Article
Immunogold Staining Procedure for the Localisation of Regulatory Peptides
Autor
Varndell, I.M.
Tapia, Félix J.
Probert, L.
Buchan, A. M. J.
Gu, Jiang
De Mey, J.
Bloom, S. R.
Polak, J. M.
Institución
Resumen
The use of protein A- and IgG-conjugated colloidal gold staining methods for the immuno-localisation of peptide hormones and neurotransmitters at light - and electron microscope level are described and discussed. Bright-field and dark-ground illumination modes have been used to visualise the gold-labelled antigenic sites at the light microscope level. Immunogold staining procedures at the ultrastructural level using region-specific antisera have been adopted to localise specific molecular forms of peptides including gastrin (G17 and G34), glucagon and pro-glucagon, insulin and pro-insulin, in normal tissue and in tumours of the gastroenteropancreatic system. Similar methods have been used to demonstrate the heterogeneity of p-type nerves in the enteric nervous system. Vasoactive intestinal polypeptide (VIP) has been localised to granular sites (mean±S.D. granule diameter=98 ± 19 nm) in nerve terminals of the enteric plexuses and in tumour cells of diarrhoeogenic VIP•producing neoplasias (mean ±S.D granule diameter=126 ± 37 nm) using immunogold procedures applied to ultraviolet-cured ultrathin sections. Co-localisation of amines and peptides in carotid body type I cells and in chromaffin cells of normal adrenal medulla and phaeochromocytomas has also been demonstrated. Advantages of the immunogold procedures over alternative immunocytochemical techniques are discussed.