Modulation of the expression of the transcription factor Max in rat retinal ganglion cells by a recombinant adeno-associated viral vector.
Exclusion of the transcription factor Max from the nucleus of retinal ganglion cells is an early, caspase-independent event of programmed cell death following damage to the optic axons. To test whether the loss of nuclear Max leads to a reduction in neuroprotection, we developed a procedure to overexpress Max protein in rat retinal tissue in vivo. A recombinant adeno-associated viral vector (rAAV) containing the max gene was constructed, and its efficiency was confirmed by transduction of HEK-293 cells. Retinal ganglion cells were accessed in vivo through intravitreal injections of the vector in rats. Overexpression of Max in ganglion cells was detected by immunohistochemistry at 2 weeks following rAAV injection. In retinal explants, the preparation of which causes damage to the optic axons, Max immunoreactivity was increased after 30 h in vitro, and correlated with the preservation of a healthy morphology in ganglion cells. The data show that the rAAV vector efficiently expresses Max in mammalian retinal ganglion cells, and support the hypothesis that the Max protein plays a protective role for retinal neurons.
Petrs-Silva
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Instituto de Biofísica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brasil. petrs@biof.ufrj.br
Chiodo
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V
Chiarini
L B
LB
Hauswirth
W W
WW
Linden
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D000818:Animals; D000831:Animals, Newborn; D001369:Axons; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D007150:Immunohistochemistry; D009410:Nerve Degeneration / Q000378:metabolism; D010321:Parvoviridae; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000378:metabolism* / Q000473:pathology
D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; C499329:Max protein, rat; D011994:Recombinant Proteins