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  • 1
    Electronic Resource
    Electronic Resource
    Differential effects of transforming growth factor-β1 and bone morphogenetic protein 4 on gene expression and differentiated function of preosteoblasts (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 155 (1993), S. 112-119 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Transforming growth factor-β (TGF-β) and bone morphogenetic protein 4 (BMP 4) are both able, under certain circumstances, to induce endochondral bone formation in vivo. This study compared the effects of TGF-β and BMP 4 on the gene expression of a retinoic acid (RA) responsive rat clonal preosteoblast cell line, UMR 201, as well as the way in which these proteins interact with RA in these cells. Both similarities as well as differences between the effects and mechanism of action of TGF-β1 and BMP 4 were demonstrated. TGF-β1 (0.1 ng/ml) strongly induced matrix gla protein (MGP) mRNA and increased the steady state osteonectin (ON) mRNA level. Cotreatment with TGF-β1 and RA did not result in a further increase in MGP mRNA expression. In contrast, BMP 4 alone had no influence on MGP or ON mRNA expression but it significantly enhanced the RA induction of MGP mRNA. Pro-α(1) (l) collagen mRNA was increased by TGF-β1 (1 ng/ml) and BMP 4 (50 ng/ml). The addition of either TGF-β1 or BMP 4 together with RA resulted in a further increase in pro-α1(l) collagen mRNA levels. Both RA and TGF-β1, but not BMP 4, increased the transcriptional rate of the pro-α 1(l) collagen gene. TGF-β1 reduced the constitutive as well as RA-induced expression of osteopontin (OP) mRNA while BMP 4 reduced only the constitutive expression of OP mRNA. RA increased the transcriptional rate of the OP gene. Since the responses of UMR 201 cells to these structurally related factors were not identical, the results lend support to the concept that the coordinated expression of members of the TGF-β1 superfamily may be necessary to control the progression of specific cell types through their differentiation pathways. © 1993 Wiley-Liss, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041550115
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  • 2
    Electronic Resource
    Electronic Resource
    Regulation of Na+/glucose cotransporter (SGLT1) mRNA in LLC-PK1 cells (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 158 (1994), S. 506-512 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The porcine kidney epithelial cell line LLC-PK1 expresses a sodium-coupled glucose cotransporter (SGLT1) together with other differentiation markers of renal proximal tubule such as trehalase and γ-glutamyltranspeptidase. Expression is regulated by cell density and exogenous differentiation inducers such as hexamethylene bisacetamide (HMBA). Northern blot and PCR analysis of clonal cell populations indicated SGLT1 mRNA was not detectable in subconfluent cultures, but 2.2 and 3.9 kb SGLT1 mRNA species appeared after cell confluence, accompanying expression of the transport activity. SGLT1 mRNA levels were significantly increased after treatment of confluent cultures with HMBA, paralleling increases in the transport activity and immunodetectable 75 kD cotransporter subunit. SGLT1 mRNA was also increased after treatment of cultures with the cyclic AMP phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX), an inducer of Na+/glucose cotransport activity. The 3.9 kb SGLT1 transcript showed the largest increase after either HMBA or IBMX treatment. HMBA treatment also resulted in increased mRNA levels of two other differentiation markers-trehalase and γ-glutamyltranspeptidase. By contrast, trehalase and γ-glutamyltranspeptidase mRNA levels were not increased by IBMX. Regulation of Na+/glucose symporter expression by either cell density, cyclic AMP elevation, or differentiation inducer treatment occurs, at least in part, at the level of SGLT1 mRNA and can be dissociated from regulation of other differentiation markers. © 1994 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041580315
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  • 3
    Electronic Resource
    Electronic Resource
    Regulation of Cu,Zn-superoxide dismutase in bovine pulmonary artery endothelial cells (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 153 (1992), S. 491-497 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: To evaluate the regulation of endothelial cell Cu,Zn-SOD, we have exposed bovine pulmonary artery endothelial cells in culture to hyperoxia and hypoxia, second messengers or related agonists, hormones, free radical generating systems, endotoxin, and cytokines and have measured Cu,Zn-SOD protein of these cells by an ELISA developed in our laboratory. Control preconfluent and confluent cells in room air contained 196 ± 18 ng Cu,Zn-SOD/106 cells. A23187 (0.33 μM), for skolin (10 μM), isobutylmethylxanthine (0.1 mM), dexamethasone (1 μM), triiodothyronine (1 μM) and retinoic acid (1 μM) failed to alter this level of Cu,Zn-SOD. Exposure to anoxia and hyperoxia both elevated the level -1.5-2.0-fold over 20% oxygen-exposed controls at 48-72 hr. Similarly, exposures to glucose oxidase (0.0075 units/ml), menadione (12.5 μM), xanthine-xanthine oxidase (10 μM, 0.03 units/ml) and H2O2 (0.0005%) increased the level up to two-threefold over controls at 24-48 hr. Lipopolysaccharide, TGFβ1, TNFα, and II-1 also increased levels of cellular Cu,Zn-SOD, but only in proliferating cells. II-2, II-4, interferon-γ, and GM-CSF had no effect on Cu,Zn-SOD. All treatments that elevated SOD resulted in inhibition of cellular growth, but decreased growth of cells at confluence alone was not associated with increased Cu,Zn-SOD. We propose from these studies that Cu,Zn-SOD of endothelial cells is not under conventional second messenger or hormonal regulation, but that up-regulation of the enzyme is associated with (and perhaps stimulated by) free-radical or oxidant production that also may be influenced by availability of certain cytokines under replicating conditions. © 1992 Wiley-Liss, Inc.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041530308
