共查询到20条相似文献,搜索用时 31 毫秒
1.
Bischoff HA Borchers M Gudat F Duermueller U Theiler R Stähelin HB Dick W 《The Histochemical journal》2001,33(1):19-24
Growing evidence suggests that intracellular vitamin D receptors are present in skeletal muscle tissue mediating vitamin D hormone response. The aim of the work reported here was to investigate the in situ expression of 1,25-dihydroxy vitamin D3 receptor in human skeletal muscle tissue. Intraoperative periarticular muscle biopsies were taken from 20 female orthopaedic patients (17 middle-aged and elderly patients receiving total hip arthroplasty due to osteoarthritis of the hip or an osteoporotic hip fracture and 3 young patients who received back surgery). The immunohistological distribution of the vitamin D3 receptor was investigated using a monoclonal rat antibody to the receptor (Clone Nr. 9A7). The receptor-positive nuclei were quantified by counting 500 nuclei per biopsy. Strong intranuclear immunostaining of the vitamin D receptor was detected in human muscle cells. Biopsies of hip patients had significantly fewer receptor-positive nuclei compared to those of back surgery patients (Mann–Whitney U-test: p = 0.0025). VDR expression (number of antigen-positive nuclei) was significantly correlated with age (coefficient of correlation = 0.46; p = 0.005), but not with 25-hydroxyvitamin D or 1,25-dihydroxyvitamin D levels. The data clearly demonstrate presence of nuclear 1,25-dihydroxyvitamin D3 receptor in human skeletal muscle. To our knowledge this is the first in situ detection of the receptor in human skeletal muscle. The difference in the expression of the receptor between hip and spinal muscle biopsies might be explained by age or location. Further research is needed in order to evaluate whether vitamin D3 receptor expression in human skeletal muscle is age-dependent and varies between different muscles. 相似文献
2.
H.A. Bischoff M. Borchers F. Gudat U. Duermueller R. Theiler H.B. Stähelin W. Dick 《Journal of molecular histology》2001,33(1):19-24
Growing evidence suggests that intracellular vitamin D receptors are present in skeletal muscle tissue mediating vitamin D hormone response. The aim of the work reported here was to investigate the in situ expression of 1,25-dihydroxy vitamin D3 receptor in human skeletal muscle tissue. Intraoperative periarticular muscle biopsies were taken from 20 female orthopaedic patients (17 middle-aged and elderly patients receiving total hip arthroplasty due to osteoarthritis of the hip or an osteoporotic hip fracture and 3 young patients who received back surgery). The immunohistological distribution of the vitamin D3 receptor was investigated using a monoclonal rat antibody to the receptor (Clone Nr. 9A7). The receptor-positive nuclei were quantified by counting 500 nuclei per biopsy. Strong intranuclear immunostaining of the vitamin D receptor was detected in human muscle cells. Biopsies of hip patients had significantly fewer receptor-positive nuclei compared to those of back surgery patients (Mann–Whitney U-test: p = 0.0025). VDR expression (number of antigen-positive nuclei) was significantly correlated with age (coefficient of correlation = 0.46; p = 0.005), but not with 25-hydroxyvitamin D or 1,25-dihydroxyvitamin D levels. The data clearly demonstrate presence of nuclear 1,25-dihydroxyvitamin D3 receptor in human skeletal muscle. To our knowledge this is the first in situ detection of the receptor in human skeletal muscle. The difference in the expression of the receptor between hip and spinal muscle biopsies might be explained by age or location. Further research is needed in order to evaluate whether vitamin D3 receptor expression in human skeletal muscle is age-dependent and varies between different muscles. 相似文献
3.
