Abstract
Canonical transient receptor potential (TRPC) channels were identified as key players in maladaptive remodeling, with nuclear factor of activated T-cells (NFAT) transcription factors serving as downstream targets of TRPC-triggered Ca2+ entry in these pathological processes. Strikingly, the reconstitution of TRPC-NFAT signaling by heterologous expression yielded controversial results. Specifically, nuclear translocation of NFAT1 was found barely responsive to recombinant TRPC3, presumably based on the requirement of certain spatiotemporal signaling features. Here, we report efficient control of NFAT1 nuclear translocation in human embryonic kidney 293 (HEK293) cells by light, using a new photochromic TRPC benzimidazole activator (OptoBI-1) and a TRPC3 mutant with modified activator sensitivity. NFAT1 nuclear translocation was measured along with an all-optical protocol to record local and global Ca2+ pattern generated during light-mediated activation/deactivation cycling of TRPC3. Our results unveil the ability of wild-type TRPC3 to produce constitutive NFAT nuclear translocation. Moreover, we demonstrate that TRPC3 mutant that lacks basal activity enables spatiotemporally precise control over NFAT1 activity by photopharmacology. Our results suggest tight linkage between TRPC3 activity and NFAT1 nuclear translocation based on global cellular Ca2+ signals.
Highlights
The pathological scenario of maladaptive structural remodeling is a pivotal process in the development of severe organ pathologies, in the cardiovascular system [1,2,3]
In the first step we explored suitability of the G652A gain of function mutation for tuning of photocycling-induced TRPC3 activity pattern in HEK 293 cells
We set out to compare the temporal pattern of cation conductance generated by OptoBI-1 (10 μM)-mediated activation/deactivation cycling for TRPC3 wild-type and mutant channels
Summary
The pathological scenario of maladaptive structural remodeling is a pivotal process in the development of severe organ pathologies, in the cardiovascular system [1,2,3]. Alterations in cellular Ca2+ homeostasis, prominently featuring enhanced Ca2+ entry based on G protein-coupled receptor signaling and activation of canonical transient receptor (TRPC) channels, have been recognized as a basis for rewiring transcriptional control in response to chronic stress [2,4,5]. TRPC3/6/7 pore complexes are able to sense various stress signals, and are primarily regulated by phospholipase C (PLC)-derived lipid mediators [6,7,8], and TRPC3 stands out for its distinct constitutive activity, which is reportedly governed by its glycosylation state [9].
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