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B Bakall, F Aspgren, C Wadelius; Characterization of VMD2 Transcripts in Human and Transcription Levels in Mice . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3659.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To characterize VMD2 transcripts in human tissues and analyse expression levels of VMD2 in mice during development. Methods: RT-PCR was used to amplify alternative splice variants of VMD2 in human retina, brain and testis. Amplified fragments were sequenced. Balb/c mice eyes were isolated during different development stages and RNA was extracted. The relative expression of VMD2 in mice eyes was analyzed using quantitative RT-PCR (Taqman) with mouse GAPDH as a control. Results: RT-PCR of human tissues resulted in fragments of different sizes. Sequencing revealed a novel splice variant lacking exon 2. This variant was found predominantly in brain, but was also detected in retina and testis. The quantitative expression analysis (Taqman) of VMD2 in mice eyes during development showed that the expression is at its peak during embryonic stages and early postnatal life, decreasing with age. Conclusion: The identified novel splice variant without exon 2 is lacking the original ATG start codon. This has a potential to form an isoform of the protein bestrophin of 525 amino acids, by using an ATG start codon in exon 3, which is in frame with the known variant. The expression profile of VMD2 in mouse with the highest levels early in life indicates that it has relevance during retinal maturation.
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