Peptides bind to eosinophils in the rat stomach

David H. Nichols, Sándor Lovas, Thomas E. Adrian, Caroline A. Miller, Richard F. Murphy

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background: An immunological role for eosinophils has been well established. However, roles for eosinophils in the physiological functions of the organs they populate are little explored. Methods: Fixed, frozen, then vibratomed sections of rat stomach were exposed to biotinylated 1-17 gastrin (bG17), biotinylated gastrin-releasing peptide (bGRP), biotinylated neuromedin C (bNC), biotinylated vasoactive intestinal peptide (bVIP), and biotinylated substance P (bSP). Binding sites were identified using an avidin-biotin-glucose oxidase complex and tetranitroblue tetrazolium staining. Results: bG17, bGRP, and bNC all bound to cells in the lamina propria and to a lesser extent in the submucosa. Neither bVIP nor bSP bound to cells in these sections. Stained cells were identified as eosinophils in the light microscope on the basis of their distribution and staining properties using the Luna stain for eosinophils and in the transmission electron microscope (TEM) on the basis of a light/TEM matching process. Plastic sections viewed in the light microscope showed that stain was localized to a granular component in the cytoplasm of the eosinophils. No other cell type, specifically neither mast cells nor plasma cells, stained. G17 competed for the bG17 binding site better than did NC. A competition study in which polyglutamic acid failed to compete with bG17 for the binding site, and the observation that bG17, bGRP, and bNC did not bind to other positively charged sites (e.g., collagen, red blood corpuscles), demonstrated that binding was not due to nonspecific electrostatic interactions alone. Binding of bG17 to a CCK(B)/gastrin-type receptor was ruled out when specific receptor antagonists failed to block binding. Conclusions: The particulate nature of the binding site suggests a secretory substance. If so, eosinophils might use that substance to destroy, neutralize, or control the activity of peptide hormones bound to it in the extracellular space.

Original languageEnglish
Pages (from-to)172-181
Number of pages10
JournalAnatomical Record
Volume250
Issue number2
DOIs
StatePublished - Feb 1998

Fingerprint

eosinophils
Eosinophils
peptide
Stomach
stomach
gastrin-releasing peptide
peptides
Gastrin-Releasing Peptide
Peptides
binding sites
rats
Binding Sites
Cholecystokinin B Receptor
gastrins
vasoactive intestinal peptide
substance P
transmission electron microscopes
Vasoactive Intestinal Peptide
light microscopes
Substance P

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences (miscellaneous)
  • Anatomy

Cite this

Peptides bind to eosinophils in the rat stomach. / Nichols, David H.; Lovas, Sándor; Adrian, Thomas E.; Miller, Caroline A.; Murphy, Richard F.

In: Anatomical Record, Vol. 250, No. 2, 02.1998, p. 172-181.

Research output: Contribution to journalArticle

Nichols, David H. ; Lovas, Sándor ; Adrian, Thomas E. ; Miller, Caroline A. ; Murphy, Richard F. / Peptides bind to eosinophils in the rat stomach. In: Anatomical Record. 1998 ; Vol. 250, No. 2. pp. 172-181.
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abstract = "Background: An immunological role for eosinophils has been well established. However, roles for eosinophils in the physiological functions of the organs they populate are little explored. Methods: Fixed, frozen, then vibratomed sections of rat stomach were exposed to biotinylated 1-17 gastrin (bG17), biotinylated gastrin-releasing peptide (bGRP), biotinylated neuromedin C (bNC), biotinylated vasoactive intestinal peptide (bVIP), and biotinylated substance P (bSP). Binding sites were identified using an avidin-biotin-glucose oxidase complex and tetranitroblue tetrazolium staining. Results: bG17, bGRP, and bNC all bound to cells in the lamina propria and to a lesser extent in the submucosa. Neither bVIP nor bSP bound to cells in these sections. Stained cells were identified as eosinophils in the light microscope on the basis of their distribution and staining properties using the Luna stain for eosinophils and in the transmission electron microscope (TEM) on the basis of a light/TEM matching process. Plastic sections viewed in the light microscope showed that stain was localized to a granular component in the cytoplasm of the eosinophils. No other cell type, specifically neither mast cells nor plasma cells, stained. G17 competed for the bG17 binding site better than did NC. A competition study in which polyglutamic acid failed to compete with bG17 for the binding site, and the observation that bG17, bGRP, and bNC did not bind to other positively charged sites (e.g., collagen, red blood corpuscles), demonstrated that binding was not due to nonspecific electrostatic interactions alone. Binding of bG17 to a CCK(B)/gastrin-type receptor was ruled out when specific receptor antagonists failed to block binding. Conclusions: The particulate nature of the binding site suggests a secretory substance. If so, eosinophils might use that substance to destroy, neutralize, or control the activity of peptide hormones bound to it in the extracellular space.",
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