Expression of N-deacetylase/sulfotransferase and 3-O-sulfotransferase in rat alveolar type II cells.
Basal laminae beneath alveolar type I cells are suggested to contain highly sulfated heparan sulfate-containing proteoglycans (PGs), and cultured type II cells accumulate highly sulfated matrices. To characterize the regulation of PG synthesis during the transition from type II cells to type I cells, we examined mRNA expression of N-deacetylase/sulfotransferase (NST) and 3-O-sulfotransferase (3-OST), two enzymes specific for heparan sulfate synthesis. We found that both freshly isolated and cultured type II cells expressed NST and 3-OST as shown by in situ hybridization. Expression of surfactant-associated protein A, B, and C mRNAs, determined by semiquantitative PCR, decreased during culture. Expression of type I cell marker T1alpha mRNA increased except in cells cultured on an Engelbrecht-Holm-Swarm gel. Expression of NST was dependent on cell density and matrix and was intense in conditions where cells spread fully, whereas 3-OST expression was unchanged in the conditions examined. The PG sulfation inhibitor sodium chlorate significantly inhibited cultured type II cell spreading, and this inhibition was reversed by sodium sulfate. These results suggest that highly sulfated PGs modified by NST are necessary for the spreading of cells during transdifferentiation of type II cells to mature type I cells.
Department of Ultrastructural Research, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto 606, Japan.
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Li
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Hirayoshi
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Suzuki
D000581:Amidohydrolases / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002452:Cell Count; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002704:Chlorates / Q000494:pharmacology; D003001:Cloning, Molecular; D046508:Culture Techniques / Q000379:methods; D018076:DNA, Complementary / Q000032:analysis*; D004305:Dose-Response Relationship, Drug; D017403:In Situ Hybridization; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D011509:Proteoglycans / Q000096:biosynthesis; D011650:Pulmonary Alveoli / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015238:Sulfotransferases / Q000096:biosynthesis / Q000235:genetics*; D053667:Syndecans
D002704:Chlorates; D018076:DNA, Complementary; D008562:Membrane Glycoproteins; D011509:Proteoglycans; D012333:RNA, Messenger; D053667:Syndecans; D015238:Sulfotransferases; C102180:heparan sulfate D-glucosaminyl 3-O-sulfotransferase; C021350:heparitin sulfotransferase; D000581:Amidohydrolases; C032706:sodium chlorate