The binding of metabolites of vitamin D and their analogs to the 3.7S chick intestinal cytosol receptor protein has been specifically studied by competitive binding techniques and polyethylene glycol precipitation of the complex. The structural requirements for the interaction between the vitamin D molecule and the receptor could be assessed without the nuclear chromatin binding step. These measurements have shown that 1,25-dihydroxyvitamin D3 and 1,25-dihydroxyvitamin D2 are equally competitive and are the most active. Of the structural features of the compounds, the 1α-hydroxyl is most important followed by the 25-hydroxyl and the 3β-hydroxyl. The addition of a second hydroxyl near carbon 25 markedly reduces binding whether on the 26 carbon or the 24 carbon. A hydroxyl on C-24 could substitute to some degree for the 25-hydroxyl inasmuch as 24-hydroxyvitamin D3 was much more effective than vitamin D3 but less effective than 25-hydroxyvitamin D3. In general the patterns of binding affinities correlated well with the biological activity of the various analogs strongly supporting a physiological role for the 1,25-dihydroxyvitamin D3 binding protein. It also suggests that of the two-step receptor mechanism, the structural specificity is located in the initial interaction of the 1,25-dihydroxyvitamin D3 and the cytosol receptor. 相似文献
4.
M Zile E C Bunge L Barsness S Yamada H K Schnoes H F DeLuca 《Archives of biochemistry and biophysics》1978,186(1):15-24
Autoradiography of frozen sections of intestinal tissue taken from rachitic chickens given a single intravenous dose of 1,25-dihydroxy[23,243H]vitamin D3 (650 pmol, 78 Ci/ mmol) has been carried out. Specific localization of label in the nuclei of intestinal villi and crypt cells could be demonstrated at 2.5 to 6 h postinjection. In contrast, no concentration or localization of radioactivity could be detected in intestinal muscle, liver, and skeletal muscle. These results strongly support the concept that the function of 1,25-dihydroxyvitamin D3 in intestine is mediated by a nuclear mechanism. 相似文献
5.
R U Simpson R Weishaar 《BioEssays : news and reviews in molecular, cellular and developmental biology》1986,4(2):65-70
Recent studies have yielded new insights into the critical importance of adequate vitamin D3 intake and metabolism. Investigations of the actions of 1,25-dihydroxyvitamin D3 (calcitriol) on novel target tissues has revealed that this hormone has functions other than its recognized action in regulating blood calcium and phosphate levels. Reports have characterized calcitriol receptors and activities in organs and tissues as diverse as pancreas, skeletal and heart muscle, blood cells, brain, skin, pituitary, parathyroid, kidney, bone and intestine. These studies suggest functions for calcitriol as varied as the regulation of insulin and prolactin secretion, muscle contractility, immune cell metabolism, melanin synthesis and differentiation of blood cells. This information may ultimately help us to understand the etiologies of several kinds of organ dysfunction and lead to the development of tissue-specific agents for new therapies. 相似文献
6.
7.
Summary Rats and mice fed a vitamin D-deficient or vitamin D-complete diet were injected with 3H 1,25 (OH)2 vitamin D3. Autoradiograms prepared from cross sections through the neck region revealed nuclear concentration of radioactivity strongest in parathyroid chief cells, occasionally in thyroid follicular epithelial and interfollicular cells, in the epithelium of tubular remnants of the ultimobranchial body, in epithelium of the esophagus, in chondrocytes of tracheal cartilage, and in myoepithelial cells of tracheal glands. In the thyroid, most of the follicle epithelial cells did not show nuclear concentration of radioactivity which occurred only occasionally and predominantly in follicles located in marginal positions. Thyroglobulin in lumina of thyroid follicles contained varying amounts of radioactivity that correspond to the diameter of the follicles, with relatively high amounts in large follicles and little or no radioactivity in small follicles. Competition with excess of unlabeled 1,25 (OH)2 vitamin D3 abolished nuclear radioactivity, but not the radioactivity in the colloid, while 25 (OH) vitamin D3 did not affect either. When a combination of autoradiography and immunohistochemistry was applied, follicular and parafollicular C-cells positive for calcitonin antibodies, did not show nuclear concentration of radioactivity. Tubular remnants of ultimobranchial bodies, however, showed distinct nuclear labeling, but did not stain, or only weakly stain, with antibodies to calcitonin. When 3H 25 (OH) vitamin D3 was injected, no nuclear concentration of radioactivity was noted in any of the tissues.The results from these histochemical studies suggest the existence of nuclear receptors and direct genomic effects of 1,25 (OH)2 vitamin D3 in heterogeneous tissues of the neck region, which include parathyroid chief cells, myoepithelial cells of tracheal glands, chondrocytes of tracheal cartilage, epithelial cells of esophagus, and certain thyroid follicle epithelial cells. No evidence could be obtained for nuclear receptors in C-cells and cells of striated muscle. 相似文献
8.
