Methylamine decreases trafficking and packaging of newly synthesized phosphatidylcholine in lamellar bodies in alveolar type II cells.
Lung lamellar bodies, the storage organelles for lung surfactant phosphatidylcholine (PC), maintain an acidic pH that can be increased with weak bases. This study investigates the effect of a weak base, methylamine, on the pH in lamellar bodies and on the trafficking and packaging of newly synthesized PC in lamellar bodies. Methylamine increased the pH of isolated lung lamellar bodies and of lamellar bodies in intact cells. Metabolic labelling of isolated type II cells with [methyl-3H]choline showed that although methylamine (2.5-10 mM) did not alter the labelling of cellular or microsomal PC and disaturated PC, it decreased the labelling of the PC and disaturated PC in lamellar bodies. The packaging of PC in lamellar bodies (the specific activities ratio between the PC in lamellar bodies and the microsomal PC) also decreased in a time- and concentration-dependent manner. The cellular synthesis of PC or its packaging into lamellar bodies was unaltered by brefeldin A, suggesting that the Golgi was not involved in PC packaging. Although methylamine also increased surfactant secretion, the inhibition of PC packaging in lamellar bodies seems unrelated to the secretagogue effect, (1) on the basis of metabolic consequences of increased secretion and (2) because ATP, another secretagogue, did not inhibit PC packaging. Methylamine seems to inhibit PC packaging by inhibiting trafficking of PC to lipid-rich light subcellular fractions. Together our results suggest that the trafficking of surfactant PC into lamellar bodies might be sensitive to changes in the pH of lamellar bodies.
Chander
A
A
Department of Pediatrics, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Sen
N
N
Wu
A M
AM
Higgins
S
S
Wadsworth
S
S
Spitzer
A R
AR
D000165:Acridine Orange / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D020126:Brefeldin A; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002794:Choline / Q000378:metabolism; D003517:Cyclopentanes / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007476:Ionophores / Q000494:pharmacology; D008744:Methylamines / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D008985:Monensin / Q000494:pharmacology; D015388:Organelles / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000096:biosynthesis / Q000378:metabolism*; D010743:Phospholipids / Q000032:analysis; D011650:Pulmonary Alveoli / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011663:Pulmonary Surfactants / Q000378:metabolism; D011796:Quinacrine / Q000378:metabolism; D051381:Rats
D003517:Cyclopentanes; D007476:Ionophores; D008744:Methylamines; D010713:Phosphatidylcholines; D010743:Phospholipids; D011663:Pulmonary Surfactants; D000643:Ammonium Chloride; D020126:Brefeldin A; D000255:Adenosine Triphosphate; D008985:Monensin; C027451:methylamine; D000165:Acridine Orange; D011796:Quinacrine; D002794:Choline