title: The secretory pathway is blocked between the trans-Golgi and the plasma membrane during meiotic maturation in Xenopus oocytes.
abstract: Protein secretion is blocked in Xenopus oocytes arrested at second meiotic metaphase. In this report, we show that secretion becomes blocked coincident with germinal vesicle breakdown (GVBD). Transport through the metaphase-arrested oocyte's secretory pathway continues unimpeded until proteins reach the trans-Golgi. These conclusions are drawn from experiments using exogenous prolactin and vesicular stomatitis virus G protein (VSV G) encoded by SP6 transcripts and endogenous glycosaminoglycan (GAG) chains initiated on beta-D-4-methylumbelliferyl-xyloside. From the initiation of maturation with progesterone until GVBD, secretion of prolactin synthesized before the start of maturation is comparable to secretion in immature oocytes, but after GVBD secretion of prolactin declines approximately 63% in the first hour. Not all steps in the secretory pathway are blocked when oocytes mature. Since VSV G protein acquires resistance to endo H digestion with equal efficiency in immature oocytes (arrested in first meiotic prophase) and matured oocytes (arrested in second meiotic metaphase), we conclude that transport of this protein from the ER to the Golgi is not inhibited at meiotic metaphase. Using [35S]sulfate to label xyloside-initiated GAG chains we find that transport of GAG chains from the trans-Golgi to the cell surface is 15-fold lower in matured oocytes than in immature oocytes. Examination of the size of GAG chains by SDS-PAGE and HPLC indicates that matured oocytes produce GAG chains significantly larger than GAG chains from immature oocytes. This increase in size suggests that GAG chains from matured oocytes have a longer residence time in the trans-Golgi than GAG chains from immature oocytes. Hence, part of the block to secretion in metaphase-arrested oocytes could be an inhibition of vesicle budding from the trans-Golgi.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley 94720.
authors.forename: D S
authors.initials: DS
authors.lastname: Leaf
authors.forename: S J
authors.initials: SJ
authors.lastname: Roberts
authors.forename: J C
authors.initials: JC
authors.lastname: Gerhart
authors.forename: H P
authors.initials: HP
authors.lastname: Moore
mesh_terms: D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid; D004721:Endoplasmic Reticulum / Q000502:physiology; D006056:Golgi Apparatus / Q000502:physiology* / Q000648:ultrastructure; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000502:physiology*; D008540:Meiosis / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000493:pharmacokinetics; D009865:Oocytes / Q000378:metabolism*; D011233:Precipitin Tests; D011374:Progesterone / Q000494:pharmacology; D011388:Prolactin / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000493:pharmacokinetics; D014982:Xenopus laevis
chemical_list: C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D011374:Progesterone; D011388:Prolactin