Rho Kinase Enhances Contractions of Rat Mesenteric Collecting Lymphatics
Identifieur interne : 002568 ( Main/Exploration ); précédent : 002567; suivant : 002569Rho Kinase Enhances Contractions of Rat Mesenteric Collecting Lymphatics
Auteurs : Kristine H. Kurtz [États-Unis] ; Flavia M. Souza-Smith [États-Unis] ; Andrea N. Moor [États-Unis] ; Jerome W. Breslin [États-Unis]Source :
- PLoS ONE [ 1932-6203 ] ; 2014.
Abstract
The mechanisms that control phasic and tonic contractions of lymphatic vessels are poorly understood. We hypothesized that rho kinase ROCK, previously shown to increase calcium (Ca2+) sensitivity in vascular smooth muscle, enhances lymphatic contractile activity in a similar fashion. Contractions of isolated rat mesenteric lymphatic vessels were observed at a luminal pressure of 2 cm H2O in a 37°C bath. The expression of ROCK in isolated rat mesenteric lymphatic vessels was assessed by Western blotting and confocal microscopy. The role of ROCK in contractile function was tested using two specific yet structurally distinct inhibitors: H1152 (0.1–10 μM) and Y-27632 (0.5–50 μM). In addition, lymphatics were transfected with constitutively active (ca)-ROCK protein (2 μg/ml) to assess gain of contractile function. Vessel diameter and the concentration of intracellular free Ca2+ ([Ca2+]i) were simultaneously measured in a subset of isolated lymphatics loaded with the Ca2+-sensing dye fura-2. The results show expression of both the ROCK1 and ROCK2 isoforms in lymphatic vessels. Inhibition of ROCK increased lymphatic end diastolic diameter and end systolic diameter in a concentration-dependent manner. Significant reductions in lymphatic tone and contraction amplitude were observed after treatment 1–10 μM H1152 or 25–50 μM Y-27632. H1152 (10 μM) also significantly reduced contraction frequency. Transient increases in [Ca2+]i preceded each phasic contraction, however this pattern was disrupted by either 10 μM H1152 or 50 μM Y-27632 in the majority of lymphatics studied. The significant decrease in tone caused by H1152 or Y-27632 was not associated with a significant change in the basal [Ca2+]i between transients. Transfection with ca-ROCK protein enhanced lymphatic tone, but was not associated with a significant change in basal [Ca2+]i. Our data suggest that ROCK mediates normal tonic constriction and influences phasic contractions in lymphatics. We propose that ROCK modulates Ca2+ sensitivity of contractile proteins in lymphatics.
Url:
DOI: 10.1371/journal.pone.0094082
PubMed: 24710574
PubMed Central: 3978029
Affiliations:
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Kurtz</name><affiliation wicri:level="2"><nlm:aff id="aff1"><addr-line>Department of Physiology, School of Medicine, Louisiana State University Health Sciences Center, New Orleans, Louisiana, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Physiology, School of Medicine, Louisiana State University Health Sciences Center, New Orleans, Louisiana</wicri:regionArea><placeName><region type="state">Louisiane</region></placeName></affiliation></author><author><name sortKey="Souza Smith, Flavia M" sort="Souza Smith, Flavia M" uniqKey="Souza Smith F" first="Flavia M." last="Souza-Smith">Flavia M. Souza-Smith</name><affiliation wicri:level="2"><nlm:aff id="aff1"><addr-line>Department of Physiology, School of Medicine, Louisiana State University Health Sciences Center, New Orleans, Louisiana, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Physiology, School of Medicine, Louisiana State University Health Sciences Center, New Orleans, Louisiana</wicri:regionArea><placeName><region type="state">Louisiane</region></placeName></affiliation></author><author><name sortKey="Moor, Andrea N" sort="Moor, Andrea N" uniqKey="Moor A" first="Andrea N." last="Moor">Andrea N. Moor</name><affiliation wicri:level="4"><nlm:aff id="aff2"><addr-line>Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, Tampa, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, Tampa, Florida</wicri:regionArea><placeName><region type="state">Floride</region><settlement type="city">Tampa</settlement></placeName><orgName type="university">Université