Differential Regulation of Myosin Regulatory Light Chain Phosphorylation by Protein Kinase C Isozymes in Human Uterine Myocytes.
Animals
Cells, Cultured
Enzyme Activation
Enzyme Activators
/ pharmacology
Female
Humans
Intracellular Signaling Peptides and Proteins
/ metabolism
Isoenzymes
Muscle Proteins
/ metabolism
Myocytes, Smooth Muscle
/ drug effects
Myosin Light Chains
/ metabolism
Myosin-Light-Chain Kinase
/ metabolism
Oxytocics
/ pharmacology
Phosphorylation
Protein Kinase C
/ antagonists & inhibitors
Protein Kinase Inhibitors
/ pharmacology
Rats, Sprague-Dawley
Signal Transduction
Uterine Contraction
/ drug effects
Uterus
/ cytology
CPI-17
myosin regulatory light chains
parturition
preterm labor
protein kinase C
uterine contractility
Journal
Reproductive sciences (Thousand Oaks, Calif.)
ISSN: 1933-7205
Titre abrégé: Reprod Sci
Pays: United States
ID NLM: 101291249
Informations de publication
Date de publication:
07 2019
07 2019
Historique:
pubmed:
16
11
2018
medline:
14
4
2020
entrez:
16
11
2018
Statut:
ppublish
Résumé
Preterm birth is the most common cause of neonatal morbidity and mortality and a common precedent to lifelong disability. Current treatment has minimal efficacy. We assessed the role of isozymes of the protein kinase C (PKC) family in regulating the phosphorylation of myosin regulatory light chains (RLCs), which regulate uterine contractility. We also explored the mechanisms through which these isozymes function. We used a previously characterized and validated quantitative in-cell Western (ICW) assay to measure site-specific phosphorylations on myosin RLC and CPI-17. Cultures of human uterine myocytes (hUM) were treated with the potent contractile stimulant oxytocin to induce uterine contractility or a pharmacological mimic of diacyl-glycerol to stimulate the conventional and novel isozymes of the PKC family. Combinations of isozyme-selective inhibitors were used to determine the effects of the conventional and novel classes of isozymes. Stimulation of PKC using phospho-dibutyrate caused immediate, concentration-dependent inhibition of uterine activity Specific agonists for the conventional isozymes or inhibitors of the novel isozymes of the PKC family could be useful pharmacological agents for regulation of uterine activity.
Sections du résumé
BACKGROUND
Preterm birth is the most common cause of neonatal morbidity and mortality and a common precedent to lifelong disability. Current treatment has minimal efficacy.
OBJECTIVE
We assessed the role of isozymes of the protein kinase C (PKC) family in regulating the phosphorylation of myosin regulatory light chains (RLCs), which regulate uterine contractility. We also explored the mechanisms through which these isozymes function.
STUDY DESIGN
We used a previously characterized and validated quantitative in-cell Western (ICW) assay to measure site-specific phosphorylations on myosin RLC and CPI-17. Cultures of human uterine myocytes (hUM) were treated with the potent contractile stimulant oxytocin to induce uterine contractility or a pharmacological mimic of diacyl-glycerol to stimulate the conventional and novel isozymes of the PKC family. Combinations of isozyme-selective inhibitors were used to determine the effects of the conventional and novel classes of isozymes.
RESULTS
Stimulation of PKC using phospho-dibutyrate caused immediate, concentration-dependent inhibition of uterine activity
CONCLUSIONS
Specific agonists for the conventional isozymes or inhibitors of the novel isozymes of the PKC family could be useful pharmacological agents for regulation of uterine activity.
Identifiants
pubmed: 30428777
doi: 10.1177/1933719118802062
pii: 10.1177/1933719118802062
doi:
Substances chimiques
Enzyme Activators
0
Intracellular Signaling Peptides and Proteins
0
Isoenzymes
0
Muscle Proteins
0
Myosin Light Chains
0
Oxytocics
0
PPP1R14A protein, human
0
Protein Kinase Inhibitors
0
Protein Kinase C
EC 2.7.11.13
Myosin-Light-Chain Kinase
EC 2.7.11.18
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
988-996Subventions
Organisme : CIHR
Pays : Canada
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March of Dimes, The Partnership for Maternal, Newborn & Child Health, Save the Children, World Health Organization. Born Too Soon: The Global Action Report on Preterm Birth. Geneva: World Health Organization; 2012.
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