Secretion of egg white proteins in primary cultured oviduct cells of laying Japanese quail (Coturnix coturnix japonica).
Oviduct (magnum) cells of laying Japanese quails (Coturnix coturnix japonica) were isolated after digestion of the magnum portion by collagenase and dispase and cultured in Dulbecco's modified Eagle medium (DME) supplemented with 10% fetal calf serum. The cultured cells formed a monolayer. Immunoperoxidase staining with antiovalbumin and anticonalbumin antibodies elucidated that more than 90% of the total population of the cells contained ovalbumin and conalbumin within the cytoplasm. The result shows that the cultured cells are composed mainly of tubular gland cells. Cells secreted ovalbumin and conalbumin continuously for several days. Secretion of these egg white proteins was confirmed to be inhibited by colchicine and vinblastine, which means that the microtubular system is involved in the secretion pathway of the proteins, as is widely accepted. Tunicamycin, an inhibitor of glycosylation of proteins, blocked the glycosylation of nascent ovalbumin molecules at a concentration of 5 micrograms/mL in the medium. However, immunoreactive ovalbumin was secreted under these conditions, without being glycosylated. This suggests that the carbohydrate moiety of ovalbumin is not essential for the secretion of ovalbumin. The secretion of ovalbumin was inhibited by brefeldin A, which is supposed to inhibit the transport of proteins from the rough endoplasmic reticulum to the Golgi apparatus. The present method of cultivating egg white-secreting cells will be useful in investigations of mechanisms and regulation of the synthesis and secretion of egg white proteins in birds.
Kato
S
S
Department of Agricultural Chemistry, Faculty of Agriculture, University of Tokyo, Japan.
Noguchi
T
T
Naito
H
H
D000818:Animals; D002478:Cells, Cultured; D003207:Conalbumin / Q000378:metabolism; D003370:Coturnix / Q000502:physiology*; D004527:Egg Proteins / Q000378:metabolism*; D005260:Female; D010047:Ovalbumin / Q000378:metabolism; D010057:Oviducts / Q000166:cytology* / Q000378:metabolism; D011784:Quail / Q000502:physiology*
D004527:Egg Proteins; D003207:Conalbumin; D010047:Ovalbumin