Differential expression of GAD65 and GAD67 during the development of the rat retina

Dkhissi, Ouria; Julien, Jean‐François; Wasowicz, Marguerite Y; Dalil-Thiney, Najet; Nguyen‐Legros, Jeanine; Versaux‐Botteri, Claudine

doi:10.1016/s0006-8993(01)03022-0

Cited by 32 publications

(35 citation statements)

References 29 publications

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“…Staining in the retina was as previously reported and was monitored as control. In all cases, it was identical to the previously reported pattern (Haverkamp and Wassle, 2000;Dkhissi et al, 2001;de Melo et al, 2003). Based on the reverse transcriptase-polymerase chain reaction and Western blot results, most, if not all GAD67-related immunoreactivity during embryonic stages is due to EGAD (GAD25 and GAD44); only adult GAD67 is expressed beyond P7.…”

Section: Dlx2 Expression Labels Both Epithelium and Fiber Cells Of Thsupporting

confidence: 87%

GAD isoforms exhibit distinct spatiotemporal expression patterns in the developing mouse lens: Correlation with Dlx2 and Dlx5

Kwakowsky

Schwirtlich

Zhang

et al. 2007

Developmental Dynamics

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Gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter of the adult nervous system and its biosynthetic enzyme glutamic acid decarboxylase (GAD) are abundantly expressed in the embryonic nervous system and are involved in the modulation of cell proliferation, migration, and differentiation. Here we describe for the first time the expression of GABA and embryonic and adult GAD isoforms in the developing mouse lens. We show that the GAD isoforms are sequentially induced with specific spatiotemporal profiles: GAD65 and embryonic GAD isoforms prevail in primary fibers, while GAD67 is the predominant GAD expressed in the postnatal secondary fibers. This pattern correlates well with the expression of Dlx2 and Dlx5, known as upstream regulators of GAD. GABA and GAD are most abundant at the tips of elongating fibers and are absent from organelle-free cells, suggesting their involvement is primarily in shaping of the cytoskeleton during fiber elongation stages. Developmental Dynamics 236: 3532-3544, 2007.

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Section: Dlx2 Expression Labels Both Epithelium and Fiber Cells Of Thsupporting

confidence: 87%

GAD isoforms exhibit distinct spatiotemporal expression patterns in the developing mouse lens: Correlation with Dlx2 and Dlx5

Kwakowsky

Schwirtlich

Zhang

et al. 2007

Developmental Dynamics

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“…Our RT-qPCR showed that the purified cells expressed RGC-specific genes, and no other celltype markers except GAD1 were detected. Because GAD1 is expressed not only in developing RGCs but also in amacrine cells, 36 our results indicate that these purified RGCs were at early stages of growth or development.…”

Section: Discussionmentioning

confidence: 66%

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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“…GAD, which is a rate-limiting enzyme in GABA synthesis from glutamic acid, is a marker of GABAergic neurons (12). In retina, it was reported that GAD is expressed in GABAergic amacrine cells and in a subset of horizontal cells (5,34). As shown in Fig.…”

Section: Characterization Of Anti-kir51 Antibody and Localization Ofmentioning

confidence: 95%

Differential expression and distribution of Kir5.1 and Kir4.1 inwardly rectifying K⁺ channels in retina

Ishii

Fujita²,

Iwai³

et al. 2003

American Journal of Physiology-Cell Physiology

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Kir5.1 is an inwardly rectifying K+ channel subunit whose functional role has not been fully elucidated. Expression and distribution of Kir5.1 in retina were examined with a specific polyclonal antibody. Kir5.1 immunoreactivity was detected in glial Müller cells and in some retinal neurons. In the Kir5.1-positive neurons the expression of glutamic acid decarboxylase (GAD65) was detected, suggesting that they may be GABAergic-amacrine cells. In Müller cells, spots of Kir5.1 immunoreactivity distributed diffusely at the cell body and in the distal portions, where Kir4.1 immunoreactivity largely overlapped. In addition, Kir4.1 immunoreactivity without Kir5.1 was strongly concentrated at the endfoot of Müller cells facing the vitreous surface or in the processes surrounding vessels. The immunoprecipitant obtained from retina with anti-Kir4.1 antibody contained Kir5.1. These results suggest that heterotetrameric Kir4.1/Kir5.1 channels may exist in the cell body and distal portion of Müller cells, whereas homomeric Kir4.1 channels are clustered in the endfeet and surrounding vessels. It is possible that homomeric Kir4.1 and heteromeric Kir4.1/Kir5.1 channels play different functional roles in the K+-buffering action of Müller cells.

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Differential expression of GAD65 and GAD67 during the development of the rat retina

Cited by 32 publications

References 29 publications

GAD isoforms exhibit distinct spatiotemporal expression patterns in the developing mouse lens: Correlation with Dlx2 and Dlx5

GAD isoforms exhibit distinct spatiotemporal expression patterns in the developing mouse lens: Correlation with Dlx2 and Dlx5

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

Differential expression and distribution of Kir5.1 and Kir4.1 inwardly rectifying K⁺ channels in retina

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Differential expression of GAD65 and GAD67 during the development of the rat retina

Cited by 32 publications

References 29 publications

GAD isoforms exhibit distinct spatiotemporal expression patterns in the developing mouse lens: Correlation with Dlx2 and Dlx5

GAD isoforms exhibit distinct spatiotemporal expression patterns in the developing mouse lens: Correlation with Dlx2 and Dlx5

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

Differential expression and distribution of Kir5.1 and Kir4.1 inwardly rectifying K+ channels in retina

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Differential expression and distribution of Kir5.1 and Kir4.1 inwardly rectifying K⁺ channels in retina