Investigating the role of local acidosis in hypothalamic vasopressin cell excitation during hypoxia

The supraoptic nucleus (SON) of the hypothalamus plays a key role in the regulation of fluid homeostasis through the synthesis and release of vasopressin (VP). In this population, VP release can occur at both dendritic and axonal sites, allowing for both localized and distal effects such as renal water retention and vasoconstriction. Recent work from our laboratory demonstrated an inverse neurovascular coupling (iNVC) response in which VP cell activation by acute salt loading resulted in dendritic release of VP, leading to localized vasoconstriction and hypoxia. Additionally, we demonstrated that the hypoxic milieu in turn potentiates VP cell activity and increases firing, suggesting that these mechanism constitutes a positive feedback loop that optimized VP activation in order to cope with the homeostatic challenge (Roy et al., Cell Reports 2021). Still, the biophysical mechanism by which the iNVC-hypoxia elicits increased firing in VP neurons has not yet been explored. We hypothesized that hypoxia-induced acidosis, via activation of acid sensitive ion channels (ASICs) (which are present in SON VP neurons; Ohbuchi et al in 2010) is a key mechanism contributing to VP cell excitation in the iNVC positive feedback loop. We investigated this with whole cell patch clamp electrophysiology using eGFP-VP transgenic rats. We recorded from identified SON VP cells and measured deflections in voltage as bath pH was changed from 7.25 to 6.3 (10 mins). Bath solution was either HCL-buffered artificial cerebrospinal fluid (aCSF) aerated with 95% O 2 /5% CO 2 or HEPES-buffered solution (HBS). We observed an excitatory response to the decreases in pH in most SON VP cells recorded (68.75%, n = 11/16). These responses were reversible and, in many cases, repeatable following a second stimulation. To examine cell-type specificity, ongoing studies are focusing on comparing these responses to those obtained from oxytocin cells. Further, we plan to repeat these experiments in the presence of the ASIC-blocker amiloride and determine whether the hypoxia-induced excitatory effect during iNVC is indeed dependent on ASIC-channels. Together, our results support our hypothesis that tissue acidosis (likely via activation of ASIC-channels) contributes to the excitatory feedback loop evoked during salt-induced iNVC in VP neurons. This abstract was supported by the National Heart Lung and Blood Institute of the National Institutes of Health R01 HL162575-01 (J.E.S.). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.