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  • 4
    Electronic Resource
    Electronic Resource
    A quantitative analysis of mitochondria during fetal mouse oogenesis (1988)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 195 (1988), S. 225-234 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A quantitative analysis of mitochondrial populations during the meiotic prophase of mouse oogenesis was carried out. The mean absolute area occupied by mitochondria and the mean number of mitochondria per cell increases in a linear fashion from pachytene through dictyate. The mean area occupied by mitochondria increases at pachytene and thereafter. Both small and large aggregations of mitochondria are seen, particularly during the later stages of prophase. Vacuolated mitochondria are present from preleptotene through pachytene. Mitochondria show major dynamic changes throughout fetal mouse oogenesis, which may suggest significant functional activities yet to be elucidated.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1051950208
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  • 5
    Electronic Resource
    Electronic Resource
    Multitude of inverted repeats characterizes a class of anchorage sites of chromatin loops to the nuclear matrix (1993)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 53 (1993), S. 1-12 
    Details
    ISSN: 0730-2312
    Keywords: cloning ; nuclear matrix ; MARs ; origins of replication ; palindromes (inverted repeats) ; transcriptional enhancers ; chromatin domains ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: In order to understand the nature of DNA sequences that organize chromatin into domains or loops, we have cloned the nuclear matrix DNA (1.7% of the total DNA) from human myelogeneous leukemia cells in culture. Nuclear matrix is formed by interactions between specific stretches of DNA of about 0.1 to 5.0 kb with protein transcription factors, nuclear enzymes, and structural proteins. Nuclear matrix is believed to be the exclusive nuclear microenvironment in which initiation of DNA replication, transcription, and repair take place. The matrix attachment regions (MARs) of DNA have transcriptional enhancer activity, harbor the origins of replication of the human genome, and define the borders between neighbouring chromatin loops. In this study we report the sequence of the human MAR fragment 19.2 of a size of 542 bp. Hum. MAR 19.2 is composed of TG-, CA-, CT-, and GA-rich blocks and shows 8 perfect and imperfect inverted repeats. Thus, we have identified a novel class of MARs with sequence characteristics divergent from the AT-rich class of MARs. The inverted repeats of the 19.2 sequence might be stabilized into their cruciform configuration by torsional strain and by specific transcription/replication protein factors. This MAR might function in the initiation of replication of the flanking chromatim domain and in the regulation of the transcriptional activity of the gene(s) that reside in this domain.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240530102
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  • 6
    Electronic Resource
    Electronic Resource
    Identification of a core motif that is recognized by three members of the HMG class of transcriptional regulators: IRE-ABP, SRY, and TCF-1α (1992)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 48 (1992), S. 129-135 
    Details
    ISSN: 0730-2312
    Keywords: insulin ; gene transcription ; GAPDH ; hormone regulation ; differentiation-dependent gene expression ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Insulin induces glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene transcription in part by regulating one or more proteins that bind a cis-acting element, IRE-A. We have recently cloned a protein, IRE-ABP, that binds the IRE-A element. IRE-ABP is a member of the HMG class of transcriptional regulators and is 67% identical within its HMG box domain to the candidate gene for the testis-determining factor, SRY. IRE-ABP and SRY share binding specificity for the IRE-A motif. This sequence is also highly conserved with a core motif, 5′-Py-ctttg(a/t)-3′, contained in T-cell specific genes that have high affinity for TCF-1α, another member of the HMG class of transcriptional regulators. Thus, diverse members of the HMG family interact with similar nucleotide sequences to regulate expression of genes that initiate and maintain the differentiated phenotype. We have found this core motif in the upstream region of many genes that are positively and negatively regulated by insulin. These observations suggest that IRE-ABP or a related family member may coordinate the expression of these genes. The HMG family of proteins has diverse functions ranging from the regulation of differentiation and mating type in yeast to the regulation of tissue-and species-specific gene expression in mammals. Insulin regulates GAPDH gene transcription in a tissue-specfic manner. We propose that members of the IRE-ABP family play an important role in controlling tissue specificity of the insulin response.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240480204
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  • 7
