title: Pituitary adenylate cyclase-activating peptide induces neurite outgrowth in cultured monkey trigeminal ganglion cells: involvement of receptor PAC1.
abstract: OBJECTIVE
  Our previous studies in the rabbit trigeminal nerve (TgN) showed that pituitary adenylate cyclase-activating peptide (PACAP) accelerated the extension of neuronal processes and recovery of corneal sensitivity. The purposes of the present study were 1) develop a procedure to culture trigeminal nerve (TgN) cells from monkeys, 2) test whether PACAP induces sprouting and elongation of axons in our culture system, 3) investigate the signaling mechanisms producing axon elongation induced by PACAP, and 4) test the action of PACAP on tear protein secretion by monkey lacrimal acinar cells.
  
  
  METHODS
  Primary cultures of TgN cells were established from rhesus monkeys. Cellular distribution of the PACAP receptor, PAC1, was determined with immunostaining. Axonal length in cultured TgN ganglion cells was evaluated with staining by antibody for neurofilament. mRNA expression was determined with quantitative real-time polymerase chain reaction (qPCR). Secretion of tear protein from cultured acinar cells was measured with immunoblotting.
  
  
  RESULTS
  Our results showed that dissociated, cultured TgN cells contained neuronal ganglion and Schwann cells, and the PAC1 receptor was expressed in both cell types. PACAP-27 significantly induced neurite outgrowth, which was inhibited by PACAP 6-27. Inhibitors for adenylate cyclase and phospholipase C also inhibited neurite outgrowth. Follistatin was upregulated by PACAP-27 during the culture period. PACAP enhanced secretion of tear proteins.
  
  
  CONCLUSIONS
  Our data suggested PAC1 activation is involved in TgN neurite outgrowth.
authors: Senju Laboratory of Ocular Sciences, Senju Pharmaceutical Corporation Limited, Beaverton, OR, USA.
authors: Emi
authors: E
authors: Nakajima
authors: Ryan D
authors: RD
authors: Walkup
authors: Atsuko
authors: A
authors: Fujii
authors: Thomas R
authors: TR
authors: Shearer
authors: Mitsuyoshi
authors: M
authors: Azuma
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005136:Eye Proteins / Q000378:metabolism; D007765:Lacrimal Apparatus / Q000378:metabolism; D008253:Macaca mulatta; D016501:Neurites / Q000378:metabolism* / Q000648:ultrastructure; D010446:Peptide Fragments / Q000378:metabolism; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D051237:Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I / Q000235:genetics / Q000378:metabolism*; D051239:Receptors, Vasoactive Intestinal Peptide, Type II / Q000235:genetics / Q000378:metabolism; D051238:Receptors, Vasoactive Intestinal Polypeptide, Type I / Q000235:genetics / Q000378:metabolism; D012583:Schwann Cells / Q000378:metabolism; D015398:Signal Transduction; D012668:Trigeminal Ganglion / Q000166:cytology* / Q000378:metabolism*
chemical_list: D005136:Eye Proteins; D010446:Peptide Fragments; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D012333:RNA, Messenger; D051237:Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I; D051239:Receptors, Vasoactive Intestinal Peptide, Type II; D051238:Receptors, Vasoactive Intestinal Polypeptide, Type I; C106317:pituitary adenylate-cyclase-activating peptide (6-27); C042434:tear proteins
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