Different localizations of Met-enkephalin-like immunoreactivity in rat forebrain and spinal cord using hydrogen peroxide and Triton X-100. Light microscopic study.
The histological distribution of met-enkephalin-like immunoreactivity was studied in the forebrain (particularly the striatum) and the spinal cord of the rat using the indirect peroxidase-labelled antibody method. In most experiments, vibratome sections of formaldehyde-fixed tissues and purified antibodies were used. The search for optimal conditions for the immunohistochemical reaction lead us to establish that met-enkephalin-containing perikarya of both untreated and colchicinized animals were better demonstrated when tissue were pre-treated with diluted hydrogen peroxide only. The additional treatment of these sections with Triton X-100 (or some other detergents) resulted in the near disappearance of the perikaryal immunoreactivity; on the contrary, numerous met-enkephalin containing nerve fibres and varicosities were then demonstrated in the same region. Using only the hydrogen peroxide treatment, we found numerous met-enkephalin-containing perikarya in the medial and ventral regions of the neostriatum. This distribution was prolonged caudally by the existence of a prominent group of stained somata in the ventral putamen-central nucleus of the amygdala. When intraventricular injections of colchicine were used, positive perikarya were more numerous within the striatum (the globus pallidus excepted) but their distribution was largely the same as in non injected animals. However, some new groups of somata were stained in this case in the forebrain (in the lateral septum, the olfactory tubercle and the hypothalamus particularly). In control animals only few met-enkephalin-containing perikarya were observed in the dorsal horn of the spinal cord when H2O2 pretreatment was used alone and they were numerous only when intraspinal injections of colchicine were performed. Met-enkephalin-containing fibres and varicosities, which were scattered in the whole neostriatum in the conditions used above, became very numerous when the tissue sections were incubated in the presence of Triton X-100. Their density increased markedly from the latero-dorsal to the medio-ventral regions but, in addition, an organization under the form of islands of stronger immunoreactivity was also evidenced. These islands were more numerous ventrally in the anterior neostriatum and in the central region of the "putamen." The dense plexus of immunoreactive nerve fibers forming "tube-like structures" which was always observed in the paleostriatum and in the cranial medial forebrain bundle (islands of Calleja) appeared more diffuse when detergents were used.(ABSTRACT TRUNCATED AT 400 WORDS)
D000818:Animals; D001921:Brain / Q000276:immunology*; D002421:Caudate Nucleus / Q000276:immunology; D003078:Colchicine; D003342:Corpus Striatum / Q000276:immunology; D004744:Enkephalin, Methionine / Q000276:immunology*; D006651:Histocytochemistry; D006861:Hydrogen Peroxide; D007124:Immunoenzyme Techniques; D017830:Octoxynol; D011092:Polyethylene Glycols; D051381:Rats; D011919:Rats, Inbred Strains; D013116:Spinal Cord / Q000276:immunology*; D014018:Tissue Distribution
D011092:Polyethylene Glycols; D004744:Enkephalin, Methionine; D017830:Octoxynol; D006861:Hydrogen Peroxide; D003078:Colchicine