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NIH grant 1R03NS152744-01: Voltage clamp fluorometry of TMEM16A channels in axolotl oocytes
Institute: {'code': 'NS', 'abbreviation': 'NINDS', 'name': 'National Institute of Neurological Disorders and Stroke', 'admin_org_id': '12500053', 'admin_funding_url': ''}. PI: Christian Peters. Award: $155262.
Start: 2026-09-05T00:00:00.
Abstract. TMEM16A is calcium-activated chloride channel that is expressed in smooth muscle cells and
pericytes lining the brain’s blood vessels, where it contributes to the process of vasoconstriction. Hypertension
upregulates TMEM16A expression, and they become activated during ischemic stroke, exacerbating the loss of
blood flow and resulting tissue damage. As such, they are a tempting and accessible target for therapeutics to
prevent or mitigate the effects of a stroke. However, development of drugs has been hampered by the functional
complexity of TMEM16A, which has caused drugs with predicted therapeutic promise to be contraindicated
owing to unexpected pharmacology at the channel. Improved mechanistic understanding of TMEM16A
biophysics and its response to inhibitory compounds, using functional approaches informed by known 3D cryo-
EM structures of TMEM16A, would benefit future drug design and validation of existing candidates. One powerful
such approach is voltage clamp fluorometry, which combines simultaneous fluorescent tracking of
conformational changes within a protein with recording of resulting currents in order to associate structural
changes with functional outputs. However, this approach typically employs oocytes from Xenopus laevis frogs
as an expression system; the high native expression of TMEM16A in that cell type makes their use for this
approach impractical. Here, we propose to develop and test a voltage clamp fluorometry assay in oocytes from
Ambystoma mexicanum (axolotls), and to use it to characterize the structural correlates of function of TMEM16A
channels in response to salient stimuli. The overall objective of this research is to harness voltage clamp
fluorometry as both a valuable experimental approach to better understand structure-function relationships of
the TMEM16 family, and to demonstrate its value as a tool for mechanistic evaluation of the properties of
inhibitory drugs under therapeutic intervention for stroke.document fetched 2026-09-13 02:46