Factors restricting maximal preservation of neuronal glycogen after perfusion fixation with dimethyl sulfoxide and iodoacetic acid in Bouin's solution. Histochemical observations in the brain of the Netherlands dwarf rabbit.
Thirty seconds after an initial intracardial epinephrine injection, deeply anesthetized animals are perfused consecutively with saline, Bouin's and 100% ethanol solutions, each containing 1% or 5% DMSO (Me2SO) and 0.01 M iodoacetic acid. In the Netherlands dwarf rabbit and the guinea pig, a maximal preservation of dimedone PAS-stainable, saliva-digestible glycogen is achieved, without signs of polarization of glycogen, in many neuronal and neuroglial cells occupying either brain stem nuclei or occasionally narrow perivascular zones. Tentatively, these results are ascribed to a combined effect of (a) the alleged capacity of DMSO to accelerate fixation and to suppress activation of adenylate cyclase and (b) the rapid action of Bouin's solution so that the glycogen particles become instantaneously enclosed in situ in a skeleton of coagulated proteinaceous elements. The paradoxical over-all reduction in preservation of neuronal and astrocytic glycogen may be associated either with a demonstrable loss of the fixative into the peripheral vasculature, because of contrary actions of DMSO and epinephrine, or with a transvascular passage of epinephrine resulting in neuronal glycogenolysis where the blood-brain barrier is absent or affected by DMSO. Other defects are the occurrence of myriad pericapillary foci of inadequate tissue preservation, rare petechial hemorrhages, post mortem fat emboli, and ubiquitous Buscaino plaques. Despite these adverse results preventing utilization of this technique in systematic histochemical investigations on neuronal glycogen, remarkable qualitative characteristics such as the neurons' capacity to store glycogen throughout their perikarya have been revealed.
D000085:Acetates; D019342:Acetic Acid; D000818:Animals; D001921:Brain / Q000166:cytology; D001923:Brain Chemistry; D002415:Cats; D003593:Cytoplasm / Q000166:cytology; D004121:Dimethyl Sulfoxide; D005404:Fixatives; D005557:Formaldehyde; D006003:Glycogen / Q000032:analysis*; D006168:Guinea Pigs; D006651:Histocytochemistry; D007461:Iodoacetates; D019807:Iodoacetic Acid; D009474:Neurons / Q000032:analysis*; D010477:Perfusion; D010851:Picrates; D011817:Rabbits; D013045:Species Specificity; D013194:Staining and Labeling
D000085:Acetates; D005404:Fixatives; D007461:Iodoacetates; D010851:Picrates; D005557:Formaldehyde; C026239:Bouin's solution; D006003:Glycogen; D019342:Acetic Acid; D019807:Iodoacetic Acid; D004121:Dimethyl Sulfoxide