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ESE-3, a novel member of an epithelium-specific Ets transcription factor subfamily, demonstrates different target gene specificity from ESE-1

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第4期275卷 2986-2998页

作者： Kas, K Finger, E Grall, F Gu, XS Akbarali, Y Boltax, J Weiss, A Oettgen, P Kapeller, R Libermann, TA Harvard Univ Inst Med Beth Israel Deaconess Med Ctr New England Baptist Bone & Joint InstDept Med Boston MA 02115 USA Harvard Univ Sch Med Boston MA 02115 USA Katholieke Univ Leuven Ctr Human Genet Mol Oncol Lab B-3000 Louvain Belgium Flanders Interuniv Inst Biotechnol B-3000 Louvain Belgium Millennium Pharmaceut Inc Cambridge MA 02139 USA

Most cancers originate as a result of aberrant gene expression in mainly glandular epithelial tissues leading to defects in epithelial cell differentiation. The latter is governed by distinct sets of transcriptional regulators. Here we report the characterization of epithelium-specific Ets factor, family member 3 (ESE-3), a novel member of the ESE subfamily of Ets transcription factors. ESE-3 shows highest homology to two other epithelium restricted Ets factors, ESE-1 and ESE-2. ESE-3, like ESE-1 and ESE-2, is exclusively expressed in a subset of epithelial cells with highest expression in glandular epithelium such as prostate, pancreas, salivary gland, and trachea. A potential role in branching morphogenesis is suggested, since ESE-3 transactivates the c-MET promoter via three high affinity binding sites. Additionally, ESE-3 binding to DNA sequences in the promoters of several glandular epithelium-specific genes suggests a role for ESE-3 in later stages of glandular epithelium differentiation. Although ESE-3 and ESE-1 bind with similar affinity to various Ets binding sites, ESE-3 and ESE-1 differ significantly in their ability to transactivate the promoters containing these sites. Our results support the notion that ESE-1, ESE-2, and ESE-3 represent a unique epithelium-specific subfamily of Ets factors that have critical but distinct functions in epithelial cell differentiation and proliferation.

关键词：衔接蛋白质类膜泡运输氨基酸序列碱基序列载体蛋白质类/代谢细胞培养的克隆分子 DNA 互补上皮/代谢基因打靶分子序列数据多基因族种系发生启动区遗传原癌基因蛋白质类/遗传学原癌基因蛋白质类/代谢原癌基因蛋白质c-ets 序列同源性氨基酸转录因子/遗传学转录因子/代谢转录激活动物人类

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Subgroup of reproductive functions of progesterone mediated by progesterone receptor-B isoform

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SCIENCE 2000年第5485期289卷 1751-1754页

作者： Mulac-Jericevic, B Mullinax, RA DeMayo, FJ Lydon, LP Conneely, OM Baylor Coll Med Dept Mol & Cellular Biol Houston TX 77030 USA

Progesterone regulates reproductive function through two intracellular receptors, progesterone receptor-A (PR-A) and progesterone receptor-B (PR-B), that arise from a single gene and function as transcriptional regulators of progesterone-responsive genes. Although in vitro studies show that PR isoforms can display different transcriptional regulatory activities, their physiological significance is unknown. By selective ablation of PR-A in mice, we show that the PR-B isoform modulates a subset of reproductive functions of progesterone by regulation of a subset of progesterone-responsive target genes. Thus, PR-A and PR-B are functionally distinct mediators of progesterone action in vivo and should provide suitable targets for generation of tissue-selective progestins.

关键词：细胞分裂/药物作用交叉遗传胚胎植入上皮细胞/细胞学上皮细胞/药物作用上皮/药物作用上皮/代谢雌二醇/药理学基因表达调控乳腺动物/细胞学乳腺动物/药物作用小鼠基因敲除卵巢切除术排卵孕酮/药理学孕酮/生理学蛋白质亚型受体孕酮/遗传学受体孕酮/生理学生殖子宫/细胞学子宫/药物作用子宫/代谢子宫/生理学动物女(雌)性男(雄)性小鼠