de Floride du Sud</orgName></affiliation></author><author><name sortKey="Breslin, Jerome W" sort="Breslin, Jerome W" uniqKey="Breslin J" first="Jerome W." last="Breslin">Jerome W. Breslin</name><affiliation wicri:level="4"><nlm:aff id="aff2"><addr-line>Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, Tampa, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, Tampa, Florida</wicri:regionArea><placeName><region type="state">Floride</region><settlement type="city">Tampa</settlement></placeName><orgName type="university">Université de Floride du Sud</orgName></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The mechanisms that control phasic and tonic contractions of lymphatic vessels are poorly understood. We hypothesized that rho kinase ROCK, previously shown to increase calcium (Ca<sup>2+</sup>) sensitivity in vascular smooth muscle, enhances lymphatic contractile activity in a similar fashion. Contractions of isolated rat mesenteric lymphatic vessels were observed at a luminal pressure of 2 cm H<sub>2</sub>O in a 37°C bath. The expression of ROCK in isolated rat mesenteric lymphatic vessels was assessed by Western blotting and confocal microscopy. The role of ROCK in contractile function was tested using two specific yet structurally distinct inhibitors: H1152 (0.1–10 μM) and Y-27632 (0.5–50 μM). In addition, lymphatics were transfected with constitutively active (ca)-ROCK protein (2 μg/ml) to assess gain of contractile function. Vessel diameter and the concentration of intracellular free Ca<sup>2+</sup> ([Ca<sup>2+</sup>]<sub>i</sub>) were simultaneously measured in a subset of isolated lymphatics loaded with the Ca<sup>2+</sup>-sensing dye fura-2. The results show expression of both the ROCK1 and ROCK2 isoforms in lymphatic vessels. Inhibition of ROCK increased lymphatic end diastolic diameter and end systolic diameter in a concentration-dependent manner. Significant reductions in lymphatic tone and contraction amplitude were observed after treatment 1–10 μM H1152 or 25–50 μM Y-27632. H1152 (10 μM) also significantly reduced contraction frequency. Transient increases in [Ca<sup>2+</sup>]<sub>i</sub> preceded each phasic contraction, however this pattern was disrupted by either 10 μM H1152 or 50 μM Y-27632 in the majority of lymphatics studied. The significant decrease in tone caused by H1152 or Y-27632 was not associated with a significant change in the basal [Ca<sup>2+</sup>]<sub>i</sub> between transients. Transfection with ca-ROCK protein enhanced lymphatic tone, but was not associated with a significant change in basal [Ca<sup>2+</sup>]<sub>i</sub>. Our data suggest that ROCK mediates normal tonic constriction and influences phasic contractions in lymphatics. We propose that ROCK modulates Ca<sup>2+</sup> sensitivity of contractile proteins in lymphatics.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Von Der Weid, Py" uniqKey="Von Der Weid P">PY von der Weid</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wang, Y" uniqKey="Wang Y">Y Wang</name></author><author><name sortKey="Oliver, G" uniqKey="Oliver G">G Oliver</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tammela, T" uniqKey="Tammela T">T Tammela</name></author><author><name sortKey="Alitalo, K" uniqKey="Alitalo K">K Alitalo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Imtiaz, Ms" uniqKey="Imtiaz M">MS Imtiaz</name></author><author><name sortKey="Zhao, J" uniqKey="Zhao J">J Zhao</name></author><author><name sortKey="Hosaka, K" uniqKey="Hosaka K">K Hosaka</name></author><author><name sortKey="Von Der Weid, Py" 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Kurtz</name></region><name sortKey="Breslin, Jerome W" sort="Breslin, Jerome W" uniqKey="Breslin J" first="Jerome W." last="Breslin">Jerome W. Breslin</name><name sortKey="Moor, Andrea N" sort="Moor, Andrea N" uniqKey="Moor A" first="Andrea N." last="Moor">Andrea N. Moor</name><name sortKey="Souza Smith, Flavia M" sort="Souza Smith, Flavia M" uniqKey="Souza Smith F" first="Flavia M." last="Souza-Smith">Flavia M. Souza-Smith</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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