Purchase this article with an account.
Phi T Ho, Genea Edwards, Fareed Rifai, Sanjoy K Bhattacharya; Changes in the levels of selected mechanosensing channels in normotensive versus hypertensive trabecular meshwork. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5720.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To determine the changes in the levels of selected mechanosensing channels (Piezo1-2, TASK1, TREK-1, TRPC1-3, TRPC6, TRPM2, TRPP2) in the trabecular meshwork (TM) of DBA/2J mice in normotensive versus hypertensive state.
All studies were performed adhering to the ARVO statement for use of animals in vision research under IACUC approved protocols. DBA/2J mice of age 3-9 months (n=40 at each point) was used for these studies. Intraocular pressure (IOP) was measured using a TonoLab (Colonial Medical Supplies, NH). DBA/2J mice were categorized into normotensive (IOP ≤15mmHg; usually in mice with less than 7.5 months of age) and hypertensive (IOP range of 18-36mmHg) groups. Quantitative Western Blot and immunohistochemistry were carried out using commercial antibodies to these channels (as noted above). These channels were previously found in the DBA/2J TM by microarray and immunohistochemical analyses in our laboratory. Non-amplified IR-coupled secondary antibody was used for Western Blots. Confocal microscopy on a Leica SP5 microscope was used for quantification of immunohistochemical signals. Quantitative label free mass spectrometry was also carried out to validate the relative quantification of these channels.
Selected mechanosensing channels (Piezo1&2, TASK1, TREK-1, TRPC1-3, TRPC6, TRPM2, TRPP2) showed at least a 30% difference in levels of expression between normotensive and hypertensive states (normalized with beta-actin) determined by Quantitative Western Blot analysis and corroborated by other methods.
Our investigation confirms the modulation of selected mechanosensing channels in TM tissue in different states of IOP.
This PDF is available to Subscribers Only