In agreement with previous reports, chick intestinal calcium-binding protein does not appear in the chick embryo until 1 day after hatching while intestinal alkaline phosphatase begins to appear at 19–20 days of embryonic life. The ability of chick embryo to metabolize vitamin D3 to 25-hydroxyvitamin D3, 1,25-dihydroxyvitamin D3, and 24,25-dihydroxyvitamin D3 is present at least by day 18 of embryonic life as demonstrated by in vivo and in vitro techniques. It also illustrates that metabolism of vitamin D3 was not the limiting factor in the appearance of calcium-binding protein and alkaline phosphatase in intestine. Instead, the uptake of 1,25-dihydroxyvitamin D3 by the duodenum was very low prior to hatching, even though significant amounts were present in the yolk sac. Injection of a physiological dose of 1,25-dihydroxyvitamin D3 to chick embryo at 9 days failed to stimulate appearance of calcium binding protein by 18 days of embryonic life. Thus, it appears that either the normal mechanism for transport of 1,25-dihydroxyvitamin D3 to intestine or its receptors in intestine may not be present prior to day 18–19.A large fraction of radioactive vitamin D3 injected into the yolk sac was found esterified especially in the embryonic liver. The significance of this is not yet understood.Injection of 1,25-dihydroxyvitamin D3 at 325 pmoles/per egg at 9 days resulted in 70% mortality of embryos while a 32-pmole dose resulted in no significant increase in mortality. The basis for this toxicity is not yet understood. 相似文献
9.
Pyloric gastrin-producing cells and pyloric sphincter muscle cells are nuclear targets for 3H 1,25(OH)2 vitamin D3 总被引:1,自引:0,他引:1
Summary Autoradiographic studies were conducted to identify and characterize target cells for 1,25(OH)2 vitamin D3 in the pyloric region of rats and mice. After injection of 3H 1,25(OH)2 vitamin D3, nuclear concentration of radioactivity was observed in nuclei of duodenal epithelium and certain cells of pyloric glands, while most of the epithelial cells in the pyloric and gastric glands did not show nuclear labeling. In combined immunohistochemical studies, cells in the pyloric glands that showed nuclear concentration of radioactivity, were stained in their cytoplasm with antibodies to gastrin. Also, cells of the pyloric sphincter muscle showed nuclear labeling, in contrast to cells of the duodenal muscularis, which remained unlabeled under the conditions of the experiments. The results indicate that the cells with nuclear radioactivity contain receptors for 1,25(OH)2 vitamin D3 and suggest that gastrin secretion and pyloric muscle functions are regulated by a direct action of 1,25(OH)2 vitamin D3 on these cells. 相似文献
10.
Barbara E. Kream Robert D. Reynolds Joyce C. Knutson John A. Eisman Hector F. DeLuca 《Archives of biochemistry and biophysics》1976,176(2):779-787
The binding of 25-hydroxy-[26,27-3H]vitamin D3 and 1,25-dihydroxy-[26,27-3H]vitamin D3 to the cytosol of intestinal mucosa of chicks and rats has been studied by sucrose gradient analysis. The cytosol from chick mucosa showed variable binding of 1,25-dihydroxyvitamin D3 to a 3.0S macromolecule which has high affinity and low capacity for this metabolite. However, when the mucosa was washed extensively before homogenization, a 3.7S macromolecule was consistently observed which showed considerable specificity and affinity for 1,25-dihydroxyvitamin D3. Although 3.7S binders for 1,25-dihydroxyvitamin D3 could also be located in other organs, competition experiments with excess nonradioactive 1,25-dihydroxyvitamin D3 suggested that they were not identical to the 3.7S macromolecule from intestinal mucosal cytosol. As the 3.7S macromolecule was allowed to stand at 4 °C with bound 1,25-dihydroxy-[3H]vitamin D3, the 1,25-dihydroxy-[3H]vitamin D3 became increasingly resistant to displacement by non-radioactive 1,25-dihydroxyvitamin D3. The 1,25-dihydroxy-[3H]vitamin D3 remained unchanged and easily extractable with lipid solvents through this change, making unlikely the establishment of a covalent bond. Unlike the chick, mucosa from rats yielded cytosol in which no specific binding of 1,25-dihydroxy-[3H]vitamin D3 was detected. Instead, a 5-6S macromolecule which binds both 1,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3 was found. This protein which was also found in chick mucosa shows preferential binding for 25-hydroxyvitamin D3. It could be removed by washing the mucosa with buffer prior to homogenization which suggests that it may not be a cytosolic protein. Although the 3.7S protein from chick mucosa has properties consistent with its possible role as a receptor, the 5-6S macromolecule does not appear to have “receptor”-like properties. 相似文献
11.
