Histochemical localization of neurons with zinc-permeable AMPA/kainate channels in rat brain slices.
Zinc modulates neurotransmission and may trigger neurodegeneration following brain injuries. Therefore, it is important to understand zinc dynamics in an anatomical context. Using a histochemical procedure on stimulated slices, we visualized zinc influx into neurons 'in situ'. Hippocampal, neocortical and cerebellar slices were loaded with zinc and stimulated with compounds known to open zinc-permeable channels. Zinc was then visualized by histochemical precipitation methods. Kainate stimulation labelled CA1 hippocampal pyramidal neurons, as well as subpopulations of interneurons in the hilus, CA1 and CA3 fields. Interneurons in the neocortex and many cell types of the cerebellum were also labelled. However, neither NMDA nor 50 mM K(+) stimulation resulted in comparable zinc accumulation in neurons. Immunofluorescent colocalization of parvalbumin with kainate-induced zinc staining in the hippocampus and neocortex showed a subset of zinc-sensitive neurons, positive for parvalbumin. These results confirm that zinc permeation through calcium-permeable AMPA/kainate channels constitutes a predominant route of zinc entry into different cell types. Furthermore, this technique provides a useful and versatile histochemical approach to assess the cell subpopulations of the central nervous system particularly sensitive to zinc accumulation under normal or pathological conditions.
Martínez-Galán
Juan Ramón
JR
Facultad de Medicina and Centro Regional de Investigación Biomédica, Universidad de Castilla-La Mancha, Edificio Benjamín Palencia, Campus de Albacete, 02071, Albacete, Spain.
Díaz
Carmen
C
Juiz
José M
JM
D000818:Animals; D001921:Brain / Q000033:anatomy & histology*; D001923:Brain Chemistry / Q000187:drug effects*; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D019579:Neocortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D018091:Receptors, AMPA / Q000378:metabolism*; D018092:Kainic Acid Receptors / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism*
D018091:Receptors, AMPA; D018092:Kainic Acid Receptors; D015032:Zinc