Retinal degeneration in mice lacking the γ subunit of the rod cGMP phosphodiesterase

Science

Volumn 272, Issue 5264, 1996, Pages 1026-1029

  Tsang, Stephen H ^a   Gouras, Peter ^b   Yamashita, Clyde K ^c   Kjeldbye, Hild ^b   Fisher, John ^a,d   Farber, Debora B ^c   Goff, Stephen P ^a  

^a Columbia University ^*  (United States)

^b New York Presbyterian Hospital   (United States)

^c JULES STEIN EYE INSTITUTE   (United States)

^d REGENERON PHARMACEUTICALS INC   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CYCLIC GMP PHOSPHODIESTERASE;

ANIMAL MODEL; ARTICLE; ENZYME LOCALIZATION; ENZYME SUBUNIT; GENE MUTATION; GENE TARGETING; MOUSE; NONHUMAN; PHOTOTRANSDUCTION; PRIORITY JOURNAL; RETINA DEGENERATION; RETINITIS PIGMENTOSA;

EID: 0029740113     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.272.5264.1026     Document Type: Article

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41. The targeted mutation at the Pdeg locus was introduced into the germ line of chimeric mice by injection of the targeted ES clones into C57BL/6 or MF1 blastocysts that were surgically reimplanted into the uteri of foster mothers [V. E. Papaioannou and R. Johnson, in Gene Targeting, A. L. Joyner, Ed. (Oxford Univ. Press, New York, 1993), pp. 107-146; A. Bradley, in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, E. J. Robertson, Ed. (IRL, Oxford, 1987), pp. 113-151]. Thirty-six chimeras were bom and mated with C57BL/6 or MF1 mice to test tor germline competency of the targeted clones; 29 of these chimeric mice transmitted the mutant allele. ES cell-derived progeny were identified by coat color and screened for the presence of the targeted allele by Southern blot or polymerase chain reaction (PCR) obtain 0.1 -s pulses that were repeated every 5 to 10 s at high stimulus intensities. The maximal responses at 2 to 3 weeks after birth were 600 to 800 μV from normal mice but only 20 to 30 μV from mutants.
42. S. H. Tsang et al., unpublished data.
43. Retinas were fixed by vascular perfusion of the mice with a solution of 3% glutaraldehyde and 1.5% paraformaldehyde in phosphate-buffered saline at pH 7.2. Epon-embedded blocks were prepared as described [P , Gouras, J. Du, H. Kjeldbye, S. Yamamoto, D. Zack, Invest. Ophthalmol. Vis. Sci. 35, 3145 (1994)], Single sections of the retina extending from the ora serrata superiorally to the ora serrata inferiorally, including the optic nerve, were examined by light microscopy; selected areas were also studied by electron microscopy.
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50. We thank R. Axel, F. Costantini, B. K.-K. Fung, J. L. German III, P. Mombaerts, A. Nagy, V. E, Papaioannou, J. Rossant, A. Tomlinson, D. Warburton, and members of their laboratories for sharing ideas, cell lines, antisera, and equipment, and for critically reading the manuscript; E. Carlson, C.-S. Lin, M. Mendelsohn, and F. Wang for guidance and advice; and members of the Farber, Goff, and Gouras laboratories for support, especially W. Li. S.H.T. is supported by Medical Scientist Training Program fellowship T32 GM 073667-14. Supported by NIH grants EY08285 (D.B.F.) and by grants from the George Gund Foundation and the Foundation Fighting Blindness (D.B.F.). D.B.F. is the recipient of a Senior Scientific Investigator Award from Research to Prevent Blindness. S.P.G. is an Investigator of the Howard Hughes Medical Institute.