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Fumiko Taniguchi, Yasuyuki Suzuki, Hiroki Kurihara, Yukiko Kurihara, Hiroyoshi Kasai, Shiroaki Shirato, Makoto Araie; Molecular Cloning of the Bovine MYOC and Induction of Its Expression in Trabecular Meshwork Cells. Invest. Ophthalmol. Vis. Sci. 2000;41(8):2070-2075.
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purpose. Myocilin gene (MYOC) was identified as one of the
disease-causing genes of primary open-angle glaucoma. This study was
conducted to establish a system for the investigation of the biological
role of MYOC in vitro by using bovine eyes, which are easy
to obtain and have been widely used to examine the aqueous outflow
system. The cDNA sequence of the bovine MYOC was determined
and its expression in bovine eyes was examined with a quantitative
polymerase chain reaction (PCR) assay.
methods. Bovine MYOC cDNA was obtained from cultured bovine
trabecular meshwork cells, and part of its sequence was determined
using a primer pair designed based on the known sequence of the human MYOC gene. The 3′ and 5′ ends of this sequence were
determined using the method of 3′ and 5′ rapid amplification of cDNA
ends. The induction of the MYOC gene in cultured bovine
trabecular meshwork cells after exposure to dexamethasone was
quantitatively examined with real-time quantitative PCR using a probe
designed according to the sequence of the determined bovine MYOC gene.
results. Bovine MYOC protein was composed of 490 amino acids, which was 81.6%
identical with that of human MYOC protein. Most of the amino acid
residues of which mutation was reported to cause glaucoma were
conserved in the bovine MYOC protein. After 2 weeks of treatment with
500 nM dexamethasone, expression of bovine MYOC mRNA was
amplified 14-fold (14.1 ± 5.1-fold, mean ± SEM) measured by
real-time quantitative PCR.
conclusions. The cDNA sequence of the bovine MYOC gene had a high degree
of similarity to that of the human MYOC gene. Investigation
of the function of bovine MYOC may contribute to identifying
the role of MYOC protein in the aqueous outflow
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