Immunocytochemical localization of glycogen phosphorylase kinase in rat brain sections and in glial and neuronal primary cultures.
The physiological function of brain glycogen and the role of phosphorylase kinase as a regulatory enzyme in the cascade of reactions associated with glycogenolysis in the brain have not been fully elucidated. As a first step toward elucidating such a function, we studied the localization of phosphorylase kinase in glial and neuronal primary cell cultures, and in adult rat brain slices, using a rabbit polyclonal antibody against skeletal muscle glycogen phosphorylase kinase. Immunocytochemical examination of rat astroglia-rich primary cultures revealed that a large number of cells were positive for glycogen phosphorylase kinase immunoreactivity. These cells were also positive for vimentin, a marker for immature glia, while they were negative for glial fibrillary acidic protein, a marker for mature astroglia, and for galactocerebroside, an oligodendroglial marker. Neurons in rat neuron-rich primary cultures did not show any kinase-positive staining. In paraformaldehyde-fixed adult rat brain sections, phosphorylase kinase immunoreactivity was detected in glial-like cells throughout the brain, with relatively high staining found in the cerebral cortex, the cerebellum, and the medulla oblongata. Phosphorylase kinase immunoreactivity could not be detected in neurons, with the exception of a group of large neurons in the brain stem, most likely belonging to the mesencephalic trigeminal nucleus. Phosphorylase kinase was also localized in the choroid plexus and to a lesser degree in the ependymal cells lining the ventricles. Phosphorylase kinase thus appears to have the same cellular distribution in nervous tissue as its substrates, i.e. glycogen phosphorylase and glycogen, which suggests that the physiological role of brain phosphorylase kinase is the mobilization of glycogen stores to fuel the increased metabolic demands of neurons and astrocytes.
Psarra
A M
AM
Institute of Biological Research and Biotechnology, the National Hellenic Research Foundation, 48 Vassileos Constantinou Avenue, 11635, Athens, Greece.
Pfeiffer
B
B
Giannakopoulou
M
M
Sotiroudis
T G
TG
Stylianopoulou
F
F
Hamprecht
B
B
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000201:enzymology*; D001921:Brain / Q000166:cytology* / Q000201:enzymology*; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques; D008297:Male; D009474:Neurons / Q000166:cytology / Q000201:enzymology*; D010764:Phosphorylase Kinase / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar
D010764:Phosphorylase Kinase