Rapid isolation of intact retinal astrocytes: a novel approach DOI Creative Commons
Paul Cullen, Arpan G. Mazumder, Daniel Sun

et al.

Acta Neuropathologica Communications, Journal Year: 2023, Volume and Issue: 11(1)

Published: Sept. 25, 2023

Astrocytes are a major category of glial support cell in the central nervous system and play variety essential roles both health disease. As our understanding diverse functions these cells improves, extent heterogeneity between astrocyte populations has emerged as key area research. Retinal astrocytes, which form direct cellular environment retinal ganglion somas axons, undergo reactive response human glaucoma animal models disease, yet their contributions to its pathology progression remain relatively unknown. This gap knowledge is largely function inadequate isolation techniques, driven part by sparseness similarities with more abundant Müller cells. Here, we present novel method isolating astrocytes enriching RNA, tested normal ocular hypertensive mice, common model experimental glaucoma. Our approach combines enzyme assisted microdissection selective ribosome immunoprecipitation using Ribotag method. rapid preserves morphology, resulting brief post-mortem interval minimizing loss RNA from distal regions Both require minimum specialized equipment or reagents, them conjunction able achieve > 100-fold enrichment RNA.

Language: Английский

Rapid isolation of intact retinal astrocytes: a novel approach DOI Creative Commons
Paul Cullen, Arpan G. Mazumder, Daniel Sun

et al.

Acta Neuropathologica Communications, Journal Year: 2023, Volume and Issue: 11(1)

Published: Sept. 25, 2023

Astrocytes are a major category of glial support cell in the central nervous system and play variety essential roles both health disease. As our understanding diverse functions these cells improves, extent heterogeneity between astrocyte populations has emerged as key area research. Retinal astrocytes, which form direct cellular environment retinal ganglion somas axons, undergo reactive response human glaucoma animal models disease, yet their contributions to its pathology progression remain relatively unknown. This gap knowledge is largely function inadequate isolation techniques, driven part by sparseness similarities with more abundant Müller cells. Here, we present novel method isolating astrocytes enriching RNA, tested normal ocular hypertensive mice, common model experimental glaucoma. Our approach combines enzyme assisted microdissection selective ribosome immunoprecipitation using Ribotag method. rapid preserves morphology, resulting brief post-mortem interval minimizing loss RNA from distal regions Both require minimum specialized equipment or reagents, them conjunction able achieve > 100-fold enrichment RNA.

Language: Английский

Citations

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