title: Neuroserpin is expressed in the pituitary and adrenal glands and induces the extension of neurite-like processes in AtT-20 cells.
abstract: Two cDNAs encoding the serine protease inhibitor (serpin) neuroserpin were cloned from a rat pituitary cDNA library (rNS-1, 2922 bp; rNS-2, 1599 bp). In situ hybridization histochemistry showed neuroserpin transcripts in the intermediate, anterior and posterior lobes of the pituitary gland and medullary cells in the adrenal gland. Expression of rNS-1 mRNA was restricted to selected cells in the pituitary gland. Analysis of purified secretory-granule fractions from pituitary and adrenal tissues indicated that neuroserpin was found in dense-cored secretory granules. This result suggested that endocrine neuroserpin may regulate intragranular proteases or inhibit enzymes following regulated secretion. To investigate the function of neuroserpin in endocrine tissues we established stable anterior pituitary AtT-20 cell lines expressing neuroserpin. Cells with increased levels of neuroserpin responded by extending neurite-like processes. Extracellular proteolysis by serine protease plasminogen activators has been suggested to regulate neurite outgrowth. As neuroserpin inhibits tissue plasminogen activator (tPA) in vitro, we measured plasminogen-activator levels. Zymographic analysis indicated that AtT-20 cells synthesized and secreted a plasminogen activator identical in size to tPA. A higher-molecular-mass tPA-neuroserpin complex was also observed in AtT-20-cell conditioned culture medium. tPA levels were similar in parent AtT-20 cells and a stable cell line with increased levels of neuroserpin. There was no accumulation of a tPA-neuroserpin complex. Together these results identify endocrine cells as an important source of neuroserpin. Moreover they suggest that neuroserpin is released from dense-cored secretory granules to regulate cell-extracellular matrix interactions through a mechanism that may not directly involve tPA.
authors.affiliation: Molecular Neuroendocrinology Laboratory, School of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland, New Zealand.
authors.forename: R M
authors.initials: RM
authors.lastname: Hill
authors.forename: P K
authors.initials: PK
authors.lastname: Parmar
authors.forename: L C
authors.initials: LC
authors.lastname: Coates
authors.forename: E
authors.initials: E
authors.lastname: Mezey
authors.forename: J F
authors.initials: JF
authors.lastname: Pearson
authors.forename: N P
authors.initials: NP
authors.lastname: Birch
mesh_terms: D000311:Adrenal Glands / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D003594:Cytoplasmic Granules / Q000378:metabolism; D008969:Molecular Sequence Data; D016501:Neurites / Q000378:metabolism*; D009479:Neuropeptides / Q000235:genetics* / Q000378:metabolism*; D010902:Pituitary Gland / Q000378:metabolism*; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000378:metabolism; D051381:Rats; D015842:Serine Proteinase Inhibitors / Q000235:genetics / Q000378:metabolism; D015843:Serpins / Q000235:genetics* / Q000378:metabolism*; D013347:Subcellular Fractions; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors / Q000378:metabolism; D000097522:Neuroserpin
chemical_list: D009479:Neuropeptides; D015842:Serine Proteinase Inhibitors; D015843:Serpins; D010959:Tissue Plasminogen Activator; D000097522:Neuroserpin