    Electronic Resource
    Electronic Resource
    rHox: A homeobox gene expressed in osteoblastic cells (1995)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 59 (1995), S. 486-497 
    Details
    ISSN: 0730-2312
    Keywords: homeobox gene ; rHox ; rat ; osteocalcin ; collagen I α 1 ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Homeodomain proteins are characterized by a conserved domain with a helix-turn-helix motif. These proteins act as regulatory factors in tissue differentiation and proliferation. However, their role in the regulation of osteoblast differentiation is unknown. In this study we have identified and characterized a homeobox gene in osteoblast-like cells. This gene, termed rHox, was isolated from a cDNA library derived from rat osteoblast-like cells. The nucleotide sequence of the 1,375 base pair (bp) cDNA contains a noncoding leader sequence of 329 bp, a 735 bp open reading frame, and 312 bp of 3′ noncoding sequence. Sequence comparison demonstrates that rHox is identical to the mouse Pmx gene (also called MHox) at the amino acid level and 90% homologous at the nucleotide level. Both Southwestern blotting and gel shift analyses indicate that rHox has potential to bind both the collagen l α 1 and the osteocalcin promoters. Transfection experiments using an rHox expression vector showed a strong repression of target promoter activity, regardless of whether the target promoters contained homeodomain binding reponse elements. These data suggest that rHox is a potent negative regulator of gene expression, although the specific role of rHox in bone gene regulation remains to be determined. © 1995 Wiley-Liss, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240590409
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  • 8
    Electronic Resource
    Electronic Resource
    Transforming activity of human nasopharyngeal carcinoma DNA (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 129 (1986), S. 21-26 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: NIH 3T3 cells were transfected with the DNAs from biopsy specimens of human nasopharyngeal carcinoma (NPC, EBV DNA positive) using calcium phosphate precipitation method. The malignant, transformed foci of NIH 3T3 cells have been observed and cloned. The hybridization of transfectant DNA digested by EcoRI with total human leukocyte DNA as probe was performed. The strong signal of smear comparing with NIH 3T3 DNA as control was observed. It was implied that the putative human transforming sequences had been integrated into transformed cells. Employing soft agar culture, the transformed cells can grow and form cell colonies. Following transfer, the foci were able to grow and adhere to a glass wall. These cells were easily agglutinated by con A. The cloned foci have been inoculated into nude mice with the formation of highly malignant sarcomas. In preliminary experiments for characterizing the transforming sequences, Ha-ras and Blym 1 were found in transfectants derived from one of the NPC DNA samples. It is implicated that these two oncogenes might be responsible for the acquisition of malignant phenotypic character of some human NPC. The further identification of oncogenes in NPC is currently in progress.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041290406
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  • 9
    Electronic Resource
    Electronic Resource
    Abnormal proteins as the trigger for the induction of stress responses: Heat, diamide, and sodium arsenite (1988)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 136 (1988), S. 411-420 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Thermotolerance and synthesis of heat shock proteins are induced in cells in response to a variety of environmental stresses. We examined the suggestion of Hightower (1980) that modifications of intracellular proteins may be the triggering event that induces heat shock protein synthesis and thermotolerance. We did so by modifying cellular proteins, using diamide, a sulfhydryl oxicizing agent, and dithio-bis (succinimidyl propionate), an agent that cross-links bifunctional amino groups. Both of these agents induced heat shock proteins and thermotolerance in CHO (HA-1) cells. Furthermore, we observed cross-resistance and self-tolerance with three seemingly unrelated stimuli (diamide, heat, and sodium arsenite). This observation suggests that the induction of protective responses to these stimuli is mediated by a common mechanism. The results support the hypothesis that production of abnormal proteins by various stresses induces the stress responses as well as tolerance.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041360304
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  • 10
    Electronic Resource
    Electronic Resource
    The kinetics of the first wave of spermatogenesis in the newborn male mouse (1986)
    New York, NY : Wiley-Blackwell
    Gamete Research 14 (1986), S. 245-254 
    Details
    ISSN: 0148-7280
    Keywords: mammalian ; germ cell ; development ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A combination of autoradiography and air-dried techniques was used to calculate the duration of the major meiotic stages in the first wave of spermatogenesis in the newborn mouse. The data indicated that the entry into meiosis occurred asynchronously over 2 days, and the time required for each stage and the total cycle was constant. These time intervals were nearly identical with those estimated for adult animals in the present study and by other authors.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120140308
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