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The Journal of Experimental Biology 205, 415-425 (2002)
© 2002 The Company of Biologists Limited

The pulmonary surfactant system matures upon pipping in the freshwater turtle Chelydra serpentina

Sonya D. Johnston1,2, Christopher B. Daniels1,2, David Cenzato2, Jeffrey A. Whitsett3 and Sandra Orgeig1,2,*

1 Department of Physiology and
2 Department of Environmental Biology,University of Adelaide, Adelaide SA 5005, Australia,
3 Division of Pulmonary Biology and Neonatology, Children’s Hospital Medical Center, Cincinnati OH 45229–3039, USA

*Author for correspondence at address 2 (e-mail: sandra.orgeig{at}adelaide.edu.au)

Accepted 19 November 2001

Pulmonary surfactant (PS), a mixture of phospholipids (PL), neutral lipids and surfactant proteins (SP), lowers surface tension within the lung, which increases lung compliance and improves the removal of fluid at birth. Here, we have examined the expression of thyroid transcription factor-1 (TTF-1) and the surfactant protein SP-B, and also the composition of pulmonary surfactant lipids in the developing lung of the turtle Chelydra serpentina. Lavage and lung tissue were collected from late embryonic, pipped and hatchling turtles. TTF-1, a regulator of gene expression of surfactant proteins and cell differentiation in mammals, was detected using immunohistochemistry in epithelia of the gas-exchange area and conducting airways during late development. Expression declined in hatchlings. SP-B was detected in subsets of cells within the respiratory epithelium at all stages sampled. The same cell types also stained for TTF-1. Turtle surfactant lipids matured toward the end of incubation. Maximal secretion of both total phospholipids and disaturated phospholipid (DSP) occurred at the time of pipping, coincident with the onset of breathing. The DSP/PL ratio increased after pipping, whereas cholesterol levels (Chol) increased prior to pipping. This resulted in a decrease in the Chol/PL and Chol/DSP ratios after pipping. Thus, TTF-1 and SP-B appear to be highly conserved within the vertebrates. Maturation of surfactant phospholipid content occurred with the commencement of pulmonary ventilation.

Key words: thyroid transcription factor-1, surfactant protein, SP-B, phospholipid, cholesterol, lung, development, freshwater turtle, Chelydra serpentina.


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© The Company of Biologists Ltd 2002