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(Investigative Ophthalmology and Visual Science. 2006;47:348-356.)
© 2006 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.05-0858
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Developmental and Tissue Expression of Xenopus laevis RPGR

Xinhua Shu,1 Zhihong Zeng,2 Marion S. Eckmiller,3 Phillipe Gautier,1 Dafni Vlachantoni,1 Forbes D. C. Manson,4 Brian Tulloch,1 Colin Sharpe,5 Dariusz C. Gorecki,2 and Alan F. Wright1

1From the MRC Human Genetics Unit, Western General Hospital, Edinburgh, United Kingdom; the 2School of Pharmacy and Biomedical Sciences and the 5School of Biological Sciences, Institute of Biomedical and Biomolecular Sciences, University of Portsmouth, Portsmouth, United Kingdom; the 3C. and O. Vogt Brain Research Institute, Heinrich Heine University of Düsseldorf School of Medicine, Düsseldorf, Germany; and the 4Centre for Molecular Medicine, Academic Unit of Medical Genetics, Division of Human Development, The University of Manchester, Manchester, United Kingdom.

PURPOSE. The present study examined the developmental and tissue expression of the retinitis pigmentosa GTPase regulator (RPGR) gene in Xenopus laevis.

METHODS. The cDNA for X. laevis RPGR (XRPGR) was isolated from adult eye mRNA by reverse transcription–polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The deduced peptide sequence was aligned with RPGR orthologues. Gene expression was examined by whole-mount in situ hybridization and RT-PCR. The localization of XRPGR in X. laevis photoreceptor cells and XTC-2 cells was determined by immunostaining.

RESULTS. The XRPGRex1–19 isoform encodes a protein of 727 amino acids containing an RCC1 domain and a C-terminal isoprenylation anchorage motif. It shares 33% to 41% amino acid identity with human, mouse, and dog RPGR. The C-terminal exon of the alternatively spliced RPGRORF15 isoform is also conserved across species. XRPGR is expressed at the earliest stages of X. laevis development and persists into adulthood, where expression is highest in the eye. XRPGR is expressed in presumptive eye fields (stages 18 to 22), becoming largely restricted to the central retina (stages 28 to 40). XRPGR protein colocalizes with ß-tubulin at the X. laevis ciliary axoneme and with {gamma}-tubulin at centrosomes in XTC-2 cells.

CONCLUSIONS. XRPGR is widely expressed throughout development but shows highest expression after the appearance of the eye primordium and persists in the eye into adulthood. The data are consistent with XRPGR expression in a single microtubular organelle—the centriole or basal body and associated ciliary transitional zone found in modified sensory cilia of photoreceptors and motile cilia.






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