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Robert J. Wordinger, Wendi Lambert, Rajnee Agarwal, Mihir Talati, Abbot F. Clark; Human Trabecular Meshwork Cells Secrete Neurotrophins and Express Neurotrophin Receptors (Trk). Invest. Ophthalmol. Vis. Sci. 2000;41(12):3833-3841.
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purpose. The purpose of this study was to compare the mRNA expression of
neurotrophins (NTs) and NT receptors (Trk) in cultured human trabecular
meshwork (HTM) cells and ex vivo HTM tissues, to immunolocalize both NT
and Trk receptors in cultured HTM cells, and to demonstrate secretion
of NTs by HTM cells.
methods. Reverse transcription–polymerase chain reaction (RT-PCR) was used to
detect the expression of NT and Trk receptor mRNAs in early-passaged,
cultured HTM cells from donors of several ages. RT-PCR was used on ex
vivo HTM tissues from donors to compare and contrast mRNA expression
with cell culture results. In addition, immunohistochemistry was used
to localize the translated NT and low- (p75) and high- (Trk) affinity
NT receptor proteins within cultured HTM cells and trabecular meshwork
tissues. Last, enzyme-linked immunoassay (ELISA) was used to
demonstrate secretion of NTs by HTM cells.
results. Amplification products of the expected size for NTs were detected in
both cultured HTM cells and ex vivo HTM tissues. Specifically
identified were amplification products for the following NTs: NGF,
BDNF, NT-3, and NT-4. Amplification products for the full-length Trk A
and Trk C high-affinity receptor were observed, as well as truncated
isoforms for Trk B and Trk C. No amplification products were produced
for the full-length Trk B receptor nor for the low-affinity p75
receptor. Immunohistochemistry indicated that proteins for the various
NTs and full-length and truncated Trk receptors were translated by
cultured HTM cells and tissues. Immunoassays (ELISA) detected BDNF,
NT-4, NGF, and NT-3 in the culture media from HTM cells.
conclusions. The results demonstrate, for the first time, mRNA expression for NT and
Trk receptors by both cultured HTM cells and ex vivo HTM tissues. NTs
were immunolocalized in HTM tissues and cultured HTM cells are capable
of secreting NTs. Specific NTs acting through high-affinity Trk
receptors within the HTM may be involved in maintaining the normal
function of this complex tissue.
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