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Department of Ultrastructural Research, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto 606, Japan
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 T1
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.
heparin/heparan sulfate proteoglycan; cell spreading; sodium
chlorate; T1
; surfactant-associated proteins
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