Figure 7. Restoration of GCAP2 expression in GCAPs/ retinas.

Figure 7

Restoration of GCAP2 expression in GCAPs/ retinas. A, Construct used to generate transgenic mice expressing bovine GCAP2 in the retina. A 5.7kb fusion gene was obtained by assembling the 4.4kb mouse opsin promoter, the 0.7kb bovine GCAP2 cDNA, and the mouse protamine 1 polyadenylation signal. B, Estimation of the transgene expression level by immunoblot. Fractions corresponding to 1/40 and 1/80 of a retina from mice expressing both the endogenous mGCAP2 and heterologous bGCAP2 (3 aa different in size) were separated by SDS/PAGE (lanes 1 and 2) and compared to fixed amounts of bovine and murine recombinant Histagged GCAP2 (120 to 15 ng, twofold dilutions). Proteins were detected with polyclonal Ab p24DN, raised against the bovine form of GCAP2.5 Heterologous GCAP2 expression was estimated to be 2fold higher than the endogenous GCAP2. C, Immunolocalization of bGCAP2 in the GCAPs/GCAP2+ retina. D, GC activity in retinal homogenates from wildtype, GCAPs/ and GCAPs/GCAP2+ mice at low (< 50 nM) or high (2 mM) free [Ca2+]. Values are the average of three independent experiments. Error bars represent the standard deviation. Reprinted from: Mendez A, Burns ME, Sokal I et al. Role of guanylate cyclaseactivating proteins (GCAPs) in setting the flash sensitivity of rod photoreceptors. Proc Natl Acad Sci USA 2001; 98: 99489953. “Copyright (2001) National Academy of Sciences, USA.”

From: Mouse Models to Study GCAP Functions in Intact Photoreceptors

Cover of Madame Curie Bioscience Database
Madame Curie Bioscience Database [Internet].
Austin (TX): Landes Bioscience; 2000-2013.
Copyright © 2000-2013, Landes Bioscience.

NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.