Submitochondrial localization of kidney 25-hydroxycholecalciferol 1α-hydroxylase in vitamin D repleted weanling guinea pigs

1987 ◽  
Vol 65 (7) ◽  
pp. 673-676 ◽  
Author(s):  
Xue Yan ◽  
Guy Charette ◽  
Edgard E. Delvin

We have studied the submitochondrial localization of guinea-pig kidney 25-hydroxycholecalciferol 1α-hydroxylase. Treatment of the mitochondrial-enriched fraction with recrystallized digitonin produced mitoplasts bordered by a single membrane and with intact matrix. They contained nearly 90% of the 25-hydroxycholecalciferol 1α-hydroxylase activity and nearly 100% of the cytochrome-c: oxygen oxidoreductase. Amine: oxygen oxidoreductase activity remained mainly in the outer membrane fraction. These data show that 25-hydroxycholecalciferol 1α-hydroxylase has a distribution similar to that of steroid hydroxylases.

2000 ◽  
Vol 84 (4) ◽  
pp. 495-504 ◽  
Author(s):  
Katrien Rummens ◽  
Erik Van Herck ◽  
Rita van Bree ◽  
Roger Bouillon ◽  
F. André Van Assche ◽  
...  

Guinea-pig fetuses at term are mineralized to a degree comparable with human fetuses, which makes the guinea-pig an attractive animal model to study maternal–fetal interactions with regard to Ca and phosphate (P) homeostasis. We studied non-pregnant and pregnant (day 57) vitamin D-replete guinea-pigs, fed either a normal guinea-pig chow with 9·6 g Ca/kg and 4·9 g P/kg or a study diet with 2 g Ca/kg and 1 g P/kg (low-Ca–P diet) for 7–8 weeks. Both pregnancy and the low-Ca–P diet decreased plasma concentrations of 25-hydroxycholecalciferol (25(OH)D3), but increased total and free 1α,25-dihydroxycholecalciferol (1,25(OH)2D3), strongly suggesting an additive stimulation of 1α-hydroxylase activity. Maternal and fetal 25(OH)D3and 1,25(OH)2D3levels were highly correlated (r0·82 and 0·92 respectively, P<0·001). Dual-energy absorption X-ray absorptiometry (DXA) showed that both pregnancy and the low-Ca–P diet decreased bone mineral density (BMD) of the maternal femur, particularly at the distal metaphysis. Despite higher 1,25(OH)2D3concentrations and lower BMD, pregnant animals on the low-Ca–P diet were hypocalcaemic; blood Ca2+levels were inversely correlated with the number of fetuses in this group (r-0·93, P<0·001). Fetal growth as well as mineralization (assessed by whole-body and femoral DXA, bone histomorphometry and plasma–bone osteocalcin measurements) were unaltered in the low-Ca–P group. In conclusion, fetal mineralization proceeds normally but induces maternal hypocalcaemia in guinea-pigs with dietary restriction of Ca and P.


1978 ◽  
Vol 174 (1) ◽  
pp. 231-236 ◽  
Author(s):  
E Spanos ◽  
D I Barrett ◽  
K T Chong ◽  
I MacIntyre

Primary cultures of chick kidney cells convert 25-hydroxycholecalciferol into more-polar metabolites. Cells from vitamin D-deficient chicks have high 25-hydroxycholecalciferol 1 alpha-hydroxylase (1 alpha-hydroxylase) activity, but no 25-hydroxycholecalciferol 24-hydroxylase (24-hydroxylase) activity. Physiological concentrations of 1,25-dihydroxycholeclaciferol suppress 1 alpha-hydroxylase and induce 24-hydroxylase activity. The inhibition of 1 alpha-hydroxylase preceded the induction of 24-hydroxylase. In contrast, oestradiol-17 beta had no effect on the activity of either hydroxylase under a variety of experimental conditions. These results clearly demonstrate that 1,25-dihydroxycholecalciferol, but not oestrogen, acts directly on the kidney cells to regulate the metabolism of 25-hydroxycholecalciferol.