Cytosol fractions prepared from rachitic chick kidney and pancreas were analyzed for binding of vitamin D3 metabolites by sucrose density gradient centrifugation. Both cytosol fractions were found to contain a 3.6S macromolecule which specifically binds 1,25-dihydroxy[3H] vitamin D3 and in addition a 5 to 6S macromolecule which binds 25-hydroxy[3H]vitamin D3. Sucrose gradient analysis of a KCl extract prepared from kidney or pancreas chromatin resulted in a peak (3.6S) of bound 1,25-dihydroxyvitamin D3 which could not be distinguished from the cytoplasmic binding component. The interaction of 1,25-dihydroxy[3H]vitamin D3 with the cytoplasmic binding component of both tissues occurred at low concentrations of hormone with high affinity. 相似文献
12.
M R Walters 《Biochemical and biophysical research communications》1981,103(2):721-726
Sucrose density gradient analysis was utilized to determine whether 1,25-dihydroxyvitamin D3 receptors are present in the rat uterus. A distinct 3.6S [3H]1,25-dihydroxyvitamin D3 binding component was observed in chromatin extracts of estrogen-primed, ovariectomized rat uteri. Binding to this putative 1,25-dihydroxyvitamin D3 receptor was inhibited by excess 1,25-dihydroxyvitamin D3, but not by 25-hydroxyvitamin D3, estradiol-17β, promegestone, or cortisol. Low levels of the receptor seemed to be present in the unprimed uterus. Estrogen injection significantly increased the number of 1,25-dihydroxyvitamin D3 receptors and progesterone co-administration reduced, but did not abolish, this effect. 相似文献
13.
Cytosol prepared from small intestine of vitamin D-sufficient rabbits contains a specific high-affinity binding protein for 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). This binding protein sediments at 3.0–3.5 S in sucrose density gradients containing 0.3 m KCl. Scatchard analysis using intestinal cytosol demonstrated a Kd of 0.05 nm and a maximum binding capacity of 92 fmol/mg cytosol protein for 1,25(OH)2D3 at 4°C. Competitive binding studies with various metabolites of vitamin D showed a relative binding affinity of this protein for 1,25(OH)2D3 > 25-hydroxy-vitamin D3 > vitamin D3. With 200 μg of rabbit intestinal cytosol protein, as little as 1.0–2.5 pg of 1,25(OH)2D3 reproducibly displaced the tracer sterol from the binding protein. Analyses of human plasma 1,25(OH)2D3 content yielded values consistent with published results. The vitamin D-replete rabbit provides a convenient, plentiful, and inexpensive source of binding protein for 1,25(OH)2D3 assays. 相似文献
14.
Helen L. Henry 《Biochemical and biophysical research communications》1977,74(2):768-774
The primary culture of kidney cells from vitamin D deficient chicks is described. After four days in culture the cells reach confluency and retain their ability to metabolize 25-hydroxyvitamin D3 to 1,25-dihydroxyvitamin D3. Addition of one unit of bovine parathyroid hormone to the culture medium for 48 hours prior to assay had no effect on the cells' ability to produce 1,25-dihydroxy vitamin D3, whereas after 24 hours in the presence of 5×10?8 1,25-dihydroxyvitamin D3 the cells produced not this metabolite, but 24,25-dihydroxyvitamin D3. This cell culture system will allow the investigation of the regulation of renal 25-hydroxyvitamin D3 metabolism under controlled conditions. 相似文献
15.
