Differential mitochondrial DNA and gene expression in inherited retinal dysplasia in miniature Schnauzer dogs

Investigative Ophthalmology & Visual Science
Greg D AppleyardBruce H Grahn

Abstract

To investigate the molecular basis of inherited retinal dysplasia in miniature Schnauzers. Retina and retinal pigment epithelial tissues were collected from canine subjects at the age of 3 weeks. Total RNA isolated from these tissues was reverse transcribed to make representative cDNA pools that were compared for differences in gene expression by using a subtractive hybridization technique referred to as representational difference analysis (RDA). Expression differences identified by RDA were confirmed and quantified by real-time reverse-transcription PCR. Mitochondrial morphology from leukocytes and skeletal muscle of normal and affected miniature Schnauzers was examined by transmission electron microscopy. RDA screening of retinal pigment epithelial cDNA identified differences in mRNA transcript coding for two mitochondrial (mt) proteins--cytochrome oxidase subunit 1 and NADH dehydrogenase subunit 6--in affected dogs. Contrary to expectations, these identified sequences did not contain mutations. Based on the implication of mt-DNA-encoded proteins by the RDA experiments we used real-time PCR to compare the relative amounts of mt-DNA template in white blood cells from normal and affected dogs. White blood cells of affected dog...Continue Reading

Citations

May 1, 2016·Cell Calcium·Roberta A Gottlieb, Daniel Bernstein
Sep 26, 2013·BioMed Research International·Subhadip ChakrabortyKunal Ray
Sep 14, 2019·The Cochrane Database of Systematic Reviews·Cintia S de PaivaEsen K Akpek
Apr 13, 2007·Journal of the Optical Society of America. A, Optics, Image Science, and Vision·Yuhua Zhang, Austin Roorda

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