P. Ninfali et al., Glucose-6-phosphate dehydrogenase in small intestine of rabbit: biochemical properties and subcellular localization, ACT HISTOCH, 103(3), 2001, pp. 287-303
Biochemical properties and cellular and subcellular distribution patterns o
f glucose-6-phosphate dehydrogenase (G6PD) were investigated in small intes
tine of rabbits. The specific activity of G6PD in fresh homogenates of smal
l intestine was 19 +/- 9 IU/g protein. This value did not change significan
tly after dialysis. The kinetic and electrophoretic properties of the parti
ally purified enzyme were similar to those found in other rabbit tissues. E
nzyme histochemical. analysis of G6PD activity using the tetrazolium salt m
ethod showed high activity in epithelial cells of villi and crypts of Liebe
rkuhn. The activity in acinar cells of Brunner's glands was lower than that
in epithelium, whereas cells of the muscularis externa showed a very low a
ctivity. Immunohistochemical analysis showed that the amounts of G6PD prote
in were lower in the epithelium than in Brunner's glands and muscularis ext
erna. The differences between distribution patterns of activity and protein
of G6PD may reflect the presence of inactive enzyme molecules in Brunner's
glands and muscularis externa or posttranslational activation of G6PD in e
pithelium. Electron microscopic immunocytochemical analysis performed with
gold-labelled antibodies showed the presence of G6PD protein throughout the
cytoplasm and at smooth endoplasmic reticulum in enterocytes. In Paneth ce
lls and cells of Brunner's glands, G6PD was found in the cytoplasm, at roug
h endoplasmic reticulum and Golgi complex. Immunolabelling was not found in
mitochondria or nuclei. Our findings show that G6PD is heterogeneously dis
tributed in cells of the small intestine and that the enzyme is associated
with rough and smooth endoplasmic reticulum to support synthetic functions
in these compartments by NADPH production.