Reduced RhoA activity mediates acute alcohol intoxication-induced inhibition of lymphatic myogenic constriction despite increased cytosolic [Ca(2+) ].

OBJECTIVES: We previously showed that AAI reduces lymphatic myogenic constriction in response to step increases in luminal pressure. Because of the known role of Ca(2+) in smooth muscle contractile responses, we investigated how alcohol impacts cyclic Ca(2+) and whether changes in RhoA/ROCK-mediated Ca(2+) sensitivity underlie the alcohol-induced reduction of myogenic responsiveness.
METHODS: AAI was produced by intragastric administration of 30% alcohol in rats. Mesenteric lymphatics were cannulated and loaded with Fura-2 AM to [Ca(2+) ]i for 30 minutes after AAI. Active GTP-bound RhoA levels were determined by ELISA. To determine ROCK's ability to restore myogenic responsiveness following AAI, isolated lymphatics were transfected with constitutively active ca-ROCK protein.
RESULTS: Lymphatics from alcohol-treated rats displayed significantly larger Ca(2+) transients. Also, step increases in luminal pressure caused a gradual rise in the basal [Ca(2+) ]i between transients that was greater in lymphatics submitted to AAI, compared to vehicle control. RhoA-GTP was significantly reduced in lymphatics from the AAI group, compared to vehicle control. Transfection with ca-ROCK protein restored the myogenic response of lymphatic vessels isolated from AAI animals.
CONCLUSIONS: The data strongly suggest that the alcohol-induced inhibition of mesenteric lymphatic myogenic constriction is mediated by reduced RhoA/ROCK-mediated Ca(2+) sensitivity.