Retinal ganglion cell differentiation in cultured mouse retinal explants.
Methods
; 28(4): 448-56, 2002 Dec.
Article
en En
| MEDLINE
| ID: mdl-12507463
ABSTRACT
The availability of genetically engineered mice harboring specific mutations in genes affecting one or more retinal cell types affords new opportunities for investigating the genetic regulatory mechanisms of vertebrate retina formation. When identifying critical regulatory genes involved in retina development it is often advantageous to complement in vivo analysis with in vitro characterization. In particular, by combining classical techniques of retinal explant culturing with gene transfer procedures relying on herpes simple virus (HSV) amplicon vectors, gain-of-function analysis with genes of interest can be performed quickly and efficiently. Here, details are provided for isolating and culturing explants containing retinal progenitor cells and for infecting the explants with HSV expression vectors that perturb or rescue retinal ganglion cells, the first cell type to differentiate in the retina. In addition, the availability of sensitive techniques to monitor gene expression, including detection of reporter gene expression using antibodies and detection of endogenous marker gene expression using quantitative RT-PCR, provides an effective means for comparing wild-type and mutant retinas from genetically engineered mice.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Células Ganglionares de la Retina
/
Técnicas de Transferencia de Gen
/
Técnicas de Cultivo de Célula
Límite:
Animals
Idioma:
En
Revista:
Methods
Asunto de la revista:
BIOQUIMICA
Año:
2002
Tipo del documento:
Article
País de afiliación:
Estados Unidos