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Localization of caveolin-1 and c-src in mature and differentiating photoreceptors: raft proteins co-distribute with rhodopsin during development

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Abstract

Numerous biochemical and morphological studies have provided insight into the distribution pattern of caveolin-1 and the presence of membrane rafts in the vertebrate retina. To date however, studies have not addressed the localization profile of raft specific proteins during development. Therefore the purpose of our studies was to follow the localization pattern of caveolin-1, phospho-caveolin-1 and c-src in the developing retina and compare it to that observed in adults. Specific antibodies were used to visualize the distribution of caveolin-1, c-src, a kinase phosphorylating caveolin-1, and phospho-caveolin-1. The labeling pattern of this scaffolded complex was compared to those of rhodopsin and rhodopsin kinase. Samples were analyzed at various time points during postnatal development and compared to adult retinas. The immunocytochemical studies were complemented with immunoblots and immunoprecipitation studies. In the mature retina caveolin-1 and c-src localized mainly to the cell body and IS of photoreceptors, with only very weakly labeled OS. In contrast, phospho-caveolin-1 was only detectable in the OS of photoreceptors. During development we followed the expression and distribution profile of these proteins in a temporal sequence with special attention to the period when OS formation is most robust. Double labeling immunocytochemistry and immunoprecipitation showed rhodopsin to colocalize and co-immunoprecipitate with caveolin-1 and c-src. Individual punctate structures between the outer limiting membrane and the outer plexiform layer were seen at P10 to be labeled by both rhodopsin and caveolin-1 as well as by rhodopsin and c-src, respectively. These studies suggest that membrane raft specific proteins are co-distributed during development, thereby pointing to a role for such complexes in OS formation. In addition, the presence of small punctate structures containing caveolin-1, c-src and rhodopsin raise the possibility that these proteins may transport together to OS during development and that caveolin-1 exists predominantly in a phosphorylated form in the OS.

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Acknowledgments

We would like to thank Dr. Pál Röhlich for critically reading the manuscript and Dr. Gustavo D. Aguirre for his beneficial advices about the manuscript. We are grateful to Dr. Krzysztof Palczewski for the donation of the antibody against rhodopsin kinase, to Dr. Robert Molday for the donation of the RDS antibody, to Nándor Müllner for the immunoprecipitation and also to Katalin Lőcsey and Margit Kutasi for their help in the sample preparation for electron microscopy. The work was supported by Hungarian OTKA Grant, #73000 (Ágoston Szél), EY10420 and EY18705 (Kathleen Boesze-Battaglia).

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Authors and Affiliations

  1. Department of Human Morphology and Developmental Biology, Semmelweis University Budapest, Tűzoltó u. 58, Budapest, Hungary

    Ágnes I. Berta, Attila Magyar, Ágoston Szél & Anna L. Kiss

  2. Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, 240 South 40th Street, Philadelphia, PA, 19104, USA

    Kathleen Boesze-Battaglia

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  1. Ágnes I. Berta
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  2. Kathleen Boesze-Battaglia
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  3. Attila Magyar
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  4. Ágoston Szél
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  5. Anna L. Kiss
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Correspondence to Anna L. Kiss.

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Berta, Á.I., Boesze-Battaglia, K., Magyar, A. et al. Localization of caveolin-1 and c-src in mature and differentiating photoreceptors: raft proteins co-distribute with rhodopsin during development. J Mol Hist 42, 523–533 (2011). https://doi.org/10.1007/s10735-011-9360-4

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  • DOI: https://doi.org/10.1007/s10735-011-9360-4

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