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Willem Kamphuis, Andrea Schneemann, Luc M. van Beek, August B. Smit, Philip F. J. Hoyng, Eisuke Koya; Prostanoid Receptor Gene Expression Profile in Human Trabecular Meshwork: A Quantitative Real-Time PCR Approach. Invest. Ophthalmol. Vis. Sci. 2001;42(13):3209-3215.
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purpose. To assess the expression pattern of prostanoid receptor–encoding genes
in trabecular meshwork (TM) of human donor eyes.
methods. Disposed human donor eyes (n = 10) were obtained
from the Cornea Bank, Amsterdam. The TM was dissected from the scleral
tissue and homogenized in lysis buffer, and total RNA was isolated. The
RNA was converted into cDNA and used as a template for noncompetitive
quantitative real-time polymerase chain reaction (PCR) using green
fluorescent dye to quantify the accumulation of double-stranded PCR
product. Specific primers for four housekeeping genes and DP,
EP1, EP2, EP3, EP4, FP,
IP, and TP receptor–encoding transcripts were developed and tested for
results. The characterized expression profile was highly reproducible in all
samples, with the EP2 receptor–encoding transcript in the
highest abundance, followed by FP, TP, IP, and EP4 at
levels that were approximately 10 to 15 times lower than that of the
EP2 subtype. DP and EP3 were at the lowest
levels, which were, on average, 45 times and 228 times lower than
conclusions. These data show that all prostanoid receptors are expressed at
different levels in human TM tissue. Because the gene expression of the
EP2 receptor is, on average, 15 times more abundant than
that of the EP4 receptor, it may be expected that the
increase in flow and cAMP levels in response to the activation of the
EP receptors by application of prostaglandin E1 (PGE1), is primarily mediated by the EP2 receptor. These data should be considered when designing
prostanoid receptor mimetics intended to enhance the aqueous humor
outflow through the TM and Schlemm’s canal.
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