PKC isoforms and other signaling proteins involved in surfactant secretion in developing rat type II cells.
We previously reported that there is a developmental increase in surfactant secretion in response to P2Y2 purinoceptor agonists. UTP does not stimulate secretion in type II cells from 1- or 2-day-old rats; there is a small response to UTP in cells from 4-day-old animals, and the response increases with increasing age thereafter. Second messenger formation in response to P2Y2 agonists has a similar developmental pattern. We have investigated whether the failure to respond to P2Y2 agonists is due to a deficiency in the P2Y2 receptor or in downstream signaling factors. We compared type II cells from adult and 1- to 2-day-old rats with respect to expression of the P2Y2 receptor gene and the levels of phospholipase C-beta (PLC-beta) and protein kinase C (PKC) isomers and of the alpha-subunit of the GTP-binding protein Gq. We measured gene expression by reverse transcriptase-polymerase chain reaction and protein levels by immunoblotting. We identified PKC-alpha, -betaI, -betaII, -delta, -eta, -zeta, -theta, and -mu, PLC-beta3, and Gqalpha in adult and newborn type II cells. PKC-epsilon, -gamma, and -lambda and PLC-beta1, -beta2, and -beta4 were not present in adult or newborn type II cells. Expression of the P2Y2 receptor gene was essentially the same in newborn and adult cells. However, the levels of PKC-alpha, -betaI, -betaII, and -zeta in newborn type II cells were only 43-57% those of adult cells. The level of PKC-theta also tended to be lower in the newborn cells. There was little difference between newborn and adult type II cells in the levels of PKC-delta, -eta, and -mu, PLC-beta3, and Gqalpha. These data suggest that the lack of response of early newborn type II cells to P2Y2 agonists is not due to a lack of expression of the receptor gene but possibly to insufficient amounts of one or more of the alpha, betaI, betaII, or zeta PKC isoforms.
Gobran
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Division of Perinatal Medicine, Department of Pediatrics, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Xu
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Rooney
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SA
D000375:Aging; D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D007527:Isoenzymes / Q000378:metabolism*; D008168:Lung / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism*; D008297:Male; D016133:Polymerase Chain Reaction; D011493:Protein Kinase C / Q000378:metabolism*; D011663:Pulmonary Surfactants / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase; D051381:Rats; D017207:Rats, Sprague-Dawley; D011943:Receptors, Adrenergic, beta / Q000235:genetics; D018048:Receptors, Purinergic P2 / Q000235:genetics; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000378:metabolism; D014544:Uridine Triphosphate / Q000494:pharmacology
D007527:Isoenzymes; D011663:Pulmonary Surfactants; D012333:RNA, Messenger; D011943:Receptors, Adrenergic, beta; D018048:Receptors, Purinergic P2; D011493:Protein Kinase C; D012194:RNA-Directed DNA Polymerase; D010738:Type C Phospholipases; D014544:Uridine Triphosphate