Dose- and time-dependent effects of actomyosin inhibition on live mouse outflow resistance and aqueous drainage tissues

Scientific Reports
Minhee K KoJames C H Tan

Abstract

Actomyosin contractility modulates outflow resistance of the aqueous drainage tissues and intraocular pressure, a key pathogenic factor of glaucoma. We established methodology to reliably analyze the effect of latrunculin-B (Lat-B)-induced actin depolymerization on outflow physiology in live mice. A voltage-controlled microperfusion system for delivering drugs and simultaneously analyzing outflow resistance was tested in live C57BL/6 mice. Flow rate and perfusion pressure were reproducible within a coefficient of variation of 2%. Outflow facility for phosphate-buffered saline (0.0027 ± 0.00036 μL/min/mmHg; mean ± SD) and 0.02% ethanol perfusions (Lat-B vehicle; 0.0023 ± 0.0005 μL/min/mmHg) were similar and stable over 2 hours (p > 0.1 for change), indicating absence of a 'washout' artifact seen in larger mammals. Outflow resistance changed in graded fashion, decreasing dose- and time-dependently over 2 hours for Lat-B doses of 2.5 μM (p = 0.29), 5 μM (p = 0.039) and 10 μM (p = 0.001). Resulting outflow resistance was about 10 times lower with 10 μM Lat-B than vehicle control. The filamentous actin network was decreased and structurally altered in the ciliary muscle (46 ± 5.6%) and trabecular meshwork (37 ± 8.3%) of treated eyes...Continue Reading

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Citations

Jun 9, 2015·Experimental Eye Research·Darryl R Overby, Abbot F Clark
May 18, 2016·Experimental Eye Research·Jose M GonzalezJames C H Tan
Dec 29, 2016·Progress in Retinal and Eye Research·Teresia CarreonSanjoy K Bhattacharya
Dec 8, 2017·Scientific Reports·Jose M GonzalezJames C H Tan
May 31, 2018·Investigative Ophthalmology & Visual Science·Kayla R FicarrottaChristopher L Passaglia
May 31, 2018·Investigative Ophthalmology & Visual Science·Susannah WaxmanNils A Loewen
Nov 17, 2017·Biomedicine Hub·Aleksandr YelenskiyJames C Tan
Oct 18, 2019·Survey of Ophthalmology·Ciro CostagliolaLeonardo Mastropasqua

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Methods Mentioned

BETA
fluorescence imaging
confocal microscopy
light microscopy
dissection

Software Mentioned

Excel
StatPlusMac
Minitab
NIH Image J

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