Expression of hydrophilic surfactant proteins by mesentery cells in rat and man.
Human peritoneal dialysis effluent (PDE) contains a phosphatidylcholine-rich compound similar to the surfactant that lines lung alveoli. This material is secreted by mesothelial cells. Lung surfactant is also characterized by four proteins essential to its function. After having long been considered as lung-specific, some of them have been found in gastric and intestinal epithelial cells. To explore further the similarity between lung and peritoneal surfactants, we investigated whether mesothelial cells also produce surfactant proteins. We used rat transparent mesentery, human visceral peritoneum biopsies and PDE. Surfactant proteins were searched for after one- and two-dimensional SDS/PAGE and Western blotting. On a one-dimensional Western blot, bands at 38 and 66 kDa in rat mesentery, and at 38 and 66 kDa in human peritoneal mesothelial cells (in vivo and in vitro) and PDE, corresponded to monomeric and dimeric forms of lung surfactant protein A (SP-A). On two-dimensional Western blots, the 32 and 38 kDa spots in mesentery and PDE localized at the acidic pH appropriate to the SP-A monomer's isoelectric point. SP-D was also identified at the same 43 kDa molecular mass as in lung. SP-B was not detected in mesenteric samples. Expression of SP mRNA species was also assessed by reverse transcriptase-PCR, which was performed with specific primers of surfactant protein cDNA sequences. With primers of SP-A and SP-D, DNA fragments of the same size were amplified in lung and mesentery, indicating the presence of SP-A and SP-D mRNA species. These fragments were labelled by appropriate probes in a Southern blot. No amplification was obtained for SP-B. These results show that mesentery cells produce SP-A and SP-D, although they are of embryonic origin (mesodermal) and are different from those of the lung and digestive tract (endodermal) that secrete these surfactants.
Chailley-Heu
B
B
INSERM U319, Université Denis Diderot-Paris, France.
Rubio
S
S
Rougier
J P
JP
Ducroc
R
R
Barlier-Mur
A M
AM
Ronco
P
P
Bourbon
J R
JR
D000818:Animals; D015153:Blotting, Western; D018076:DNA, Complementary / Q000032:analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D006023:Glycoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008643:Mesentery / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D011510:Proteolipids / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D037662:Pulmonary Surfactant-Associated Protein A; D037663:Pulmonary Surfactant-Associated Protein D; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D014158:Transcription, Genetic; D014867:Water
D018076:DNA, Complementary; D006023:Glycoproteins; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037663:Pulmonary Surfactant-Associated Protein D; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D014867:Water