Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells.
Adequate blood flow through placental chorionic plate resistance arteries (CPAs) is necessary for oxygen and nutrient transfer to the fetus and a successful pregnancy. In non-placental vascular smooth muscle cells (SMCs), K(+) channels regulate contraction, vascular tone and blood flow. Previous stu...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3578819?pdf=render |
| _version_ | 1818179445521907712 |
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| author | Melissa F Brereton Mark Wareing Rebecca L Jones Susan L Greenwood |
| author_facet | Melissa F Brereton Mark Wareing Rebecca L Jones Susan L Greenwood |
| author_sort | Melissa F Brereton |
| collection | DOAJ |
| description | Adequate blood flow through placental chorionic plate resistance arteries (CPAs) is necessary for oxygen and nutrient transfer to the fetus and a successful pregnancy. In non-placental vascular smooth muscle cells (SMCs), K(+) channels regulate contraction, vascular tone and blood flow. Previous studies showed that K(+) channel modulators alter CPA tone, but did not distinguish between effects on K(+) channels in endothelial cells and SMCs. In this study, we developed a preparation of freshly isolated CPASMCs of normal pregnancy and investigated K(+) channel expression and function. CPASMCs were isolated from normal human term placentas using enzymatic digestion. Purity and phenotype was confirmed with immunocytochemistry. Whole-cell patch clamp was used to assess K(+) channel currents, and mRNA and protein expression was determined in intact CPAs and isolated SMCs with RT-PCR and immunostaining. Isolated SMCs expressed α-actin but not CD31, a marker of endothelial cells. CPASMCs and intact CPAs expressed h-caldesmon and non-muscle myosin heavy chain-2; phenotypic markers of contractile and synthetic SMCs respectively. Whole-cell currents were inhibited by 4-AP, TEA, charybdotoxin and iberiotoxin implicating functional K(v) and BK(Ca) channels. 1-EBIO enhanced whole cell currents which were abolished by TRAM-34 and reduced by apamin indicating activation of IK(Ca) and SK(Ca) respectively. BK(Ca), IK(Ca) and SK(Ca)3 mRNA and/or protein were expressed in CPASMCs and intact CPAs. This study provides the first direct evidence for functional K(v), BK(Ca,) IK(Ca) and SK(Ca) channels in CPASMCs. These cells display a mixed phenotype implicating a dual role for CPASMCs in controlling both fetoplacental vascular resistance and vasculogenesis. |
| first_indexed | 2024-12-11T21:03:59Z |
| format | Article |
| id | doaj.art-343940e8ba5d4b76bddbae3052db540c |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T21:03:59Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-343940e8ba5d4b76bddbae3052db540c2022-12-22T00:50:55ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0182e5745110.1371/journal.pone.0057451Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells.Melissa F BreretonMark WareingRebecca L JonesSusan L GreenwoodAdequate blood flow through placental chorionic plate resistance arteries (CPAs) is necessary for oxygen and nutrient transfer to the fetus and a successful pregnancy. In non-placental vascular smooth muscle cells (SMCs), K(+) channels regulate contraction, vascular tone and blood flow. Previous studies showed that K(+) channel modulators alter CPA tone, but did not distinguish between effects on K(+) channels in endothelial cells and SMCs. In this study, we developed a preparation of freshly isolated CPASMCs of normal pregnancy and investigated K(+) channel expression and function. CPASMCs were isolated from normal human term placentas using enzymatic digestion. Purity and phenotype was confirmed with immunocytochemistry. Whole-cell patch clamp was used to assess K(+) channel currents, and mRNA and protein expression was determined in intact CPAs and isolated SMCs with RT-PCR and immunostaining. Isolated SMCs expressed α-actin but not CD31, a marker of endothelial cells. CPASMCs and intact CPAs expressed h-caldesmon and non-muscle myosin heavy chain-2; phenotypic markers of contractile and synthetic SMCs respectively. Whole-cell currents were inhibited by 4-AP, TEA, charybdotoxin and iberiotoxin implicating functional K(v) and BK(Ca) channels. 1-EBIO enhanced whole cell currents which were abolished by TRAM-34 and reduced by apamin indicating activation of IK(Ca) and SK(Ca) respectively. BK(Ca), IK(Ca) and SK(Ca)3 mRNA and/or protein were expressed in CPASMCs and intact CPAs. This study provides the first direct evidence for functional K(v), BK(Ca,) IK(Ca) and SK(Ca) channels in CPASMCs. These cells display a mixed phenotype implicating a dual role for CPASMCs in controlling both fetoplacental vascular resistance and vasculogenesis.http://europepmc.org/articles/PMC3578819?pdf=render |
| spellingShingle | Melissa F Brereton Mark Wareing Rebecca L Jones Susan L Greenwood Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells. PLoS ONE |
| title | Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells. |
| title_full | Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells. |
| title_fullStr | Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells. |
| title_full_unstemmed | Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells. |
| title_short | Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells. |
| title_sort | characterisation of k channels in human fetoplacental vascular smooth muscle cells |
| url | http://europepmc.org/articles/PMC3578819?pdf=render |
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