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Am. J. Respir. Cell Mol. Biol., Volume 26, Number 6, June, 2002 709-715

Effect Of CTP:Phosphocholine Cytidylyltransferase Overexpression on the Mouse Lung Surfactant System

Jiali Li, James J. Marsh, and Roger G. Spragg

Department of Medicine, University of California, San Diego; and Medicine Service, San Diego Veterans Affairs Healthcare System, San Diego, California

CTP:phosphocholine cytidylyltransferase (CT) is the rate-limiting enzyme in the biosynthesis by type II pneumocytes of phosphatidylcholine (PC), the predominant phospholipid in lung surfactant. Augmentation of endogenous CT activity might therefore result in enhanced surfactant PC production. To test this hypothesis, transgenic mice were created in which rat CT (rCT) was expressed under control of the human surfactant protein C (SP-C) promoter. Transgenic mice were identified by tail-clip PCR analysis and studies of four founder lines were initiated. Lung CT gene expression was enhanced in two transgenic founder lines relative to wild-type controls. These two transgenic lines also exhibited significantly higher levels of immunoreactive CT protein and CT activity in whole-lung homogenates and in cultured type II cell extracts. Disaturated PC (DSPC) content in whole-lung homogenates and the rate of DSPC synthesis in cultured type II cells were significantly increased in one transgenic line. However, neither the incorporation of radiolabeled precursors (choline and palmitate) into DSPC in vivo nor the cellular metabolism of DSPC differed significantly between transgenic and control mice. This transgenic model provides opportunity for further study of factors controlling surfactant phospholipid production in vivo.

Abbreviations: CTP:phosphocholine cytidylyltransferase, CT; Dulbecco's modified Eagle's medium, DMEM; dipalmitoylphosphatidylcholine, DPPC; disaturated phosphatidylcholine, DSPC; disintegrations per min, dpm; Dulbecco's phosphate-buffered saline, PBS; phosphatidylcholine, PC; polymerase chain reaction, PCR; rat CT, rCT; surfactant protein C, SP-C.




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