Abstract
Abstract: :
Purpose: Fovin is a member of the Tweety gene family whose protein function is unknown. Northern blot analysis shows that fovin is highly expressed in human and mouse retinas. The goal of this study was to identify fovin’s cellular location in the mouse retina in order gain insight into fovin’s function Methods: Polyclonal sera against fovin synthetic peptides were produced in rabbits. One of these antibodies, DEBRH1–80, recognizes a single band on Western blots of 59 kDA in mouse retina protein extracts. Immunocytochemistry was performed on frozen tissue sections of mouse eyes using antibody DEBRH1–80 and a Cy3 labeled secondary antibody. Cell nuclei were identified by DAPI staining. In situ hybridization was also performed on frozen tissue sections of mouse eyes. Fovin sense and antisense digoxigenin–labeled RNA probes were synthesized by in vitro transcription and hybridized to tissue sections of mouse eyes. Results: When viewed by immunofluorescence microscopy, ganglion cells react with the fovin antibody but not with the preimmune control serum. The in situ hybridization experiments confirm these results. The fovin antisense but not the sense probe hybridized to ganglion cells. Conclusions: These experiments show that mouse retinal ganglion cells express fovin mRNA and synthesize fovin protein. These results are not unexpected because the fovin cDNA clone was originally isolated from a human fovea primary cDNA library. The fovea contains a high number of ganglion cells. Knowledge of fovin’s cellular localization will guide us to understanding fovin’s role in retinal ganglion cell biology.
Keywords: ganglion cells • immunohistochemistry • in situ hybridization