Secretion of Carboxypeptidase E from Cultured Astrocytes and from AtT‐20 Cells, a Neuroendocrine Cell Line: Implications for Neuropeptide Biosynthesis

Robyn S. Klein, Banasree Das, Lloyd D. Fricker

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

Abstract: Cultured astrocytes have recently been shown to produce certain neuropeptides, as well as neuropeptide processing enzymes. To characterize the secretory pathway in cultured astrocytes, we used the neuropeptide processing enzyme carboxypeptidase E (CPE) as a marker for neuropeptide secretion. Cultured astrocytes and AtT‐20 cells, a mouse pituitary‐derived neuroendocrine cell line, were labeled with [35S]Met for 15 min and then chased with unlabeled Met. CPE was isolated from either medium or cell extracts using a substrate affinity column. The time course of secretion of radiolabeled CPE was significantly different for cultured astrocytes as compared with AtT‐20 cells. CPE was rapidly secreted from the astrocytes after a 30‐min lag time, presumably reflecting transport through the endoplasmic reticulum and Golgi apparatus, followed by constitutive secretion. The secretion of radiolabeled CPE was essentially complete by 2 h. In contrast, only a portion of the radiolabeled CPE was secreted from AtT‐20 cells over a 2–3‐h period, indicating that the majority of newly synthesized CPE is stored, presumably in secretory granules within the AtT‐20 cells. The regulation of CPE secretion from astrocytes was also examined. CPE secretion is stimulated two‐ to threefold by prolonged treatment (3–48 h) with the phorbol ester 12‐O‐tetra‐decanoylphorbol 13‐acetate (TPA) but not by treatment with other secretagogues that stimulate CPE secretion from AtT‐20 cells (forskolin, isoproterenol, A23187, and vasoactive intestinal peptide) or short (<3 h) exposure to TPA. Taken together, these results indicate that the secretory pathway for CPE, and presumably neuropeptides, is substantially different in astrocytes than the secretory pathway for CPE in neuroendocrine cells.

Original languageEnglish (US)
Pages (from-to)2011-2018
Number of pages8
JournalJournal of Neurochemistry
Volume58
Issue number6
DOIs
StatePublished - Jun 1992

Keywords

  • Astrocytes
  • Carboxypeptidases
  • Glia
  • Neuropeptide processing enzymes
  • Neuropeptides
  • Secretory pathway

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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