10 páginas, 5 figuras.
[Background] The neural retina is a highly structured tissue of the central nervous system that is
formed by seven different cell types that are arranged in layers. Despite much effort, the genetic
mechanisms that underlie retinal development are still poorly understood. In recent years, largescale
genomic analyses have identified candidate genes that may play a role in retinal neurogenesis,
axon guidance and other key processes during the development of the visual system. Thus, new
and rapid techniques are now required to carry out high-throughput analyses of all these candidate
genes in mammals. Gene delivery techniques have been described to express exogenous proteins
in the retina of newborn mice but these approaches do not efficiently introduce genes into the only
retinal cell type that transmits visual information to the brain, the retinal ganglion cells (RGCs).
[Results] Here we show that RGCs can be targeted for gene expression by in utero
electroporation of the eye of mouse embryos. Accordingly, using this technique we have
monitored the morphology of electroporated RGCs expressing reporter genes at different
developmental stages, as well as their projection to higher visual targets.
[Conclusion] Our method to deliver ectopic genes into mouse embryonic retinas enables us to
follow the course of the entire retinofugal pathway by visualizing RGC bodies and axons. Thus, this
technique will permit to perform functional studies in vivo focusing on neurogenesis, axon guidance,
axon projection patterning or neural connectivity in mammals.
This work was supported by grants to E. H. from
Human Frontiers Science Program (CDA-0023) and from the Spanish Government
(BFU-2004-0058). E.H. is a Ramón y Cajal Investigator from the
Consejo Superior de Investigaciones Científicas (CSIC).
Peer reviewed