Binding of 1,25‐dihydroxy‐[<sup>3</sup>H]vitamin D<sub>3</sub> in nuclear and cytosol fractions of whole mouse skin in vivo and in vitro

Clemens, Thomas L.; Horiuchi, Noboru; Nguyen, Minh Van; Holick, Michael F.

doi:10.1016/0014-5793(81)80602-3

Cited by 25 publications

(5 citation statements)

References 20 publications

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“…This hormone acts on target tissues, such as the intestine, after binding to a specific cytosolic receptor protein. Recently, receptor-like binding proteins for 1,25-(OH) 2 D 3 have been demonstrated in intact mammalian skin (2)(3)(4)(5), raising the possibility that the hormone might function in this tissue. Subsequent studies by Eil and Marx (6) have shown that cultured human fibroblasts also contain a cytosolic 1,25-(OH) 2 D 3 binder and are capable of saturable nuclear uptake of hormone.…”

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confidence: 99%

Interaction of 1,25-Dihydroxyvitamin-D₃with Keratinocytes and Fibroblasts from Skin of Normal Subjects and a Subject with Vitamin-D-Dependent Rickets, Type II: A Model for Study of the Mode of Action of 1,25-Dihydroxyvitamin D₃*

Clemens

Adams

Horiuchi

et al. 1983

The Journal of Clinical Endocrinology & Metabolism

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194

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The binding of [ 3 H]l,25-dihydroxyvitamin D 3 (1,25-(OH) 2 -[ 3 H]D3) was examined in the cytosol of epidermal keratinocytes and dermal fibroblasts grown from normal human skin. Both cell types contained macromolecules with high affinity for 1,25-(OH) 2 D 3 , as demonstrated by sucrose density gradient centrifugation, saturation binding analysis, and DNAcellulose chromatography. Scatchard analysis of cytosol binding of the hormone yielded affinity constants of 1.0 x 10" 10 and 0.8 x 10" 10 M and binding capacities of 6.4 and 8.7 fmol/mg protein for fibroblasts and keratinocytes, respectively. In parallel studies, binding of 1,25-(OH) 2 -[ 3 H]D3 was evaluated in the cytosol from keratinocytes and fibroblasts cultured from a skin biopsy of a patient with vitamin D-dependent rickets, type II (DDR-II), an inheritable disorder characterized by extreme end-organ resistance to the action of 1,25-(OH)2D 3 . Multiple analyses by sucrose density gradient or saturation binding assays failed to reveal specific saturable binding of hormone, suggesting that in this disease the lack of an effective cytosolic receptor protein may account for end-organ insensitivity to the hormone. To evaluate the responsiveness of fibroblasts to 1,25-(OH) 2 D3, we studied the effect of 1,25-(OH) 2 D 3 on cell growth. 1,25-(OH) 2 D 3 (10~1 0 -10~6 M) caused a dose-dependent inhibition of cell growth in receptor-positive normal fibroblasts, whereas these doses of hormone did not affect the growth of receptor-defective DDR-II cells. Hence, the presence of an effective cytosolic receptor in normal skin fibroblasts may be necessary for expression of the growth inhibitory effect of 1,25-(OH) 2 D 3 . (J Clin Endocrinol Metab 56: 824, 1983) V ITAMIN D 3 is synthesized photochemically in the skin and then sequentially hydroxylated to 1,25-dihydroxyvitamin D 3 [1,25-(OH) 2 D 3 ], the biologically active form of the vitamin (1). This hormone acts on target tissues, such as the intestine, after binding to a specific cytosolic receptor protein. Recently, receptor-like binding proteins for 1,25-(OH) 2 D 3 have been demonstrated in intact mammalian skin (2-5), raising the possibility that the hormone might function in this tissue. Subsequent studies by Eil and Marx (6) have shown that

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confidence: 99%

Interaction of 1,25-Dihydroxyvitamin-D₃with Keratinocytes and Fibroblasts from Skin of Normal Subjects and a Subject with Vitamin-D-Dependent Rickets, Type II: A Model for Study of the Mode of Action of 1,25-Dihydroxyvitamin D₃*

Clemens

Adams

Horiuchi

et al. 1983

The Journal of Clinical Endocrinology & Metabolism

Self Cite

194

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“…Although vitamin D3 is synthesized by skin from endogenous 7-dehydrocholesterol (MacLaughlin et al, 1982), it is the resulting kidney metabolite, 1 a,25-dihydroxyvitamin D3 [la,25(OH)zD3] which is known to possess hormonal regulatory activities in various target organs via a receptor-mediated mechanism (DeLuca and Schnoes, 1976). Receptors for ~X ,~~( O H )~D~ have recently been reported in fresh and cultured human skin (Feldman et al, 19801, in cytosol prepared from rat (Simpson and DeLuca, 1980), and in nuclear and cytosol fractions of whole mouse skin in vivo and in vitro (Clemens et al, 1981), suggesting that this tissue, along with bone and intestine, is an additional target organ for this hormone. In fact, it was reported recently that the concentration of la,%(oH)zD3 binding macromolecule in mouse skin chromatin increased significantly during fetal and neonatal development (Horiuchi et al, 1985), thus prompting the suggestion that increase in this vitamin-chromatin binding may play a regulatory role in epidermal proliferation and differentiation during skin development.…”

