May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Effect of Adenosine on P2X7 Receptor-mediated Ca2+ Signalling in Human Muller Cells and Retinal Neurons
Author Affiliations & Notes
  • J. Sanderson
    School of Biological Science, University of East Anglia, Norwich, United Kingdom
  • D.J. Collison
    School of Biological Science, University of East Anglia, Norwich, United Kingdom
  • J.A. Eldred
    School of Biological Science, University of East Anglia, Norwich, United Kingdom
  • G. Duncan
    School of Biological Science, University of East Anglia, Norwich, United Kingdom
  • D.C. Broadway
    Department of Ophthalmology, Norfolk and Norwich University Hospital, Norwich, United Kingdom
  • Footnotes
    Commercial Relationships  J. Sanderson, None; D.J. Collison, None; J.A. Eldred, None; G. Duncan, None; D.C. Broadway, None.
  • Footnotes
    Support  RNIB, Glaucoma Research, Fight for Sight
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 5204. doi:
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      J. Sanderson, D.J. Collison, J.A. Eldred, G. Duncan, D.C. Broadway; Effect of Adenosine on P2X7 Receptor-mediated Ca2+ Signalling in Human Muller Cells and Retinal Neurons . Invest. Ophthalmol. Vis. Sci. 2003;44(13):5204.

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

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Abstract

Abstract: : Purpose: The P2X7 ATP receptor is proposed to play a role in apoptosis. Functional expression of this receptor in human retinal cells in culture was investigated using the P2X7 agonist BzATP. Adenosine is a proposed neuromodulator in the retina. The effect of adenosine on BzATP-induced Ca2+ signalling was also investigated. Methods: Human retinas were obtained from donor eyes within 24 hours post mortem. The neural retina was dissociated using papain, plated onto laminin-coated cover slips and cultured in Ham/F12 medium supplemented with 10% FCS and 10ng/ml bFGF. Ca2+ measurements were made using ratiometric digital imaging techniques with the Ca2+-indicator Fura-2. Results: BzATP (50µ M) induced an increase in intracellular Ca2+ in Müller cells and in a subset of retinal neurons. This response was totally inhibited by the P2X antagonist PPADS (100µ M), while the L-type Ca2+ channel antagonist nifedipine (10µ M) had little effect on the BzATP-induced Ca2+ signal. Application of adenosine (10µ M) had no effect on resting intracellular Ca2+ concentration in the BzATP-sensitive cells. However, in the presence of adenosine the Ca2+ response to BzATP was potentiated in both Müller and neuronal cells. Moreover, it was observed that neurons responding only minimally to BzATP, gave large Ca2+ responses in the presence of both BzATP and adenosine. The adenosine-induced potentiation was inhibited by the A1 receptor antagonist DPCPX (1µ M), whereas the A3 receptor antagonist MRS1191 (1µ M) had no effect. Conclusions: P2X7 ATP receptors are functionally expressed in cultured human retinal Müller and neuronal cells. Adenosine potentiates the BzATP-induced Ca2+ signal via A1 adenosine receptors. This enhanced Ca2+ response may have implications for neuronal cell death, such as that occurring in glaucoma.

Keywords: retinal culture • calcium • adenosine 
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