Dr. Walter E. Stumpf Sam A. Clark Madhabananda Sar Hector F. DcLuca 《Cell and tissue research》1984,238(3):489-496
Summary Tritium-labeled 1,25 (OH2) vitamin D3, when injected into vitamin D-deficient adult and pregnant rats is concentrated and retained strongest in nuclei of cells in the outer root sheath of the hair, followed by the stratum granulosum, spinosum, and basale of the epidermis. In the hair follicle, in addition to the most heavily labeled outer root sheath, nuclear labeling exists also in cells of the hair bulb and of the inner root sheath, as well as in basal cells of the sebaceous gland. In contrast, cells of the dermal papilla and the connective tissue of the dermis are generally unlabeled, except for labeled cells in the outer connective tissue sheath at the infundibulum of vibrissae of 20-day fetal rats and a few scattered labeled cells in the dermis, probably macrophages. In the developing hair, in 18- and 20-day fetal rats, a distinct topographic pattern of labeled cells can be seen, which is characteristic of the different stages of hair follicle development. In the hair germ, heavily labeled cells appear first in the stratum spinosum. In the hair peg, they remain in this position in its juxtaepidermal portion; however, when a dermal papilla develops, heavily labeled cells assume a marginal position. This suggests a sequential epidermal-epidermal and mesenchymal-epidermal receptor induction. Injection of tritium labeled 25 (OH) vitamin D3 did not show nuclear concentration in these tissues and excess unlabeled 25 (OH) vitamin D3 — unlike excess 1,25 (OH2) vitamin D3 — did not prevent nuclear uptake of tritium labeled 1,25 (OH2) vitamin D3. The results indicate differential effects of 1,25 (OH2) vitamin D3 on different structures in the epidermis and dermis.Supported by US PHS grant PCM8200569 相似文献
16.
Specific binding proteins for 1,25-dihydroxyvitamin D3 were identified in bovine mammary tissue obtained from lactating and non-lactating mammary glands by sucrose density gradient centrifugation. The macromolecules had characteristic sedimentation coefficients of 3.5-3.7 S. The interaction of l,25-dihydroxy[3H]vitamin D3 with the macromolecule of the mammary gland cytosol occurred at low concentrations, was saturable, and was a high affinity interaction (Kd = 4.2 × 10?10M at 25 °C). Binding was reversed by excess unlabeled 1,25-dihydroxyvitamin D3, was destroyed by heat and/or incubation with trypsin. It is thus inferred that this macromolecule is protein as it is not destroyed by ribonuclease or deoxyribonuclease. 25-hydroxyvitamin D3, 24,25-dihydroxyvitamin D3, and vitamin D3 did not effectively compete with 1,25-dihydroxyvitamin D3 for binding to cytosol of mammary tissue at near physiological concentrations of these analogs, thus demonstrating the specificity of the binding protein for 1,25-dihydroxyvitamin D3. In vitro subcellular distribution of 1,25-dihydroxy[3H]vitamin D3 demonstrated a time- and temperature-dependent movement of the hormone from the cytoplasm to the nucleus. By 90 min at 25 °C 72% of the 1,25-dihydroxy[3H]vitamin D3 was associated with the nucleus. In addition a 5–6 S macromolecule which binds 25-hydroxy[3H]vitamin D3 was demonstrated in mammary tissue. Finally, it is possible that the receptor-hormone complex present in mammary tissue may function in a manner analogous to intestinal tissue, resulting in the control of calcium transport by 1,25-dihydroxyvitamin D3 in this tissue. 相似文献
17.
《Bioscience, biotechnology, and biochemistry》2013,77(1):312-315
1α,25-dihydroxyvitamin D3 [1,25-(OH)2D3] phosphorylates the extracellular signal-regulated kinase (ERK), a member of the mitogen-activated protein kinase (MAPK) family, within 30 sec in primary cultured chick skeletal muscle cells. MAPK of HeLa cell lines, which had been stably transfected with a cDNA library derived from mRNA of chick skeletal muscle cells, was also rapidly phosphorylated by 1,25-(OH)2D3. These cell lines have the potential to be a good tool for further investigation of rapid non-genomic mechanism activated by 1,25-(OH)2D3. 相似文献
18.
