Nax-positive glial cells in the organum vasculosum laminae terminalis produce epoxyeicosatrienoic acids to induce water intake in response to increases in [Na+] in body fluids

Abstract Na x is a [Na + ] sensor expressed in specific glial cells in the sensory circumventricular organs (sCVOs) in the brain. We recently demonstrated that Na x signals are involved in the control of not only salt intake but also water intake behavior. Our pharmacological experiments suggested that Na x signals led to activation of neurons bearing TRPV4 by using epoxyeicosatrienoic acids (EETs) as gliotransmitters to stimulate water intake. In the present study, we performed selective lesions of individual sCVOs in wild-type (WT) mice and the site-directed rescue of Na x expression in Na x -gene knockout ( Na x -KO) mice. These experiments revealed that the Na x channel in the organum vasculosum laminae terminalis (OVLT) functions as a [Na + ] sensor for the control of water intake behavior. Direct measurements of 5,6-EET and 8,9-EET in the OVLT demonstrated that EET levels were indeed increased two-fold by water deprivation for two days in WT, but not Na x -KO mice, indicating that EETs were Na x -dependently produced in the OVLT in response to increases in [Na + ] in body fluids. More importantly, intracerebroventricular injection of 5,6-EET at the same level was effective to induce water intake. Double staining revealed that Na x -positive cells also expressed Cyp2c44, a cytochrome P450 epoxygenase, to generate EETs. Collectively, these results indicate that Na x -positive glial cells produce EETs to activate TRPV4-positive neurons which may stimulate water intake, in response to increases in [Na + ] of body fluids.