NADPH-consuming enzymes correlate with glucose-6-phosphate dehydrogenase in Purkinje cells: an immunohistochemical and enzyme histochemical study of the rat cerebellar cortex.
In cerebellum of the adult rat, glucose-6-phosphate dehydrogenase (G6PD) activity is particularly localized in Purkinje cells, showing lower activity in the molecular and granule cell layers. G6PD is the first and rate-limiting step of the hexose monophosphate shunt (HMS), which has the physiological role of providing NADPH for reductive biosynthesis and detoxifying reactions. In this study, we searched for a possible correlation between G6PD and other NADPH-consuming enzymes, such as NADPH-cytochrome P450 reductase (P450R), glutathione reductase (GR) and NADPH-diaphorase (NADPH-d). This study was performed by means of immunohistochemistry and enzyme histochemistry followed by quantitative densitometric and confocal laser scanning microscopic analyses. Our results demonstrated that G6PD, P450R and GR have a similar distribution pattern characterized by the highest concentration of these enzymes in the somata of Purkinje cells, and by lower concentrations in the molecular and the granule cell layers. Moreover, in Purkinje cells, G6PD colocalized with both P450R and GR. NADPH-d activity showed a different distribution pattern when compared to the other enzymes, revealing the highest activity in the molecular layer and the lowest in Purkinje cells. Our results suggest a coordinated regulative mechanism of G6PD, P450R and GR based on the request of NADPH or on specific transcription factors.
Ferri
Paola
P
Institute of Morphological Sciences, University of Urbino Carlo Bo, I-61029 Urbino, Italy.
Biagiotti
Enrica
E
Ambrogini
Patrizia
P
Santi
Spartaco
S
Del Grande
Paolo
P
Ninfali
Paolino
P
D000704:Analysis of Variance; D000818:Animals; D002525:Cerebellar Cortex / Q000166:cytology* / Q000201:enzymology; D003720:Densitometry / Q000379:methods; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism*; D005980:Glutathione Reductase / Q000378:metabolism*; D006651:Histocytochemistry / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D018613:Microscopy, Confocal / Q000379:methods; D009249:NADP / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism*; D011689:Purkinje Cells / Q000201:enzymology* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D005904:Glial Fibrillary Acidic Protein; D009249:NADP; D005954:Glucosephosphate Dehydrogenase; D009251:NADPH-Ferrihemoprotein Reductase; D005980:Glutathione Reductase