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Inhibition of Müller glial cell division blocks regeneration of the light‐damaged zebrafish retina
Author(s) -
Thummel Ryan,
Kassen Sean C.,
Montgomery Jacob E.,
Enright Jennifer M.,
Hyde David R.
Publication year - 2007
Publication title -
developmental neurobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.716
H-Index - 129
eISSN - 1932-846X
pISSN - 1932-8451
DOI - 10.1002/dneu.20596
Subject(s) - biology , muller glia , microbiology and biotechnology , morpholino , zebrafish , retina , gene knockdown , progenitor cell , proliferating cell nuclear antigen , outer nuclear layer , regeneration (biology) , electroporation , neuroglia , stem cell , cell growth , neuroscience , cell culture , central nervous system , biochemistry , genetics , gene
The adult zebrafish retina possesses a robust regenerative response. In the light‐damaged retina, Müller glial cell divisions precede regeneration of rod and cone photoreceptors. Neuronal progenitors, which arise from the Müller glia, continue to divide and use the Müller glial cell processes to migrate to the outer nuclear layer and replace the lost photoreceptors. We tested the necessity of Müller glial cell division for photoreceptor regeneration. As knockdown tools were unavailable for use in the adult zebrafish retina, we developed a method to conditionally inhibit the expression of specific proteins by in vivo electroporation of morpholinos. We determined that two separate morpholinos targeted against the proliferating cell nuclear antigen (PCNA) mRNA reduced PCNA protein levels. Furthermore, injection and in vivo electroporation of PCNA morpholinos immediately prior to starting intense light exposure inhibited both Müller glial cell proliferation and neuronal progenitor marker Pax6 expression. PCNA knockdown additionally resulted in decreased expression of glutamine synthetase in Müller glia and Müller glial cell death, while amacrine and ganglion cells were unaffected. Finally, histological and immunological methods showed that long‐term effects of PCNA knockdown resulted in decreased numbers of Müller glia and the failure to regenerate rod photoreceptors, short single cones, and long single cones. These data suggest that Müller glial cell division is necessary for proper photoreceptor regeneration in the light‐damaged zebrafish retina and are consistent with the Müller glia serving as the source of neuronal progenitor cells in regenerating teleost retinas. © 2007 Wiley Periodicals, Inc. Develop Neurobiol, 2008.