Y. Tanaka J.K. Wichmann H.F. De Luca Y. Kobayashi N. Ikekawa 《Archives of biochemistry and biophysics》1983,225(2):649-655
To evaluate possible functional roles for 24,25-dihydroxyvitamin D3, 24,24-difluoro-25-hydroxyvitamin D3 has been synthesized and shown to be equally as active as 25-hydroxyvitamin D3 in all known functions of vitamin D. The use of the difluoro compound for this purpose is based on the assumption that the C-F bonds are stable in vivo and that the fluorine atom does not act as hydroxyl in biological systems. No 24,25-dihydroxyvitamin D3 was detected in the serum obtained from vitamin D-deficient rats that had been given 24,24-difluoro-25-hydroxyvitamin D3, while large amounts were found when 25-hydroxyvitamin D3 was given. Incubation of the 24,24-difluoro compound with kidney homogenate prepared from vitamin D-replete chickens failed to produce 24,25-dihydroxyvitamin D3, while the same preparations produced large amounts of 24,25-dihydroxyvitamin D3 from 25-hydroxyvitamin D3. Kidney homogenate prepared from vitamin D-deficient chickens produced 24,24-difluoro-1,25-dihydroxyvitamin D3 from 24,24-difluoro-25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 from 25-hydroxyvitamin D3. In binding to the plasma transport protein for vitamin D compounds, 24,24-difluoro-25-hydroxyvitamin D3 is less active than 25-hydroxyvitamin D3 and 24R,25-dihydroxyvitamin D3. In binding to the chick intestinal cytosol receptor, 24,24-difluoro-25-hydroxyvitamin D3 is more active than 25-hydroxyvitamin D3 which is itself more active than 24R,25-dihydroxyvitamin D3. The 24,24-difluoro-1,25-dihydroxyvitamin D3 is equal to 1,25-dihydroxyvitamin D3, and both are 10 times more active than 1,24R,25-trihydroxyvitamin D3 in this system. These results provide strong evidence that the C-24 carbon of 24,24-difluoro-25-hydroxyvitamin D3 cannot be hydroxylated in vivo, and, further, the 24-F substitution acts similar to H and not to OH in discriminating binding systems for vitamin D compounds. 相似文献
19.
Dr. Samuel A. Clark Walter E. Stumpf Charles W. Bishop Hector F. DeLuca Dong H. Park Tong H. Joh 《Cell and tissue research》1986,243(2):299-302
Summary Target cells for 1,25-dihydroxyvitamin D3 were demonstrated in the adrenal medulla by frozen-section autoradiography. The appearance of these target cells was age-dependent in neonatal mice. Immunocytochemical staining for phenylethanolamine-N-methyltransferase revealed that both epinephrine and non-epinephrine cells concentrate 1,25-dihydroxyvitamin D3 in their nuclei. In contrast, immunocytochemical staining for vitamin D-dependent calcium-binding protein (D-CaBP) demonstrated that D-CaBP immunoreactivity is localized in only a small percentage of adrenomedullary cells, in mice and rats. Comparison of PNMT and D-CaBP immunoreactivities in sequential sections showed that epinephrine-producing cells do not contain D-CaBP. These results indicate that adrenal medullary cells have receptors for 1,25-dihydroxyvitamin D3 and that 1,25-dihydroxyvitamin D3 may directly affect certain functions of these endocrine cells.Supported by US PHS grant PCM8200569 and postdoctoral training grant AM07240-02 相似文献
20.
Robert U. Simpson Joseph K. Wichmann Herbert E. Paaren Heinrich K. Schnoes Hector F. DeLuca 《Archives of biochemistry and biophysics》1984,230(1):21-29
A primary confluent culture of epithelial cells from rat kidney has been developed. These cells possess a 3.2–3.4 S high-affinity, low-capacity binding protein for 1,25-dihydroxyvitamin D3. They metabolize 25-hydroxyvitamin D3 to at least five metabolites. Two have been identified as 1,25-dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3. Two others have been identified by means of physical data and cochromatography as trans 19-nor-10-oxo-25-hydroxyvitamin D3 and the other as its cis isomer. These two “metabolites” have not been observed in vivo, but one of them (cis) comigrates with 1,25-dihydroxyvitamin D3 on straight-phase high-performance liquid chromatography. Thus, mere cochromatography on high-performance liquid chromatography is not sufficient to identify critical vitamin D metabolites. 相似文献