Functional characterization of mouse RDH11 as a retinol dehydrogenase involved in dark adaptation in vivo.
We previously cloned mouse RDH11 (mRDH11) as a gene regulated by the transcription factor sterol regulatory element-binding proteins and showed that it is a retinol dehydrogenase expressed in non-ocular tissues such as the liver and testis and in the retina (Kasus-Jacobi, A., Ou, J., Bashmakov, Y. K., Shelton, J. M., Richardson, J. A., Goldstein, J. L., and Brown, M. S. (2003) J. Biol. Chem. 278, 32380-32389). It was proposed to function in the recycling of the visual chromophore 11-cis-retinal after photoisomerization by a bleaching light, a pathway referred to as the visual cycle. In this work, we describe our studies on the ocular function of mRDH11. We created a knockout mouse by replacing the mrdh11 coding sequence with the lacZ reporter gene for expression profiling. 5-Bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) staining demonstrated active transcription of this gene in photoreceptor cells. We show by immunoblot analysis that mRDH11 is associated with retinal membranes purified from a non-outer segment fraction of the retina. No obvious retinal defect was found during development and aging of RDH11-deficient mice. The functional consequences of mRDH11 disruption were investigated by electroretinography. Dark adaptation was delayed by a factor of 2.5-3 compared with wild-type mice. However, the kinetics of 11-cis-retinal recycling during dark adaptation was not affected, suggesting that mRDH11 is not involved in the visual cycle. We propose that mRDH11 disruption affects retinoid metabolism in photoreceptor inner segments and delays the kinetics of dark adaptation through modulation of calcium homeostasis.
Kasus-Jacobi
Anne
A
Departmens of Molecular Genetics, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA. anne-kasus-jacobi@ouhsc.edu
Ou
Jiafu
J
Birch
David G
DG
Locke
Kirsten G
KG
Shelton
John M
JM
Richardson
James A
JA
Murphy
Andrew J
AJ
Valenzuela
David M
DM
Yancopoulos
George D
GD
Edwards
Albert O
AO
D000375:Aging; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis; D003623:Dark Adaptation / Q000502:physiology*; D004351:Drug Resistance / Q000235:genetics; D004596:Electroretinography; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D006706:Homeostasis; D015151:Immunoblotting; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009355:Neomycin; D010088:Oxidoreductases / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis; D012160:Retina / Q000201:enzymology; D017948:Retinal Rod Photoreceptor Cells / Q000201:enzymology; D012176:Retinoids / Q000032:analysis / Q000378:metabolism; D013737:Testis / Q000201:enzymology; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D012333:RNA, Messenger; D012176:Retinoids; D004247:DNA; D010088:Oxidoreductases; C486028:Rdh11 protein, mouse; D001616:beta-Galactosidase; D009355:Neomycin; D002118:Calcium