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Histochemistry

, Volume 74, Issue 4, pp 457–467 | Cite as

Enzymatic cleavage prior to antibody incubation as a method for neuropeptide immunocytochemistry

  • R. Martin
  • M. Schäfer
  • K. H. Voigt
Article

Summary

When deplasticized Epon sections were treated with endo- and/or exopeptidases prior to incubation with antibodies, the neuropeptide immunoreactivity of secretory nerves was often altered in a predictable way. Cleavage of neurosecretory material in octopus nerves by trypsin and carboxypeptidase-B enhanced enkephalin-like immunoreactivity, while Molluscan neuropeptide-like immunoreactivity was prevented by tryptic cleavage. The enzyme effects indicated the occurrence of a heptapeptide (Tyr-Gly-Gly-Phe-Met/Leu-Arg-Phe) that contains both the enkephalin and the Molluscan neuropeptide sequence. Vasopressin terminals of the rat neurohypophysis, which presumably contain enkephalin precursor sequences, exhibited enkephalin-like immunostaining after tryptic cleavage. ACTH/β-endorphin cells of the rat intermediate pituitary, which synthesize the enkephalin sequence at the N-terminus of β-endorphin, exhibited enkephalin-like immunoreactivity when sections were treated with α-chymotrypsin or trypsin, but not after incubation with leucine-aminopeptidase or carboxypeptidase-B. Enkephalin-like immunostaining could not be induced in any way in ACTH/β-endorphin cells of the anterior pituitary. Enzymatic cleavage may give additional information in immunocytochemical localization studies on neuropeptide sequences in secretory nerves and hormonal granules.

Keywords

Vasopressin Anterior Pituitary Enzymatic Cleavage Immunocytochemical Localization Precursor Sequence 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

ACTH

adrenocorticotropic hormon

Ap

leucine-aminopeptidase

Ch

α-chymotrypsin

Cp

carboxypeptidase-B

enk

enkephalin (Tyr-Gly-Gly-Phe-Met/Leu)

Mnp

Molluscan neuropeptide (Phe-Met-Arg-PheNH2)

PBS

phosphate buffered saline

T

trypsin

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Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • R. Martin
    • 1
  • M. Schäfer
    • 1
  • K. H. Voigt
    • 1
  1. 1.Sektion Elektronenmikroskopie und Labor für NeuroendokrinologieUniversität UlmUlmFederal Republic of Germany

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