Pregnenolone sulphate-and cholesterol-regulated TRPM3 channels coupled to vascular smooth muscle secretion and contraction

Jacqueline Naylor, Jing Li, Carol J. Milligan, Fanning Zeng, Piruthivi Sukumar, Bing Hou, Alicia Sedo, Nadira Yuldasheva, Yasser Majeed, Dhananjay Beri, Shan Jiang, Victoria A.L. Seymour, Lynn McKeown, Bhaskar Kumar, Christian Harteneck, David O'Regan, Stephen B. Wheatcroft, Mark T. Kearney, Clare Jones, Karen E. PorterDavid J. Beech

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

RATIONALE: Transient receptor potential melastatin (TRPM)3 is a calcium-permeable ion channel activated by the neurosteroid pregnenolone sulfate and positively coupled to insulin secretion in β cells. Although vascular TRPM3 mRNA has been reported, there is no knowledge of TRPM3 protein or its regulation and function in the cardiovascular system. OBJECTIVE: To determine the relevance and regulation of TRPM3 in vascular biology. METHODS AND RESULTS: TRPM3 expression was detected at mRNA and protein levels in contractile and proliferating vascular smooth muscle cells. Calcium entry evoked by pregnenolone sulfate or sphingosine was suppressed by TRPM3 blocking antibody or knock-down of TRPM3 by RNA interference. Low-level constitutive TRPM3 activity was also detected. In proliferating cells, channel activity was coupled negatively to interleukin-6 secretion via a calcium-dependent mechanism. In freshly isolated aorta, TRPM3 positively modulated contractile responses independently of L-type calcium channels. Concentrations of pregnenolone sulfate required to evoke responses were higher than the known plasma concentrations of the steroids, leading to a screen for other stimulators. β-Cyclodextrin was one of few stimulators of TRPM3, revealing the channels to be partially suppressed by endogenous cholesterol, the precursor of pregnenolone. Elevation of cholesterol further suppressed channel activity and loading with cholesterol to generate foam cells precluded observation of TRPM3 activity. CONCLUSIONS: The data suggest functional relevance of TRPM3 in contractile and proliferating phenotypes of vascular smooth muscle cells, significance of constitutive channel activity, regulation by cholesterol, and potential value of pregnenolone sulfate in therapeutic vascular modulation.

Original languageEnglish
Pages (from-to)1507-1515
Number of pages9
JournalCirculation Research
Volume106
Issue number9
DOIs
Publication statusPublished - 14 May 2010
Externally publishedYes

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Muscle Contraction
Vascular Smooth Muscle
Cholesterol
Blood Vessels
Smooth Muscle Myocytes
Calcium
Pregnenolone
L-Type Calcium Channels
Foam Cells
Messenger RNA
Sphingosine
Blocking Antibodies
Cyclodextrins
Calcium Channels
Cardiovascular System
RNA Interference
Neurotransmitter Agents
Aorta
Interleukin-6
Proteins

Keywords

  • Calcium channel
  • Cholesterol
  • Interleukin
  • Neurosteroid
  • Transient receptor potential
  • Vascular smooth muscle

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Pregnenolone sulphate-and cholesterol-regulated TRPM3 channels coupled to vascular smooth muscle secretion and contraction. / Naylor, Jacqueline; Li, Jing; Milligan, Carol J.; Zeng, Fanning; Sukumar, Piruthivi; Hou, Bing; Sedo, Alicia; Yuldasheva, Nadira; Majeed, Yasser; Beri, Dhananjay; Jiang, Shan; Seymour, Victoria A.L.; McKeown, Lynn; Kumar, Bhaskar; Harteneck, Christian; O'Regan, David; Wheatcroft, Stephen B.; Kearney, Mark T.; Jones, Clare; Porter, Karen E.; Beech, David J.

In: Circulation Research, Vol. 106, No. 9, 14.05.2010, p. 1507-1515.

