Vectorial secretion of perivitelline membrane glycoprotein ZPC of Japanese quail (Coturnix japonica) in polarized Madin-Darby canine kidney cells.
The avian perivitelline membrane (PL), which is an investment homologous to the mammalian zona pellucida, is found between the surface of the oocyte and the apical surface of ovarian granulosa cells. Our previous study demonstrated that ZPC, one of the components of PL, is synthesized in ovarian granulosa cells. However, how the secretion of ZPC is regulated in the cells has been insufficiently investigated. We studied the secretion of quail ZPC expressed in polarized Madin-Darby canine kidney (MDCK) cells in a dual-chamber apparatus. Western blot analyses of the conditioned medium demonstrated that the majority of the secreted ZPC were distributed in the apical compartment. When ZPC lacking N-linked oligosaccharides was transfected into the cells, the 31-kDa immunoreactive band was detected in both the apical and the basolateral medium. Interestingly, immunohistochemical observations of the follicular wall demonstrated that the predominant intracellular form of ZPC in the cells localized in the apical side of the perinuclear region apposed to the PL, but not the basolateral side, indicating the possibility that ZPC could be selectively transported toward the apical surface in vivo. Taken together, these results indicated that ZPC expressed in MDCK cells are selectively released to the apical compartment, and that the N-linked carbohydrates might possess information that causes the efficient transport of ZPC to the apical surface of the cells.
Sasanami
Tomohiro
T
Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Shizuoka, Japan. atsasan@agr.shizuoka.ac.jp
Matsushima
Kayoko
K
Ohtsuki
Mamoru
M
Kansaku
Norio
N
Hiyama
Gen
G
Mori
Makoto
M
D000818:Animals; D030161:Avian Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D003370:Coturnix; D017077:Culture Media, Conditioned; D004285:Dogs; D004527:Egg Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D007668:Kidney / Q000166:cytology*; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology; D014817:Vitelline Membrane / Q000737:chemistry*
D030161:Avian Proteins; D017077:Culture Media, Conditioned; D004527:Egg Proteins; D008562:Membrane Glycoproteins