Cytochemical characterization of anterior pituitary target cells for the neuropeptide, pituitary adenylate cyclase activating polypeptide (PACAP), using biotinylated ligands

Sandor Vigh, Akira Arimura, Paul E. Gottschall, Chieko Kitada, Anikó Somogyvári-Vigh, Gwen V. Childs

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

Two novel peptides, named PACAP (pituitary adenylate cyclase activating polypeptide) containing 38 (PACAP38) and 27 residues (PACAP27) were recently isolated from ovine hypothalami. In order to investigate the pituitary cell type(s) that bear a receptor for PACAP, PACAP38 was biotinylated and used for cytochemical examination of binding. The cells were also identified by immunocytochemical methods using the antisera against each of the rat anterior pituitary hormones or an antiserum against S-100 protein, a marker for pituitary folliculo-stellate (FS) cells. Biotinylated PACAP38 (biot-PACAP) exhibited adenylate cyclase stimulating activity (ACSA) comparable to PACAP38 in rat pituitary cell cultures, and displaced the bound 125I-PACAP27 to the rat pituitary membrane preparation to the same extent as PACAP38. After 2-4 days of culture, dispersed rat pituitary cells were incubated with varying concentrations of biot-PACAP at room temperature or 4°C. The bound biot-PACAP38 was visualized by avidin-biotin-peroxidase complex (ABC) method with nickel intensification. Biot-PACAP-positive and pituitary hormone or S-100-positive cells were counted. More than 90% of S-100-positive cells bound biot-PACAP38. A considerable number of GH and PRL cells and a lesser number of ACTH cells also bound biot-PACAP38, whereas only a few identified LH, FSH, or TSH cells bound biot-PACAP38. These results suggest that FS cells are a major target cell type for PACAP. A recent study from our laboratory demonstrated that PACAP stimulated the release of interleukin (IL)-6 in rat pituitary cell cultures. FS cells are known to produce IL-6.

Original languageEnglish
Pages (from-to)59-65
Number of pages7
JournalPeptides
Volume14
Issue number1
DOIs
Publication statusPublished - 1993

Fingerprint

Pituitary Adenylate Cyclase-Activating Polypeptide
Neuropeptides
Ligands
Rats
Gonadotrophs
Cell culture
Immune Sera
Interleukin-6
Pituitary Adenylate Cyclase-Activating Polypeptide Receptors
Cells
Cell Culture Techniques
Anterior Pituitary Hormones
Thyrotrophs
Pituitary Hormones
Avidin
S100 Proteins
Peptide Hormones
Biotin
Nickel
Adenylyl Cyclases

Keywords

  • Biotin labeled
  • Biotinylated ligand cytochemistry
  • Folliculo-stellate cells
  • Immunocytochemistry
  • PACAP
  • Rat pituitary
  • Receptors
  • Target cells

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Physiology
  • Cellular and Molecular Neuroscience

Cite this

Cytochemical characterization of anterior pituitary target cells for the neuropeptide, pituitary adenylate cyclase activating polypeptide (PACAP), using biotinylated ligands. / Vigh, Sandor; Arimura, Akira; Gottschall, Paul E.; Kitada, Chieko; Somogyvári-Vigh, Anikó; Childs, Gwen V.

In: Peptides, Vol. 14, No. 1, 1993, p. 59-65.

Research output: Contribution to journalArticle

Vigh, Sandor ; Arimura, Akira ; Gottschall, Paul E. ; Kitada, Chieko ; Somogyvári-Vigh, Anikó ; Childs, Gwen V. / Cytochemical characterization of anterior pituitary target cells for the neuropeptide, pituitary adenylate cyclase activating polypeptide (PACAP), using biotinylated ligands. In: Peptides. 1993 ; Vol. 14, No. 1. pp. 59-65.
@article{2ba2764ecc5f4a6590ff8ad0649612ed,
title = "Cytochemical characterization of anterior pituitary target cells for the neuropeptide, pituitary adenylate cyclase activating polypeptide (PACAP), using biotinylated ligands",
abstract = "Two novel peptides, named PACAP (pituitary adenylate cyclase activating polypeptide) containing 38 (PACAP38) and 27 residues (PACAP27) were recently isolated from ovine hypothalami. In order to investigate the pituitary cell type(s) that bear a receptor for PACAP, PACAP38 was biotinylated and used for cytochemical examination of binding. The cells were also identified by immunocytochemical methods using the antisera against each of the rat anterior pituitary hormones or an antiserum against S-100 protein, a marker for pituitary folliculo-stellate (FS) cells. Biotinylated PACAP38 (biot-PACAP) exhibited adenylate cyclase stimulating activity (ACSA) comparable to PACAP38 in rat pituitary cell cultures, and displaced the bound 125I-PACAP27 to the rat pituitary membrane preparation to the same extent as PACAP38. After 2-4 days of culture, dispersed rat pituitary cells were incubated with varying concentrations of biot-PACAP at room temperature or 4°C. The bound biot-PACAP38 was visualized by avidin-biotin-peroxidase complex (ABC) method with nickel intensification. Biot-PACAP-positive and pituitary hormone or S-100-positive cells were counted. More than 90{\%} of S-100-positive cells bound biot-PACAP38. A considerable number of GH and PRL cells and a lesser number of ACTH cells also bound biot-PACAP38, whereas only a few identified LH, FSH, or TSH cells bound biot-PACAP38. These results suggest that FS cells are a major target cell type for PACAP. A recent study from our laboratory demonstrated that PACAP stimulated the release of interleukin (IL)-6 in rat pituitary cell cultures. FS cells are known to produce IL-6.",
keywords = "Biotin labeled, Biotinylated ligand cytochemistry, Folliculo-stellate cells, Immunocytochemistry, PACAP, Rat pituitary, Receptors, Target cells",
author = "Sandor Vigh and Akira Arimura and Gottschall, {Paul E.} and Chieko Kitada and Anik{\'o} Somogyv{\'a}ri-Vigh and Childs, {Gwen V.}",
year = "1993",
doi = "10.1016/0196-9781(93)90011-5",
language = "English",
volume = "14",
pages = "59--65",
journal = "Peptides",
issn = "0196-9781",
publisher = "Elsevier Inc.",
number = "1",