Research output: Contribution to journalArticle

Naylor, J, Li, J, Milligan, CJ, Zeng, F, Sukumar, P, Hou, B, Sedo, A, Yuldasheva, N, Majeed, Y, Beri, D, Jiang, S, Seymour, VAL, McKeown, L, Kumar, B, Harteneck, C, O'Regan, D, Wheatcroft, SB, Kearney, MT, Jones, C, Porter, KE & Beech, DJ 2010, 'Pregnenolone sulphate-and cholesterol-regulated TRPM3 channels coupled to vascular smooth muscle secretion and contraction', Circulation Research, vol. 106, no. 9, pp. 1507-1515. https://doi.org/10.1161/CIRCRESAHA.110.219329
Naylor, Jacqueline ; Li, Jing ; Milligan, Carol J. ; Zeng, Fanning ; Sukumar, Piruthivi ; Hou, Bing ; Sedo, Alicia ; Yuldasheva, Nadira ; Majeed, Yasser ; Beri, Dhananjay ; Jiang, Shan ; Seymour, Victoria A.L. ; McKeown, Lynn ; Kumar, Bhaskar ; Harteneck, Christian ; O'Regan, David ; Wheatcroft, Stephen B. ; Kearney, Mark T. ; Jones, Clare ; Porter, Karen E. ; Beech, David J. / Pregnenolone sulphate-and cholesterol-regulated TRPM3 channels coupled to vascular smooth muscle secretion and contraction. In: Circulation Research. 2010 ; Vol. 106, No. 9. pp. 1507-1515.
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AU - Naylor, Jacqueline

AU - Li, Jing

AU - Milligan, Carol J.

AU - Zeng, Fanning

AU - Sukumar, Piruthivi

AU - Hou, Bing

AU - Sedo, Alicia

AU - Yuldasheva, Nadira

AU - Majeed, Yasser

AU - Beri, Dhananjay

AU - Jiang, Shan

AU - Seymour, Victoria A.L.

AU - McKeown, Lynn

AU - Kumar, Bhaskar

AU - Harteneck, Christian

AU - O'Regan, David

AU - Wheatcroft, Stephen B.

AU - Kearney, Mark T.

AU - Jones, Clare

AU - Porter, Karen E.

AU - Beech, David J.

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N2 - RATIONALE: Transient receptor potential melastatin (TRPM)3 is a calcium-permeable ion channel activated by the neurosteroid pregnenolone sulfate and positively coupled to insulin secretion in β cells. Although vascular TRPM3 mRNA has been reported, there is no knowledge of TRPM3 protein or its regulation and function in the cardiovascular system. OBJECTIVE: To determine the relevance and regulation of TRPM3 in vascular biology. METHODS AND RESULTS: TRPM3 expression was detected at mRNA and protein levels in contractile and proliferating vascular smooth muscle cells. Calcium entry evoked by pregnenolone sulfate or sphingosine was suppressed by TRPM3 blocking antibody or knock-down of TRPM3 by RNA interference. Low-level constitutive TRPM3 activity was also detected. In proliferating cells, channel activity was coupled negatively to interleukin-6 secretion via a calcium-dependent mechanism. In freshly isolated aorta, TRPM3 positively modulated contractile responses independently of L-type calcium channels. Concentrations of pregnenolone sulfate required to evoke responses were higher than the known plasma concentrations of the steroids, leading to a screen for other stimulators. β-Cyclodextrin was one of few stimulators of TRPM3, revealing the channels to be partially suppressed by endogenous cholesterol, the precursor of pregnenolone. Elevation of cholesterol further suppressed channel activity and loading with cholesterol to generate foam cells precluded observation of TRPM3 activity. CONCLUSIONS: The data suggest functional relevance of TRPM3 in contractile and proliferating phenotypes of vascular smooth muscle cells, significance of constitutive channel activity, regulation by cholesterol, and potential value of pregnenolone sulfate in therapeutic vascular modulation.

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KW - Calcium channel

KW - Cholesterol

KW - Interleukin

KW - Neurosteroid

KW - Transient receptor potential

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