WNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons

The activation of chloride (Cl−)permeable gamma (γ)-aminobutyric acid type A(GABAA) receptors induces synaptic inhibition in mature and excitation in immature neurons. This developmental “switch” in GABA function controlled by its polarity depends on the postnatal decrease in intraneuronal Cl− conce...

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Autores principales:	Wee Meng Lim, Eunice W. M. Chin, Bor Luen Tang, Tingting Chen, Eyleen L. K. Goh
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	WNK3 GABAergic inhibitory tone synaptic excitation hyperpolarized EGABA neuronal excitability KCC2 cotransporter Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571
Acceso en línea:	https://doaj.org/article/0dd72a69fbdb4a22be61103f73e5164f
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spelling	oai:doaj.org-article:0dd72a69fbdb4a22be61103f73e5164f2021-11-10T07:12:56ZWNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons1662-509910.3389/fnmol.2021.762142https://doaj.org/article/0dd72a69fbdb4a22be61103f73e5164f2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fnmol.2021.762142/fullhttps://doaj.org/toc/1662-5099The activation of chloride (Cl−)permeable gamma (γ)-aminobutyric acid type A(GABAA) receptors induces synaptic inhibition in mature and excitation in immature neurons. This developmental “switch” in GABA function controlled by its polarity depends on the postnatal decrease in intraneuronal Cl− concentration mediated by KCC2, a member of cation-chloride cotransporters (CCCs). The serine-threonine kinase WNK3 (With No Lysine [K]), is a potent regulator of all CCCs and is expressed in neurons. Here, we characterized the functions of WNK3 and its role in GABAergic signaling in cultured embryonic day 18 (E18) hippocampal neurons. We observed a decrease in WNK3 expression as neurons mature. Knocking down of WNK3 significantly hyperpolarized EGABA in mature neurons (DIV13–15) but had no effect on immature neurons (DIV6–8). This hyperpolarized EGABA in WNK3-deficient neurons was not due to the total expression of NKCC1 and KCC2, that remained unchanged. However, there was a reduction in phosphorylated KCC2 at the membrane, suggesting an increase in KCC2 chloride export activity. Furthermore, hyperpolarized EGABA observed in WNK3-deficient neurons can be reversed by the KCC2 inhibitor, VU024055, thus indicating that WNK3 acts through KCC2 to influence EGABA. Notably, WNK3 knockdown resulted in morphological changes in mature but not immature neurons. Electrophysiological characterization of WNK3-deficient mature neurons revealed reduced capacitances but increased intrinsic excitability and synaptic excitation. Hence, our study demonstrates that WNK3 maintains the “adult” GABAergic inhibitory tone in neurons and plays a role in the morphological development of neurons and excitability.Wee Meng LimWee Meng LimEunice W. M. ChinEunice W. M. ChinEunice W. M. ChinBor Luen TangBor Luen TangTingting ChenEyleen L. K. GohEyleen L. K. GohFrontiers Media S.A.articleWNK3GABAergic inhibitory tonesynaptic excitationhyperpolarized EGABAneuronal excitabilityKCC2 cotransporterNeurosciences. Biological psychiatry. NeuropsychiatryRC321-571ENFrontiers in Molecular Neuroscience, Vol 14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	WNK3 GABAergic inhibitory tone synaptic excitation hyperpolarized EGABA neuronal excitability KCC2 cotransporter Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571
spellingShingle	WNK3 GABAergic inhibitory tone synaptic excitation hyperpolarized EGABA neuronal excitability KCC2 cotransporter Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 Wee Meng Lim Wee Meng Lim Eunice W. M. Chin Eunice W. M. Chin Eunice W. M. Chin Bor Luen Tang Bor Luen Tang Tingting Chen Eyleen L. K. Goh Eyleen L. K. Goh WNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons
description	The activation of chloride (Cl−)permeable gamma (γ)-aminobutyric acid type A(GABAA) receptors induces synaptic inhibition in mature and excitation in immature neurons. This developmental “switch” in GABA function controlled by its polarity depends on the postnatal decrease in intraneuronal Cl− concentration mediated by KCC2, a member of cation-chloride cotransporters (CCCs). The serine-threonine kinase WNK3 (With No Lysine [K]), is a potent regulator of all CCCs and is expressed in neurons. Here, we characterized the functions of WNK3 and its role in GABAergic signaling in cultured embryonic day 18 (E18) hippocampal neurons. We observed a decrease in WNK3 expression as neurons mature. Knocking down of WNK3 significantly hyperpolarized EGABA in mature neurons (DIV13–15) but had no effect on immature neurons (DIV6–8). This hyperpolarized EGABA in WNK3-deficient neurons was not due to the total expression of NKCC1 and KCC2, that remained unchanged. However, there was a reduction in phosphorylated KCC2 at the membrane, suggesting an increase in KCC2 chloride export activity. Furthermore, hyperpolarized EGABA observed in WNK3-deficient neurons can be reversed by the KCC2 inhibitor, VU024055, thus indicating that WNK3 acts through KCC2 to influence EGABA. Notably, WNK3 knockdown resulted in morphological changes in mature but not immature neurons. Electrophysiological characterization of WNK3-deficient mature neurons revealed reduced capacitances but increased intrinsic excitability and synaptic excitation. Hence, our study demonstrates that WNK3 maintains the “adult” GABAergic inhibitory tone in neurons and plays a role in the morphological development of neurons and excitability.
format	article
author	Wee Meng Lim Wee Meng Lim Eunice W. M. Chin Eunice W. M. Chin Eunice W. M. Chin Bor Luen Tang Bor Luen Tang Tingting Chen Eyleen L. K. Goh Eyleen L. K. Goh
author_facet	Wee Meng Lim Wee Meng Lim Eunice W. M. Chin Eunice W. M. Chin Eunice W. M. Chin Bor Luen Tang Bor Luen Tang Tingting Chen Eyleen L. K. Goh Eyleen L. K. Goh
author_sort	Wee Meng Lim
title	WNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons
title_short	WNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons
title_full	WNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons
title_fullStr	WNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons
title_full_unstemmed	WNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons
title_sort	wnk3 maintains the gabaergic inhibitory tone, synaptic excitation and neuronal excitability via regulation of kcc2 cotransporter in mature neurons
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/0dd72a69fbdb4a22be61103f73e5164f
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WNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons

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