roles of Tbr1 in the Formation and Maintenance of the Orientation-Selective J-RGCs and a Group of OFF-Sustained RGCs in Mouse

et al. 2019

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“…To study and compare the expression profile of Copines both embryonically and postnatally, we extracted transcript and gene level expression data for the nine members of the Copine family from our previously reported RNA sequencing data for RGCs, retina and the retino recipient brain areas (Sajgo et al, ; Figure and Supporting Information Figure S1). Briefly, in our previous work, we had purified by immunomagnetic isolation genetically labeled RGCs expressing either Brn3a or Brn3b (Brn3a AP and Brn3b AP RGCs), and compared their RNA expression profile with expression profiles generated from age‐matched (postnatal day 3, P3) whole retina or retinorecipient brain areas.…”

Section: Resultsmentioning

confidence: 99%

“…Several Copine family members appeared as differentially regulated and/or expressed targets, and preliminary results in tissue culture showed a potential role for Cpne4 in cellular morphology and process extension. For details of the methods used for RNA sequencing, please refer to Sajgo et al ().…”

Section: Resultsmentioning

confidence: 99%

“…However, the molecules that link the combinatorial Brn3 expression to the activity‐dependent and/or a retrograde signaling pathway‐dependent pruning of dendrites are not known. Brn3a and 3b regulate the expression of several proteins in the mouse retina (Sajgo et al, ). Whether these molecules have a role in dendritic stratification, synapse formation, and axon targeting to the brain by themselves or in conjunction with combinatorial Brn3 expression remains to be explored.…”

Section: Introductionmentioning

confidence: 99%

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Differential expression and subcellular localization of Copines in mouse retina

Goel

Badea

2019

J of Comparative Neurology

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Combinatorial expression of Brn3 transcription factors is required for the development of cell‐specific morphologies in retinal ganglion cells (RGCs). The molecular mechanisms by which Brn3s regulate RGC type specific features are largely unexplored. We previously identified several members of the Copine (Cpne) family of molecules as potential targets of Brn3 transcription factors in the retina. We now use in situ hybridization and immunohistochemistry to characterize Copine expression in the postnatal and adult mouse retina. We find that Cpne5, 6, and 9 are expressed in the ganglion cell layer (GCL) and inner nuclear layer (INL) in both amacrine cells and RGCs. Cpne4 expression is restricted to one amacrine cell population of the INL, but is specifically expressed in RGCs in the GCL. Cpne4 expression in RGCs is regulated by Brn3b both cell autonomously (in Brn3b+ RGCs) and cell nonautonomously (in Brn3b− RGCs). Copines exhibit a variety of subcellular distributions when overexpressed in tissue culture cells (HEK293), and can induce the formation of elongated processes reminiscent of neurites in these non‐neuronal cells. Our results suggest that Copines might be involved in a combinatorial fashion in Brn3b‐dependent specification of RGC types. Given their expression profile and previously proven role as Ca2+ sensors, they may participate in the morphogenetic processes that shape RGC dendrite and axon formation at early postnatal ages.

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“…1 Because the dysfunction and impairment of RGCs in syndromes such as glaucoma and optic neuropathy lead to vision loss, the pathology and etiology of RGC-linked diseases have been investigated widely. For example, glaucoma is the second most common cause of blindness worldwide 2,3 and is characterized by RGC death caused by axonal degeneration.…”

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confidence: 99%

Culture Systems of Dissociated Mouse and Human Pluripotent Stem Cell–Derived Retinal Ganglion Cells Purified by Two-Step Immunopanning

Kobayashi

Onishi

et al. 2018

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. We aimed to establish purification and culture systems for retinal ganglion cells (RGCs) differentiated from mouse and human pluripotent stem cells (PSC) for in vitro and regenerative medicine studies. METHODS.We used a two-step immunopanning method to purify RGCs from mouse and human PSC-derived three-dimensional (3D) retinal organoids. To assess the method, we purified RGCs from 3D retinal organoids derived from embryonic stem cells (ESCs) generated from Thy1-EGFP transgenic (TG) mice. In addition, 3D retinal organoids differentiated from human induced PSCs (iPSCs) were cultured for up to differentiation day (DD) 120, and RGCs were purified by immunopanning. RGC marker expressions were confirmed by immunostaining and reverse transcription-quantitative PCR. The purified RGCs were cultured, and neurite outgrowth was measured and analyzed using an IncuCyte Zoom system. RESULTS.Mouse RGCs purified from Thy1-EGFP TG mouse retinas and the ESC-derived 3D retinas could be maintained for approximately 2 to 3 weeks, expressing the markers BRN3B and SMI-312. Purified RGCs from human iPSC-derived retinal organoids expressed RGC markers and could be maintained for up to 4 weeks. The RGCs collected at DD 90 to 110 extended longer neurites than those collected at younger stages.CONCLUSIONS. We successfully purified RGCs from mouse and human PSC-derived 3D retinal organoids cultured for approximately 120 days. RGCs from older retinal organoids would be useful for neurite tracking. This method would be effective not only for studying the pathology of human RGC diseases but also for therapeutic drug studies and RGC transplantation.

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Molecular codes for cell type specification in Brn3 retinal ganglion cells

Cited by 64 publications

References 89 publications

Essential Roles of Tbr1 in the Formation and Maintenance of the Orientation-Selective J-RGCs and a Group of OFF-Sustained RGCs in Mouse

Essential Roles of Tbr1 in the Formation and Maintenance of the Orientation-Selective J-RGCs and a Group of OFF-Sustained RGCs in Mouse

Differential expression and subcellular localization of Copines in mouse retina

Culture Systems of Dissociated Mouse and Human Pluripotent Stem Cell–Derived Retinal Ganglion Cells Purified by Two-Step Immunopanning

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