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(Investigative Ophthalmology and Visual Science. 2005;46:3000-3007.)
© 2005 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.05-0043
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Identification of P2Y Receptor Subtypes in Human Müller Glial Cells by Physiology, Single Cell RT-PCR, and Immunohistochemistry

Julia E. Fries,1 Iwona M. Goczalik,2 Thomas H. Wheeler-Schilling,1 Konrad Kohler,1 Elke Guenther,1,3 Sebastian Wolf,4,5 Peter Wiedemann,4 Andreas Bringmann,4 Andreas Reichenbach,2 Mike Francke,2 and Thomas Pannicke2

1From the Forschungsstelle für Experimentelle Ophthalmologie, Universitätsaugenklinik, Universität Tübingen, Tübingen, Germany; 2Paul-Flechsig-Institut für Hirnforschung und 4Klinik und Poliklinik für Augenheilkunde, Universität Leipzig, Leipzig, Germany; 3Naturwissenschaftliches und Medizinisches Institut (NMI) an der Universität Tübingen, Reutlingen, Germany.

PURPOSE. Retinal Müller glial cells are known to express metabotropic P2Y receptors. The present study was conducted to identify certain subtypes of P2Y receptors in human Müller cells.

METHODS. The patch-clamp technique was used to measure increases of Ca2+-dependent K+ currents mediated by the activation of P2Y receptors in freshly isolated human Müller cells. Several P2 agonists were used. Subsequently, the cells were harvested into the patch pipette and a single cell RT-PCR was performed. Moreover, retinal tissue from organ donors was used for immunohistochemistry.

RESULTS. The electrophysiological data were consistent with the expression of P2Y1, P2Y2, P2Y4, and P2Y6 receptor subtypes. RT-PCR revealed that mRNA for all these subtypes was present in Müller cells. However, the incidence of P2Y2 receptor mRNA was significantly lower than that of the other subtypes. Immunoreactivity for all four subtypes was found in retinal tissue, partly colocalized with immunoreactivity for vimentin.

CONCLUSIONS. The presented data obtained by different techniques revealed that human Müller cells express P2Y1, P2Y2, P2Y4, and P2Y6 receptors. The specific roles of these receptor subtypes in retinal physiology and/or pathophysiology remain to be investigated in future studies.




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