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Novel regulatory actions of 1α,25‐dihydroxyvitamin D₃ on the metabolism of polyphosphoinositides in murine epidermal keratinocytes

Tang¹,

Ziboh²,

Isseroff³

et al. 1987

Journal Cellular Physiology

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The in vitro incubation of murine keratinocytes in the presence of 1 alpha,25-dihydroxyvitamin D3 enhanced the rapid hydrolysis of the prelabeled keratinocyte polyphosphoinositides (polyPtdIns) when compared to untreated cells. The rapid hydrolysis of the polyPtdIns and the release of the inositol phosphates (particularly InsP3 and InsP2) precede the onset of differentiation of these cells. These data therefore suggest that 1 alpha,25-dihydroxyvitamin D3 functions in vitro to initiate the rapid generation of InsP3 from cellular polyPtdIns; this in turn may mobilize intracellular Ca2+, thus providing the signal which program the murine keratinocytes from a proliferating mode into a differentiating mode.

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“…All had a normal karyotype, five were male and one was female. Hypertonic cytosol extracts were prepared by sonication followed by centrifugation at Previous reports have shown that rat, murine, and human skin and cultured fibroblasts possessed an effector system for 1,25(OH)2D3 (1)(2)(3)(4)(5) suggesting that these tissues could serve as targets for the hormonal form of vitamin D3. Moreover, it has been established that these cells were responsive to 1 ,25(OH)2D3…”

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Specific 1,25-Hydroxycholecalciferol Receptors and Stimulation of 25-Hydroxycholecalciferol- 24R Hydroxylase in Human Amniotic Cells

1987

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ABSTRACT. We have analyzed the la,25-dihydroxycholecalciferol [1,25(OH)2D3] receptor content of cultured cells from human amniotic fluid. Six cell lines were grown to confluence in a minimum essential medium containing 20% fetal calf serum. All had a normal karyotype, five were male and one was female. Hypertonic cytosol extracts were prepared by sonication followed by centrifugation at Previous reports have shown that rat, murine, and human skin and cultured fibroblasts possessed an effector system for 1,25(OH)2D3 (1-5) suggesting that these tissues could serve as targets for the hormonal form of vitamin D3. Moreover, it has been established that these cells were responsive to 1 ,25(OH)2D3by showing an unequivocal stimulation of 24-OHase (6, 7).

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Binding of 1,25‐dihydroxy‐[³H]vitamin D₃ in nuclear and cytosol fractions of whole mouse skin in vivo and in vitro

Cited by 25 publications

References 20 publications

Interaction of 1,25-Dihydroxyvitamin-D₃with Keratinocytes and Fibroblasts from Skin of Normal Subjects and a Subject with Vitamin-D-Dependent Rickets, Type II: A Model for Study of the Mode of Action of 1,25-Dihydroxyvitamin D₃*

Interaction of 1,25-Dihydroxyvitamin-D₃with Keratinocytes and Fibroblasts from Skin of Normal Subjects and a Subject with Vitamin-D-Dependent Rickets, Type II: A Model for Study of the Mode of Action of 1,25-Dihydroxyvitamin D₃*

Novel regulatory actions of 1α,25‐dihydroxyvitamin D₃ on the metabolism of polyphosphoinositides in murine epidermal keratinocytes

Specific 1,25-Hydroxycholecalciferol Receptors and Stimulation of 25-Hydroxycholecalciferol- 24R Hydroxylase in Human Amniotic Cells

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Binding of 1,25‐dihydroxy‐[3H]vitamin D3 in nuclear and cytosol fractions of whole mouse skin in vivo and in vitro

Cited by 25 publications

References 20 publications

Interaction of 1,25-Dihydroxyvitamin-D3with Keratinocytes and Fibroblasts from Skin of Normal Subjects and a Subject with Vitamin-D-Dependent Rickets, Type II: A Model for Study of the Mode of Action of 1,25-Dihydroxyvitamin D3*

Interaction of 1,25-Dihydroxyvitamin-D3with Keratinocytes and Fibroblasts from Skin of Normal Subjects and a Subject with Vitamin-D-Dependent Rickets, Type II: A Model for Study of the Mode of Action of 1,25-Dihydroxyvitamin D3*

Novel regulatory actions of 1α,25‐dihydroxyvitamin D3 on the metabolism of polyphosphoinositides in murine epidermal keratinocytes

Specific 1,25-Hydroxycholecalciferol Receptors and Stimulation of 25-Hydroxycholecalciferol- 24R Hydroxylase in Human Amniotic Cells

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Binding of 1,25‐dihydroxy‐[³H]vitamin D₃ in nuclear and cytosol fractions of whole mouse skin in vivo and in vitro

Interaction of 1,25-Dihydroxyvitamin-D₃with Keratinocytes and Fibroblasts from Skin of Normal Subjects and a Subject with Vitamin-D-Dependent Rickets, Type II: A Model for Study of the Mode of Action of 1,25-Dihydroxyvitamin D₃*

Interaction of 1,25-Dihydroxyvitamin-D₃with Keratinocytes and Fibroblasts from Skin of Normal Subjects and a Subject with Vitamin-D-Dependent Rickets, Type II: A Model for Study of the Mode of Action of 1,25-Dihydroxyvitamin D₃*

Novel regulatory actions of 1α,25‐dihydroxyvitamin D₃ on the metabolism of polyphosphoinositides in murine epidermal keratinocytes