Acute hypoxia causes membrane depolarization and calcium influx in fetal pulmonary artery smooth muscle cells.
Changes in oxygen tension in the perinatal period contribute to high pulmonary vascular tone in the fetus and the decline in resistance that occurs at birth. Distal pulmonary artery smooth muscle cells (PASMC) isolated from late-gestation ovine fetuses respond to acute hypoxia with an increase in cytosolic calcium concentration ([Ca2+]i) dependent on Ca2+ entry. The purpose of this study is to determine 1) whether acute hypoxia results in PASMC membrane depolarization, 2) whether Ca2+ entry was through voltage-operated calcium channels (VOCC), 3) the contribution of Ca(2+)-induced Ca2+ release (CICR) to the hypoxic response, and 4) whether a subset of K+ channels might serve as oxygen sensors in fetal PASMC. We used microfluorimetry on subconfluent monolayers of PASMC in primary culture loaded with either a membrane potential-sensitive dye, bis(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC4; DPASMC), to estimate membrane potential, or the Ca(2+)-sensitive fluorophore, fura 2, to measure [Ca2+]i. Hypoxia increased fluorescence from PASMC loaded with DiBAC4, consistent with membrane depolarization. Verapamil (an inhibitor of VOCC) attenuated, and BAY K 8644 (a VOCC facilitator) potentiated, the hypoxia-induced increase in [Ca2+]i, respectively. The hypoxic response was transient after treatment with ryanodine (10(-7) M), a blocker of calcium release from intracellular stores. Charybdotoxin (10(-7) M), an inhibitor of Ca(2+)-activated K+ channels, almost doubled [Ca2+]i, whereas glibenclamide (10(-5) M), an ATP-sensitive K(+)-channel antagonist, had no effect.(ABSTRACT TRUNCATED AT 250 WORDS)
Cornfield
D N
DN
Department of Pediatrics, Children's Hospital, Denver, Colorado.
Stevens
T
T
McMurtry
I F
IF
Abman
S H
SH
Rodman
D M
DM
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000208:Acute Disease; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D004594:Electrophysiology; D005333:Fetus / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D005453:Fluorescence; D000860:Hypoxia / Q000503:physiopathology*; D008345:Manganese / Q000378:metabolism; D009130:Muscle, Smooth / Q000196:embryology*; D011651:Pulmonary Artery / Q000196:embryology*; D012433:Ryanodine / Q000494:pharmacology; D012756:Sheep; D014700:Verapamil / Q000494:pharmacology
D012433:Ryanodine; D008345:Manganese; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D014700:Verapamil; D002118:Calcium