Evidence for receptor-mediated mineralocorticoid action in rat osteoblastic cells

Agarwal, M.K.; Mirshahi, Farrokh; Mirshahi, Massoud; Bracq, S.; Chentoufi, Jamila; Hott, M.; Jullienne, A.; Marie, Pierre J.

doi:10.1152/ajpcell.1996.270.4.c1088

Cited by 24 publications

(31 citation statements)

References 28 publications

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“…Thus, lymphocytes are endowed with the MCR although its functional relevance there remains unknown. 4 A comparable study with murine osteoblastic cells revealed altered growth response with steroids targeted for specificity to the MCR 18 and a similar situation could well be envisioned with hematopoietic cells.…”

Section: Introductionmentioning

confidence: 79%

“…To assess multiplication, 80 000 cells were transferred to 12-well tissue culture plates (six per sample), exposed to the desired combination of steroids dissolved in ethanol (1% final) in a serum-free medium for 3 days, and finally counted manually with the aid of a hematocytometer. 18 The condensation of chromatin was measured by a fluorometric procedure with the aid of Hoechst 33342 (Germany). For this purpose, 4 × 10 4 cells were grown to 80-90% confluence in 96-well tissue culture plates and reincubated in RPMI 1640 supplemented with glutamine, antibiotics, and 2% FCS.…”

Section: Cell Culturementioning

confidence: 99%

“…Both preparations were kept frozen at −80°C until use. The protein content was estimated by the method of Lowry et al, 18 as in previous studies.…”

Section: Cell Extractmentioning

confidence: 99%

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Demonstration of the mineralocorticoid hormone receptor and action in human leukemic cell lines

Mirshahi

Golestaneh

et al. 2000

Leukemia

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We studied the expression of the mineralocorticoid receptor (MCR), and of the amiloride-sensitive sodium channel (ASSC) regulated by the MCR, in human leukemic cell lines. Cell extracts from TF1 (proerythroblastic), HEL (human erythroblastic leukemia) and U937 (myeloblastic) cell line were positive for the ASSC, as a 82 kDa band in Western blots developed with the aid of a polyclonal antibody raised against the peptide QGLGKGDKREEQGL, corresponding to the region 44-58 of the ␣ subunit of the epithelial sodium channel (ENaC) cloned from rat colon, linked to KLH. The polyclonal antibody against the MCR revealed a single band of about 102 kDa in extracts from HEL and TF1 cells. The immunofluorescent labelling of the MCR in all cell lines showed a nucleocytoplasmic localization of the receptor but the ASSC was exclusively membrane-bound and these results were confirmed by confocal microscopy. The expression of the MCR in the HEL cells was evident as a predicted band of 843 bp (234 amino acids) in electrophoresis of the PCR product obtained after total RNA had been reverse transcribed and then amplified using the primers 5′-AGGCTAC-CACAGTCTCCCTG-3′ and 5′-GCAGTGTAAAATCTCCAGTC-3′ (sense and antisense, respectively). The ENaC was similarly evident with the aid of the primers 5′-CTGCCTTTATG GATGATGGT-3′ (sense) and 5′-GTTCAGCTCGAAGAAGA-3′ (antisense) as a predicted band of 520 bp. In both cases, 100% identity was observed between the sequences of the PCR products compared to those from known human sources. The multiplication of the HEL cells was influenced by antagonists (RU 26752, ZK 91587) targeted for specificity to the MCR and this was selectively reversed by the natural hormone aldosterone. These steroids also provoked chromatin condensation in the HEL population. These permit new and novel possibilities to understand the pathobiology of human leukemia and to delineate sodium-water homeostasis in nonepithelial cells. Leukemia (2000) 14, 1097-1104.