1980 ◽  
Vol 188 (3) ◽  
pp. 741-748 ◽  
Author(s):  
M Watford ◽  
P Vinay ◽  
G Lemieux ◽  
A Gougoux

The suppression by 3-mercaptopicolinate of gluconeogenesis from glutamine or 2-oxoglutarate in rat or dog kidney tubules did not affect the amount of these substrates undergoing complete oxidation. Furthermore, 3-mercaptopicolinate caused an accumulation of lactate in dog tubules. 3-Mercaptopicolinate abolished both gluconeogenesis and substrate oxidation in tubules from rabbit and guinea-pig kidney. These results imply the presence of an alternative pathway to phosphoenolpyruvate carboxykinase/pyruvate kinase for the production of pyruvate from citric-acid-cycle intermediates in the kidney cortex of rats and dogs but not in that of rabbits or guinea pigs. Oxaloacetate decarboxylase (present in the kidney cortex of all four species) or ‘malic’ enzyme (present in rat and dog but absent in rabbit and guinea-pig kidney cortex) could function in this role. Our observations indicate that ‘malic’ enzyme is probably implicated in this phenomenon. The lactate production observed in dog tubules in the presence of 3-mercaptopicolinate can be suppressed when aspartate formation is inhibited by 2-amino-4-methoxy-trans-but-3-enoic acid. This suggests that the provision of cytosolic NADH from citric-acid-cycle intermediates is facilitated by accumulation of aspartate acting as a ‘sink’ for cytosolic oxaloacetate.


1977 ◽  
Vol 38 (1) ◽  
pp. 47-54 ◽  
Author(s):  
R. Swaminathan ◽  
Barbara A. Sommerville ◽  
A. D. Care

1.As most of the studies on the regulation of renal 25-hydroxycholecalciferol-1-hydroxylase (25-HCC-1-hydroxylase) activity have been done in marginally-vitamin D-defieient animals and as it is known that vitamin D administration suppresses the specific activity of the 25-HCC-1-hydroxylase, it was decided to study the effect of dietary calcium on the activity of 25-HCC-1-hydroxylase and on Ca absorption in vitamin Dreplete chicks.2.Chicks, 10 d old, were given diets differing in their Ca contents (65 nmol cholecalciferol/kg diet) for 10 d and the activity of 25-HCC-1-hydroxylase in kidney homogenates, Ca absorption from the duodenum, Cabinding protein (CaBP) activity in the duodenal mucosa and plasma Ca and phosphate concentrations were all determined.3.The CaBP activity and the efficiency of Ca absorption both decreased with increasing dietary intake of Ca. Ca absorption and CaBP activity were significantly correlated (r 0.995, P < 0.01).4.The activity of 25-HCC-1-hydroxylase decreased as the dietary level of Ca increased and was significantly correlated with Ca absorption (r0.900, P < 005). The plasma Ca concentration and the activity of 25-HCC-1-hydroxylase were inversely related (r-0.940, P < 0.01).5.It is concluded that in the vitamin D-replete chick the efficiency of duodenal Ca absorption is regulated by the renal 25-HCC-1-hydroxylase activity via production of 1,25-dihydroxycholecalciferol and CaBP synthesis.


1975 ◽  
Vol 48 (3) ◽  
pp. 227-230
Author(s):  
I. M. A. Evans ◽  
K. W. Colston ◽  
L. Galante ◽  
I. MacIntyre

1. In vitamin D-deficient chicks both vitamin D3 and 1α-hydroxycholecalciferol markedly decrease renal 1-hydroxylase activity and induce 24-hydroxylase activity. 2. Actinomycin D abolishes both effects. 3. These results are consistent with feedback regulation of vitamin D3 metabolism by a direct nuclear action of the vitamin or its metabolites on the kidney cells.


1969 ◽  
Vol 112 (4) ◽  
pp. 407-413
Author(s):  
Morris Dillard ◽  
Philip K. Bondy

Radioactive androsterone was incubated with kidney slices from guinea pigs in a Ringer bicarbonate buffer. For analysis, the radioactivity was partitioned between chloroform and water for the separation of non-polar and polar steroids. After multiple chromatography procedures, the non-polar fraction was shown to contain androsterone and at least four other similar steroids, not identified in this study. The polar fraction yielded numerous conjugates from which androsterone glucuronide and androsterone sulphate in approximately equal quantities were isolated and characterized.


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