Chronic intrauterine pulmonary hypertension compromises fetal pulmonary artery smooth muscle cell O2 sensing.
To test the hypothesis that chronic intrauterine pulmonary hypertension (PHTN) compromises pulmonary artery (PA) smooth muscle cell (SMC) O2 sensing, fluorescence microscopy was used to study the effect of an acute increase in Po2 on the cytosolic Ca2+ concentration ([Ca2+]i) of chronically hypoxic subconfluent monolayers of PA SMC in primary culture. PA SMCs were derived from fetal lambs with PHTN due to intrauterine ligation of the ductus arteriosus. Acute normoxia decreased [Ca2+]i in control but not PHTN PA SMC. In control PA SMC, [Ca2+]i increased after Ca2+-sensitive (KCa) and voltage-sensitive (Kv) K+ channel blockade and decreased after diltiazem treatment. In PHTN PA SMC, KCa blockade had no effect, whereas Kv blockade and diltiazem increased [Ca2+]i. Inhibition of sarcoplasmic reticulum Ca2+ ATPase activity caused a greater increase in [Ca2+]i in controls compared with PHTN PA SMC. Conversely, ryanodine caused a greater increase of [Ca2+]i in PHTN compared with control PA SMC. KCa channel mRNA is decreased and Kv channel mRNA is unchanged in PHTN PA SMC compared with controls. We conclude that PHTN compromises PA SMC O2 sensing, alters intracellular Ca2+ homeostasis, and changes the predominant ion channel that determines basal [Ca2+]i from KCa to Kv.
Linden
Bradley C
BC
Department of Surgery, University of Minnesota Medical School, 420 Delaware St. SE, Minneapolis, MN 55455, USA. lind0186@umn.edu
Resnik
Ernesto R
ER
Hendrickson
Kristine J
KJ
Herron
Jean M
JM
O'Connor
Timothy J
TJ
Cornfield
David N
DN
D000818:Animals; D001798:Blood Proteins / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005315:Fetal Diseases / Q000378:metabolism / Q000503:physiopathology*; D005333:Fetus; D006976:Hypertension, Pulmonary / Q000378:metabolism / Q000503:physiopathology*; D000860:Hypoxia / Q000378:metabolism / Q000503:physiopathology; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology*; D010100:Oxygen / Q000494:pharmacology*; D010455:Peptides / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism; D011247:Pregnancy; D011651:Pulmonary Artery / Q000166:cytology / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012433:Ryanodine / Q000494:pharmacology; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D012756:Sheep; D019284:Thapsigargin / Q000494:pharmacology
D001798:Blood Proteins; D004791:Enzyme Inhibitors; D010455:Peptides; D015221:Potassium Channels; D012333:RNA, Messenger; D012433:Ryanodine; D019284:Thapsigargin; C064719:iberiotoxin; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000252:Calcium-Transporting ATPases; D011188:Potassium; D010100:Oxygen; D002118:Calcium