}

TY - JOUR

T1 - Cytochemical characterization of anterior pituitary target cells for the neuropeptide, pituitary adenylate cyclase activating polypeptide (PACAP), using biotinylated ligands

AU - Vigh, Sandor

AU - Arimura, Akira

AU - Gottschall, Paul E.

AU - Kitada, Chieko

AU - Somogyvári-Vigh, Anikó

AU - Childs, Gwen V.

PY - 1993

Y1 - 1993

N2 - Two novel peptides, named PACAP (pituitary adenylate cyclase activating polypeptide) containing 38 (PACAP38) and 27 residues (PACAP27) were recently isolated from ovine hypothalami. In order to investigate the pituitary cell type(s) that bear a receptor for PACAP, PACAP38 was biotinylated and used for cytochemical examination of binding. The cells were also identified by immunocytochemical methods using the antisera against each of the rat anterior pituitary hormones or an antiserum against S-100 protein, a marker for pituitary folliculo-stellate (FS) cells. Biotinylated PACAP38 (biot-PACAP) exhibited adenylate cyclase stimulating activity (ACSA) comparable to PACAP38 in rat pituitary cell cultures, and displaced the bound 125I-PACAP27 to the rat pituitary membrane preparation to the same extent as PACAP38. After 2-4 days of culture, dispersed rat pituitary cells were incubated with varying concentrations of biot-PACAP at room temperature or 4°C. The bound biot-PACAP38 was visualized by avidin-biotin-peroxidase complex (ABC) method with nickel intensification. Biot-PACAP-positive and pituitary hormone or S-100-positive cells were counted. More than 90% of S-100-positive cells bound biot-PACAP38. A considerable number of GH and PRL cells and a lesser number of ACTH cells also bound biot-PACAP38, whereas only a few identified LH, FSH, or TSH cells bound biot-PACAP38. These results suggest that FS cells are a major target cell type for PACAP. A recent study from our laboratory demonstrated that PACAP stimulated the release of interleukin (IL)-6 in rat pituitary cell cultures. FS cells are known to produce IL-6.

AB - Two novel peptides, named PACAP (pituitary adenylate cyclase activating polypeptide) containing 38 (PACAP38) and 27 residues (PACAP27) were recently isolated from ovine hypothalami. In order to investigate the pituitary cell type(s) that bear a receptor for PACAP, PACAP38 was biotinylated and used for cytochemical examination of binding. The cells were also identified by immunocytochemical methods using the antisera against each of the rat anterior pituitary hormones or an antiserum against S-100 protein, a marker for pituitary folliculo-stellate (FS) cells. Biotinylated PACAP38 (biot-PACAP) exhibited adenylate cyclase stimulating activity (ACSA) comparable to PACAP38 in rat pituitary cell cultures, and displaced the bound 125I-PACAP27 to the rat pituitary membrane preparation to the same extent as PACAP38. After 2-4 days of culture, dispersed rat pituitary cells were incubated with varying concentrations of biot-PACAP at room temperature or 4°C. The bound biot-PACAP38 was visualized by avidin-biotin-peroxidase complex (ABC) method with nickel intensification. Biot-PACAP-positive and pituitary hormone or S-100-positive cells were counted. More than 90% of S-100-positive cells bound biot-PACAP38. A considerable number of GH and PRL cells and a lesser number of ACTH cells also bound biot-PACAP38, whereas only a few identified LH, FSH, or TSH cells bound biot-PACAP38. These results suggest that FS cells are a major target cell type for PACAP. A recent study from our laboratory demonstrated that PACAP stimulated the release of interleukin (IL)-6 in rat pituitary cell cultures. FS cells are known to produce IL-6.

KW - Biotin labeled

KW - Biotinylated ligand cytochemistry

KW - Folliculo-stellate cells

KW - Immunocytochemistry

KW - PACAP

KW - Rat pituitary

KW - Receptors

KW - Target cells

UR - http://www.scopus.com/inward/record.url?scp=0027450080&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027450080&partnerID=8YFLogxK

U2 - 10.1016/0196-9781(93)90011-5

DO - 10.1016/0196-9781(93)90011-5

M3 - Article

C2 - 8441707

AN - SCOPUS:0027450080

VL - 14

SP - 59

EP - 65

JO - Peptides

JF - Peptides

SN - 0196-9781

IS - 1

ER -