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Immunohistochemical and Calcium Imaging Methods in Wholemount Rat Retina
Published on: October 13, 2014
Stress-induced changes in neuronal Aquaporin-9 (AQP9) in a retinal ganglion cell-line
Adnan Dibas1, Ming-Hui Yang, Joseph Bobich
1Department of Pharmacology & Neuroscience, University of North Texas Health Science Center at Fort Worth, TX 76107, USA. adibas@hsc.unt.edu
Abstract:
The water channel, Aquaporin-9 (AQP9) is enriched in selected neuronal populations and is unique its ability to act as a lactate-glycerol channel supplying neurons with alternative fuel under ischaemic conditions. AQP9 was detected in RGC-5 cells, a retinal ganglion cell-line, primary RGCs, and retina by Western blotting, real-time PCR (RT-PCR) and immunohistochemistry. RGC-5 cells subjected to a hypotonic stress increased their cell volume that was blocked by known inhibitor of AQP9 (phloretin (40 microM)). RGC-5 cells subjected to hypoxia, showed an up-regulation in AQP9 expression as judged by Western blotting and RT-PCR. Similarly, hypotonic shock (50%) increased AQP9 expression as determined by RT-PCR. AQP9 is involved in energy balance as a glycerol-lactate channel and also appears to regulate cell volume in retinal ganglion neurons. This water channel may play a key role in retinal ganglion pathology.

