May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
RT–PCR Mapping of Serotonin Receptor Subtype mRNAs in Human Ciliary Body and Trabecular Meshwork
Author Affiliations & Notes
  • N.A. Sharif
    Molecular Pharmacology R2–19, Alcon Research Ltd, Fort Worth, TX
  • C.R. Kelly
    Molecular Pharmacology R2–19, Alcon Research Ltd, Fort Worth, TX
  • J.Y. Crider
    Molecular Pharmacology R2–19, Alcon Research Ltd, Fort Worth, TX
  • M. Senchyna
    University of Waterloo, Waterloo, ON, Canada
  • Footnotes
    Commercial Relationships  N.A. Sharif, Alcon Research, Ltd E; Alcon Research, Ltd E; Alcon Research, Ltd E; Alcon Research, Ltd. C; C.R. Kelly, Alcon Research, Ltd. E; J.Y. Crider, Alcon Research, Ltd. E; M. Senchyna, Alcon Research, Ltd C.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 3688. doi:
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      N.A. Sharif, C.R. Kelly, J.Y. Crider, M. Senchyna; RT–PCR Mapping of Serotonin Receptor Subtype mRNAs in Human Ciliary Body and Trabecular Meshwork . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3688.

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

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Abstract
 
Abstract:
 

To study the relative distribution of the mRNAs for serotonin (5HT) receptor subtypes in human ocular tissues / cells using reverse transcriptase polymerase chain reaction (RT–PCR) and determine receptor–mediated functional responses in isolated human ocular cells. Standard RT–PCR procedures were utilized for the mRNA detection (up to 10 human donor eyes for tissues; n = 8 donor human trabecular meshwork (h–TM) cell samples). Phosphoinositide (PI) hydrolysis and intracellular Ca2+ ([Ca2+]i ) mobilization were determined using previously published methods.

 

 

5HT2 receptor–mediated PI turnover [(R)–DOI EC50 = 13 ± 5 nM; 5HT EC50 = 364 ± 75 nM)] and [Ca2+]i mobilization [(R)–DOI EC50 = 18 ± 6 nM; 5HT EC50 = 40 ± 5 nM)] was found in h–TM cells. These responses were antagonized by a 5HT2Areceptor antagonist (M–100907; Ki = 39 pM against PI and 1 nM against [Ca2+]i mobilization). Similar data were obtained for the human ciliary muscle cells isolated from several donor eyes (5HT EC50 = 95 ±15 nM in PI assays; EC50 = 215 nM in [Ca2+]i mobilization assays). The most abundant receptor mRNAs were for the 5HT2A/B and 5HT7 receptors in human ciliary body/ ciliary muscle (CM), iris and in h–TM cells. The presence of functional 5HT2 receptors in h–TM and h–CM cells can help explain the potent IOP–lowering activity observed with 5HT2 agonists such as R–DOI (34% at 300 µg) and α–methyl–5HT (33% at 300 µg) following topical ocular dosing in concious ocular hypertensive cynomolgus monkeys. CR : E

 

 
Keywords: receptors: pharmacology/physiology • receptors • signal transduction: pharmacology/physiology 
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