Abstract
Mutations in rod opsin, the visual pigment protein of rod photoreceptors, account for approximately 15% of all inherited human retinal degenerations. However, the physiological and molecular events underlying the disease process are not well understood. One approach to this question has been to study transgenic mice expressing opsin genes containing defined mutations. A caveat of this approach is that even the overexpression of normal opsin leads to photoreceptor cell degeneration. To overcome the problem, we have reduced or eliminated endogenous rod opsin content by targeted gene disruption. Retinas in mice lacking both opsin alleles initially developed normally, except that rod outer segments failed to form. Within months of birth, photoreceptor cells degenerated completely. Retinas from mice with a single copy of the opsin gene developed normally, and rods elaborated outer segments of normal size but with half the normal complement of rhodopsin. Photoreceptor cells in these retinas also degenerated but did so over a much slower time course. Physiological and biochemical experiments showed that rods from mice with a single opsin gene were approximately 50% less sensitive to light, had accelerated flash-response kinetics, and contained approximately 50% more phosducin than wild-type controls.
MeSH Terms
Animals
Electrophysiology
Eye Proteins/genetics
Gene Targeting/methods
Light
Mice
Mice, Knockout
Microspectrophotometry
Photoreceptor Cells, Vertebrate/physiology
Retinal Rod Photoreceptor Cells/physiology
Rhodopsin/genetics
Rod Opsins/genetics
Vision, Ocular/genetics
Chemicals
Eye Proteins
Rod Opsins
Rhodopsin
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Lem J
New England Medical Center, Tufts University School of Medicine, Boston, MA 02111, USA.
[email protected]
Krasnoperova N V
Calvert P D
Kosaras B
Cameron D A
Nicolò M
Makino C L
Sidman R L
References (28)
28 references, click to expand
-
A null mutation in the rhodopsin gene causes rod photoreceptor dysfunction and autosomal recessive retinitis pigmentosa.
Nat Genet. 1992 Jun;1(3):209-13
PMID: 1303237
-
Phosducin is a protein kinase A-regulated G-protein regulator.
Nature. 1992 Jul 2;358(6381):73-6
PMID: 1319556
-
Transgenic mice with a rhodopsin mutation (Pro23His): a mouse model of autosomal dominant retinitis pigmentosa.
Neuron. 1992 Nov;9(5):815-30
PMID: 1418997
-
A third form of the G protein beta subunit. 1. Immunochemical identification and localization to cone photoreceptors.
J Biol Chem. 1992 Dec 5;267(34):24776-81
PMID: 1447215
-
Targeted mutation of the DNA methyltransferase gene results in embryonic lethality.
Cell. 1992 Jun 12;69(6):915-26
PMID: 1606615
-
Recoverin: a calcium sensitive activator of retinal rod guanylate cyclase.
Science. 1991 Feb 22;251(4996):915-8
PMID: 1672047
-
Anti-rhodopsin monoclonal antibodies of defined specificity: characterization and application.
Vision Res. 1991;31(1):17-31
PMID: 2006550
-
Interactions between the subunits of transducin and cyclic GMP phosphodiesterase in Rana catesbiana rod photoreceptors.
J Biol Chem. 1990 Jul 15;265(20):11539-48
PMID: 2164007
-
Isolation and characterization of cGMP phosphodiesterase from bovine rod outer segments.
J Biol Chem. 1979 Nov 25;254(22):11669-77
PMID: 227876
-
The photoreceptor-specific 33 kDa phosphoprotein of mammalian retina: generation of monospecific antibodies and localization by immunocytochemistry.
Exp Eye Res. 1988 Jun;46(6):829-40
PMID: 2461862
-
A new method of non-enzymatic dissociation of the Bufo retina.
J Neurosci Methods. 1986 Feb;15(4):301-6
PMID: 3083159
-
A novel complex from bovine visual cells of a 33,000-dalton phosphoprotein with beta- and gamma-transducin: purification and subunit structure.
Biochemistry. 1987 Jun 30;26(13):3983-90
PMID: 3477288
-
The visual pigments of rods and cones in the rhesus monkey, Macaca mulatta.
J Physiol. 1978 Jan;274:329-48
PMID: 415133
-
The subcellular localization of rat photoreceptor-specific antigens.
J Neurocytol. 1983 Oct;12(5):785-803
PMID: 6358424
-
A rhodopsin gene mutation responsible for autosomal dominant retinitis pigmentosa results in a protein that is defective in localization to the photoreceptor outer segment.
J Neurosci. 1994 Oct;14(10):5818-33
PMID: 7523628
-
The N terminus of phosducin is involved in binding of beta gamma subunits of G protein.
Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2086-90
PMID: 7534410
-
Constitutive activation of phototransduction by K296E opsin is not a cause of photoreceptor degeneration.
Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3551-5
PMID: 7724596
-
Mechanisms of rhodopsin inactivation in vivo as revealed by a COOH-terminal truncation mutant.
Science. 1995 Jan 20;267(5196):374-7
PMID: 7824934
-
The phosphorylation state of phosducin determines its ability to block transducin subunit interactions and inhibit transducin binding to activated rhodopsin.
J Biol Chem. 1994 Sep 30;269(39):24050-7
PMID: 7929057
-
Absorption spectra and linear dichroism of some amphibian photoreceptors.
J Gen Physiol. 1975 Sep;66(3):357-82
PMID: 808586
-
Cellular interactions implicated in the mechanism of photoreceptor degeneration in transgenic mice expressing a mutant rhodopsin gene.
Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8484-8
PMID: 8378322
-
Temperature dependence of the light response in rat rods.
J Physiol. 1993 Mar;462:465-81
PMID: 8392571
-
Simulation of human autosomal dominant retinitis pigmentosa in transgenic mice expressing a mutated murine opsin gene.
Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5499-503
PMID: 8516292
-
Rhodopsin plays an essential structural role in Drosophila photoreceptor development.
Development. 1995 Dec;121(12):4359-70
PMID: 8575336
-
Transgenic mice carrying the dominant rhodopsin mutation P347S: evidence for defective vectorial transport of rhodopsin to the outer segments.
Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):14176-81
PMID: 8943080
-
A splice variant of arrestin from human retina.
Exp Eye Res. 1996 Jun;62(6):585-92
PMID: 8983940
-
Visual pigment assignments in regenerated retina.
J Neurosci. 1997 Feb 1;17(3):917-23
PMID: 8994046
-
Retinopathy induced in mice by targeted disruption of the rhodopsin gene.
Nat Genet. 1997 Feb;15(2):216-9
PMID: 9020854