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Am J Physiol Lung Cell Mol Physiol 274: L901-L907, 1998;
1040-0605/98 $5.00
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Vol. 274, Issue 6, L901-L907, June 1998

PKC isoforms and other signaling proteins involved in surfactant secretion in developing rat type II cells

Laurice I. Gobran, Zhi-Xin Xu, and Seamus A. Rooney

Division of Perinatal Medicine, Department of Pediatrics, Yale University School of Medicine, New Haven, Connecticut 06510

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 , -beta I, -beta II, -delta , -eta , -zeta , -theta , and -µ, PLC-beta 3, and Gqalpha in adult and newborn type II cells. PKC-epsilon , -gamma , and -lambda and PLC-beta 1, -beta 2, and -beta 4 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 , -beta I, -beta II, 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 -µ, PLC-beta 3, 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 , beta I, beta II, or zeta PKC isoforms.

P2Y2 purinoceptor; adenosine receptors; phospholipase C-beta ; adenosine 5'-triphosphate; Gqalpha ; protein kinase C


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