| Am J Respir Cell Mol Biol. 1991 Jun;4(6):489-96. |
|
Identification of pneumocin, a developmentally regulated apical membrane glycoprotein
in rat lung type II and Clara cells.
Lwebuga-Mukasa JS.
Department of Internal Medicine, Yale School of Medicine, New Haven, Connecticut 06510.
Pneumocin (Mr, 165 kD) is a recently identified apical
membrane surface sialoglycoprotein marker of type
II pneumocytes. A murine
monoclonal IgG1 subclass-producing clone 4A (4A
mAb), which was developed against the purified pneumocin, and recognized pneumocin
on Western blots of adult rat lung homogenates, was used to study expression
of the glycoprotein in developing rat lungs. Pneumocin
localized to apical membranes of late fetal, neonatal, and adult rat type
II pneumocytes as well as Clara cells in situ, by immunofluorescence and immunoelectron
microscopy. Faint immunofluorescence was observed
in 17-d fetal lungs. However, 19-d fetal lungs showed intense immunofluorescence
with the antibody. On immunoelectron microscopy,
apical membranes of 19-d fetal and adult rat lung type II cells were labeled
by 4A mAb, but type I cells were not stained. On
Western blots, amounts of pneumocin increased up
to the fourth day after birth, when near-adult levels were attained. Lower
molecular weight forms (Mr, 80 to 90 kD) were recognized in 17-d fetal lung. These bands
decreased in amount with a corresponding increase in the 165-kD band that was typically observed in adult lungs. Immunoglobulins that were eluted from polyvinylidene
difluoride strips containing the 165-kD band recognized the Mr 80 to
90 kD bands and 50-kD component, suggesting that fetal forms of the protein shared
an epitope in common with the adult pneumocin. Reactivity of the glycoprotein with 4A mAb was destroyed by enzymatic digestion with trypsin and staphylococcal V8 protease. These data demonstrate
that pneumocin is a developmentally regulated apical
membrane marker of differentiated type II and Clara cells.(ABSTRACT TRUNCATED AT 250 WORDS)
PMID: 2054191 [PubMed
- indexed for MEDLINE]