May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Expression of Transient Receptor Potential Channels in Retinal Arterioles
Author Affiliations & Notes
  • T. M. Curtis
    Centre for Vision Sciences, Queens Univ Belfast, Belfast, United Kingdom
  • J. McKee
    Centre for Vision Sciences, Queens Univ Belfast, Belfast, United Kingdom
  • D. P. Dash
    Centre for Vision Sciences, Queens Univ Belfast, Belfast, United Kingdom
  • A. Arora
    Centre for Vision Sciences, Queens Univ Belfast, Belfast, United Kingdom
  • C. N. Scholfield
    Centre for Vision Sciences, Queens Univ Belfast, Belfast, United Kingdom
  • J. G. McGeown
    Centre for Vision Sciences, Queens Univ Belfast, Belfast, United Kingdom
  • D. A. Simpson
    Centre for Vision Sciences, Queens Univ Belfast, Belfast, United Kingdom
  • Footnotes
    Commercial Relationships T.M. Curtis, None; J. McKee, None; D.P. Dash, None; A. Arora, None; C.N. Scholfield, None; J.G. McGeown, None; D.A. Simpson, None.
  • Footnotes
    Support Juvenile Diabetes Research Foundation, US; Wellcome Trust, UK; Fight for Sight, UK
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 2264. doi:
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      T. M. Curtis, J. McKee, D. P. Dash, A. Arora, C. N. Scholfield, J. G. McGeown, D. A. Simpson; Expression of Transient Receptor Potential Channels in Retinal Arterioles. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2264.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose:: Mammalian transient receptor potential (TRP) channels form a novel cation channel family consisting of nearly 30 members. Recent studies have suggested that these channels may play a pivotal role in controlling vascular contractility and permeability. In the present study we sought to identify systematically the TRP channel subtypes expressed in rat retinal arterioles.

Methods:: Adult male Sprague-Dawley rats were anaesthetized with CO2 and killed by cervical dislocation. Retinas were removed and intact arteriole segments isolated. TRP channel mRNA expression was evaluated by RT-PCR, and rat brain and kidney samples were used as positive controls. Immunofluorescence staining with commercially available polyclonal antibodies was also used to test for cell-specific expression of TRP channels in retinal arterioles embedded within retinal flatmount preparations. Retinas were counter-stained with propidium iodide nuclear stain. The specificity of the antibodies was investigated by parallel control experiments in the absence of primary antibody.

Results:: With RT-PCR, mRNA of 13 TRP channel subtypes was detected in retinal arterioles: TRPC1, TRPC3, TRPC4, TRPC7, TRPV1, TRPV2, TRPV4, TRPM1, TRPM2, TRPM3, TRPM7, TRPML1 and TRPML3. Lack of expression of other TRP channels could not be attributed to RT-PCR conditions since all of the other TRP channel family members could be detected in brain or kidney. Immunofluorescence labelling revealed a punctuate distribution of TRPV2 throughout the vascular smooth muscle cell layer of retinal arterioles, while TRPV4 was specifically localised to the plasma membrane of endothelial cells. Consistent with the RT-PCR results, TRPC5 protein could not be detected in retinal arterioles. This was not due to a lack of antibody reactivity because intense staining was observed in the ganglion cell layer of the retina.

Conclusions:: Our results indicate that multiple TRP channels are expressed in retinal arterioles. These data may serve as molecular basis for future work exploring the physiological significance of non-selective cation channels in the retinal vasculature.

Keywords: vascular cells • ion channels • calcium 
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