Hostname: page-component-cd9895bd7-p9bg8 Total loading time: 0 Render date: 2024-12-25T05:52:21.272Z Has data issue: false hasContentIssue false
  • English
  • Français

Reporter gene expression in cones in transgenic mice carrying bovine rhodopsin promoter/lacZ transgenes

Published online by Cambridge University Press:  02 June 2009

Peter Gouras ,
Hild Kjeldbye  and
Donald J. Zack
Peter Gouras
Affiliation:
Department of Ophthalmology, Harkness Eye Institute, Columbia University, New York
Hild Kjeldbye
Affiliation:
Department of Ophthalmology, Harkness Eye Institute, Columbia University, New York
Donald J. Zack*
Affiliation:
Departments of Ophthalmology, Molecular Biology and Genetics, and Neuroscience, Johns Hopkins University School of Medicine, Baltimore
*
Address correspondence to: Donald J. Zack, Wilmer Institute, Johns Hopkins University School of Medicine, 809 Maumenee, 600N. Wolfe street, Baltimore, MD 21205, USA
Get access Rights & Permissions [Opens in a new window]

Abstract

Rhodopsin gene expression has been used as a model system to study the mechanisms regulating photoreceptor gene expression. Previous transgenic experiments using rhodopsin promoter/lacZ fusion constructs identified some of the cis-acting DNA elements responsible for photoreceptor cell-specific expression. However, the issue of rod specificity vs. photoreceptor (rod and cone) specificity of the elements was not resolved. To address this issue, the specificity of reporter gene expression in the retinas of transgenic mice carrying bovine rhodopsin promoter/lacZ (ß-galactosidase) fusion genes was assessed using X-gal staining and electron microscopy. Two independent transgenic lines, one carrying a rhodopsin promoter fragment extending from −2174 to +70 base pairs (bp) relative to the messenger RNA start site and another line carrying a fragment from −222 to +70 bp, both showed reporter gene expression in cones as well as rods, although the level of staining appeared to be less in the cones than in the rods. These results demonstrate that the −2174 to +70 bp and −222 to +70 bp bovine rhodopsin promoter fragments are not rod-specific in transgenic mice and indicate that the existence of rod promoter mediated-expression in cones must be considered when interpreting results from transgenic experiments utilizing the rhodopsin promoter.

Keywords

Rhodopsin promoterRetinal gene expressionPhotoreceptorsConesTransgenic mice
Type
Short Communications
Information
Visual Neuroscience , Volume 11 , Issue 6 , November 1994 , pp. 1227 - 1231
Copyright
Copyright © Cambridge University Press 1994

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Bonnerot, C., Rocancourt, D., Briand, P., Grimber, G. & Nicolae, J. (1987). A beta galactosidase hybrid protein targeted to nuclei as a marker for developmental studies. Proceedings of the National Academy of Sciences of the U.S.A. 84, 6795–6799.Google Scholar
Dryja, T.P., McGee, T.L., Reichel, E., Hahn, L.B., Cowley, G.S., Yandell, D.W., Sandberg, M.A. & Berson, E.L. (1990). A point mutation of the rhodopsin gene in one form of retinitis pigmentosa. Nature 343, 364–366.Google Scholar
Du, J., Gouras, P., Kjeldbye, H., Kwun, R. & Lopez, R. (1992). Monitoring photoreceptor transplants with nuclear and cytoplasmic markers. Experimental Neurology 115, 79–86.CrossRefGoogle ScholarPubMed
Gouras, P., Du, J., Kjeldbye, H., Kwun, R., Lopez, R. & Zack, D.J. (1991). Transplanted photoreceptors identified in dystrophic mouse retina by a transgenic reporter gene. Investigative Ophthalmology and Visual Science 32, 3167–3174.Google Scholar
Gouras, P., Du, J., Kjeldbye, H., Yamamoto, S. & Zack, D.J. (1992). Reconstruction of degenerated rd mouse retina by transplantation of transgenic photoreceptors. Investigative Ophthalmology and Visual Science 33, 2579–2586.Google ScholarPubMed
Kumar, R. & Zack, D.J. (1994). Regulation of visual pigment gene expression. In Molecular Genetics of Ocular Disorders, ed. Wiggs, J., pp. 137–156. New York: Wiley Liss.Google Scholar
Lem, J., Applebury, M.L., Falk, J.D., Flannery, J.G. & Simon, M.I. (1991). Tissue-specific and developmental regulation of rod opsin chimeric genes in transgenic mice. Neuron 6, 201–210.CrossRefGoogle ScholarPubMed
Sung, C.-H., Davenport, C.M., Hennesey, J.C., Maumenee, I.H., Jacobson, S.G., Heckenlively, J.R., Nowakowski, R., Fishman, G., Gouras, P. & Nathans, J. (1991). Rhodopsin mutations in autosomal dominant retinitis pigmentosa. Proceedings of the National Academy of Sciences of the U.S.A. 88, 6481–6485.CrossRefGoogle ScholarPubMed
Woodford, B.J., Chen, J. & Simon, M.I. (1994). Expression of rhodopsin promoter transgene product in both rods and cones. Experimental Eye Research 58, 631–635.CrossRefGoogle ScholarPubMed
Zack, D.J. (1993 a). Analysis of retinal-specific gene expression in transgenic mice. In Methods in Neuroscience: Photoreceptor Cells, ed. Hargrave, P.A., pp. 331–341. Orlando, Florida: Academic Press.CrossRefGoogle Scholar
Zack, D.J. (1993 b). Ocular gene therapy. From fantasy to foreseeable reality. Archives of Ophthalmology 111, 1477–1479.CrossRefGoogle ScholarPubMed
Zack, D.J., Bennett, J., Wang, Y., Davenport, C., Klaunberg, B., Gearhart, J. & Nathans, J. (1991). Unusual topography of bovine rhodopsin promoter-lacZ fusion gene expression in transgenic mouse retinas. Neuron 6, 187–199.CrossRefGoogle ScholarPubMed