Regional distribution of soluble calcium activated proteinase activity in neonatal and adult rat brain.
Calcium dependent proteolytic activity in the soluble fraction of various rat brain regions was assayed using 14C-radiolabelled denatured casein as a substrate. Two forms of activity, distinguishable by their calcium requirement for half maximal activation (5 and 80 microM), were found; both were blocked by sulfhydryl alkylating agents and thiol proteinase inhibitors. Preincubation at 58 degrees C for 10 min also eliminated the high threshold activity. These characteristics are identical to those reported for calcium activated neutral proteinases ('calpains') found in other tissues. Calpain activity varied markedly across brain regions. The greatest values for the high threshold enzyme were found in pons-medulla followed by cerebellum/mesencephalon and finally the telencephalon. The low threshold enzyme had low levels of activity throughout the brainstem and diencephalon and was barely detectable in telencephalic structures. In contrast, a previously described endogenous inhibitor of calpain, 'calpastatin', was found not to vary in its activity across brain regions. Calpain activity was high in the prenatal brain, but while the hindbrain maintained high levels of activity into adulthood, the activity in the forebrain dropped 80% during the early postnatal period. The differences between forebrain and hindbrain activity levels were evident during the first 5 days of the neonatal period, suggesting that glial cell differentiation is not responsible for the regional variations found in the adult. These results are discussed with regard to the possibility that the turnover of anatomical structures differs between brain regions.
Simonson
L
L
Baudry
M
M
Siman
R
R
Lynch
G
G
D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism*; D001921:Brain / Q000201:enzymology*; D002154:Calpain; D010450:Endopeptidases / Q000378:metabolism*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility; D014018:Tissue Distribution
D010450:Endopeptidases; D002154:Calpain