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Section: Introductionmentioning

confidence: 79%

Section: Cell Culturementioning

confidence: 99%

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Demonstration of the mineralocorticoid hormone receptor and action in human leukemic cell lines

Mirshahi

Golestaneh

et al. 2000

Leukemia

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“…The cells were trypsinized and counted with the aid of either a cell counter or a hematocytometer [16,17,19].…”

Section: Cell Multiplicationmentioning

confidence: 99%

“…The anti-MCR serum was raised by immunizing rabbits with the protein purified biochemically from rat kidney in the presence of RU 26752 that binds specifically to this receptor [11,19]; the IgG fraction was obtained by fractionating the serum through a diethylaminoethanol-Trisacryl column [16,17].…”

Section: Chemicals and Reagentsmentioning

confidence: 99%

Mineralocorticoid Hormone Signaling Regulates the ‘Epithelial Sodium Channel’ in Fibroblasts from Human Cornea

Mirshahi

Golestaneh

et al. 2000

Ophthalmic Res

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We investigated the regulation of sodium absorption by steroid hormones in embryologically diverse cells from the human eye. A cell extract from human corneal fibroblasts was positive for both the epithelial sodium channel (ENaC) and the mineralocorticoid receptor (MCR) as 82- to 85-kD and 102-kD bands, respectively, by the Western blot technique. In fluorescent, confocal and electron microscopy, the MCR was revealed as a nucleocytoplasmic protein, whereas the ENaC was almost exclusively membrane bound; both appeared aligned along actin filaments of corneal keratocytes, and both were widely colocalized in various cell types of human cornea in situ. Following reverse transcription and amplification of total RNA isolated from corneal fibroblasts, the ENaC and MCR genes in the PCR product were evident as predicted bands of 520 and 843 bp, respectively, whose sequence exhibited 100% identity with those from known human sources. The multiplication of corneal fibroblasts was influenced by both the MCR-specific antagonist RU 26752 and the natural hormone aldosterone, and these steroids also stimulated protein phosphorylation. In quantitative PCR, both the basal and aldosterone-induced levels of ENaC were diminished by the MCR-specific antagonist ZK 91587. Consequently, the ocular sodium channel appears to be regulated by steroid signalling in cells of diverse embryological origins, contrary to the existing notions where (a) this process would be limited exclusively to the epithelial cells and (b) ocular sodium transport would be regulated via the Na⁺-K⁺-ATPase in the basolateral membrane.

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Effects of glucocorticoids and mineralocorticoids on proliferation and maturation of human peripheral blood stem cells

Grafte-Faure¹,

Leveque²,

Vasse³

et al. 1999

Am. J. Hematol.

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It has been shown that hematopoietic progenitors can be expanded ex vivo in the presence of various cytokine combinations. Glucocorticoids (GC) are involved in the self-renewal of erythroid progenitors in chicken. To see whether GC have a similar effect on hematopoiesis in humans, CD34(+) peripheral blood stem cells were cultured in serum free medium in the presence of a GC, triamcinolone acetonide. However, our results demonstrate an inhibition of both erythroid and granulocyte-macrophage (GM) proliferation and a modification of erythroid colony morphology. Furthermore, RU38486 (Mifepristone), a potent GC antagonist, was unable to reverse the inhibitory effect of triamcinolone acetonide. We also identified and characterized another steroid subfamily, the mineralocorticoid (MC) subfamily, in human PB CD34(+) cells. The MC, aldosterone, significantly enhanced GM colony formation and diminished the erythroid colony number. Neither of effects were inhibited by ZK91587, an antagonist specific to the MC receptor (MCR). In contrast, ZK91587 reversed the stimulatory effect of deoxycorticosterone on GM colony formation. Cytoplasmic staining for MCR was observed in CD34(+) cells incubated with a polyclonal antiserum raised against human MCR. To our knowledge, this is the first demonstration of the presence of MCR in human PB CD34(+) cells.

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Evidence for receptor-mediated mineralocorticoid action in rat osteoblastic cells

Cited by 24 publications

References 28 publications

Demonstration of the mineralocorticoid hormone receptor and action in human leukemic cell lines

Demonstration of the mineralocorticoid hormone receptor and action in human leukemic cell lines

Mineralocorticoid Hormone Signaling Regulates the ‘Epithelial Sodium Channel’ in Fibroblasts from Human Cornea

Effects of glucocorticoids and mineralocorticoids on proliferation and maturation of human peripheral blood stem cells

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