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DIFFERENTIAL EFFECTS OF VASOPRESSIN ON WATER, CALCIUM AND LYSOSOMAL ENZYME CONTENTS OF MITOCHONDRIA-RICH AND LYSOSOME-RICH (GRANULAR) EPITHELIAL-CELLS ISOLATED FROM BULLFROG URINARY-BLADDER

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MOLECULAR AND CELLULAR ENDOCRINOLOGY 1976年第2期4卷 89-106页

作者： PIETRAS, RJ NAUJOKAITIS, PJ SZEGO, CM UNIV CALIF DEPT BIOL LOS ANGELES CA 90024 USA UNIV CALIF MOLEC BIOL INST LOS ANGELES CA 90024 USA

Treatment of bullfrog urinary bladder with arginine vasopressin (AVP) elicited a dose-dependent increase in the basal movement of H2O and Na across isolated tissues. Epithelial cells from the mucosal surface and incubated with 10 mU [milliunits] AVP/ml for 30 min retained a greater amount of intracellular H2O and Ca than cells not treated th hormone. Epithelial cells were further separated into 2 major fractions by density gradient centrifugation;cells damaged during these manipulations were separated from viable cells and discarded. Morphological examination of the 2 respective fractions indicated that they largely consisted of mitochondria-rich (MR) and granular (G) cell types which line the lumen of bullfrog bladder. The Ca content of MR cells averaged 25% greater than that of G-type cells. G cells had a higher content of the characteristic lysosomal hydrolases, acid phosphatase and cathepsin B1 than that found in MR cells. Incubation of G cells with AVP elicited significant increments in H2O and Ca contents and extracellular release of lysosomal enzymes as compared to untreated cells. Among MR cells treated with AVP, Cell Ca declines slightly, but no significant increase in H2O content of extracellular hydrolase activity was detected in comparison with paired control cells. The physiological significance of acid proteinase release from G cells treated with AVP was evaluated in experiments with intact bladder. Proteinase inhibitors which suppress the activity of cathepsin B1 selectively antagonized the action of hormone on H2O permeation. Alterations in the Ca and lysosomal hydrolase activity associated with G cells exposed to AVP may contribute to the hormone-induced H2O flow observed across the intact epithelium.

关键词：酸性磷酸酶/代谢无尾目精氨酸加压素/药理学身体水份/代谢钙/代谢组织蛋白酶类/代谢细胞质颗粒/代谢细胞质颗粒/超微结构上皮细胞上皮/药物作用上皮/代谢上皮/超微结构溶酶体/药物作用溶酶体/代谢显微镜检查电子线粒体/药物作用线粒体/代谢线粒体/超微结构牛蛙膀胱/药物作用膀胱/代谢血管升压素类/类似物和衍生物动物

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ROLE OF EPITHELIAL ARCHITECTURE AND INTRACELLULAR METABOLISM IN PROLINE UPTAKE AND TRANSTUBULAR RECLAMATION IN PRO-RE MOUSE KIDNEY

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 1975年第4期72卷 1431-1435页

作者： SCRIVER, CR MCINNES, RR MOHYUDDIN, F MCGILL UNIV DEPT PEDIATMONTREALQUEBECCANADA MCGILL UNIV MONTREAL CHILDRENS HOSPRES INSTDEBELLE LAB BIOCHEM GENETMONTREALQUEBECCANADA MCGILL UNIV DEPT BIOLMONTREALQUEBECCANADA

The homozygous PRO/Re mouse has less than 1 percent of the very high proline oxidase activity that characterizes normal kidney cortex. In PRO/Re mouse the endogenous proline concentration is eight times normal in plasma and four times normal in kidney cortex cell, but 50 times normal in urine. The integrity of the membrane transport systems for proline uptake at the antiluminal surface of absorbing epithelium is retained in PRO/Re kidney, as determined by the slice method. Clearance studies in vivo under steady-state conditions indicate that the integrity of the luminal uptake system shared by glycine and proline, and serving proline absorption, is also intact. The exaggerated renal clearance of proline in PRO/Re mice (50 times normal) is explained when its raised intracellular concentration, caused by impaired proline oxidation, is considered. Backflux into urine flowing down the nephron will occur under these conditions, thus impairing net reclamation of proline in PRO/Re kidney. The findings reveal that membrane transport and intracellular metabolism of a substrate are, indeed, independent functions, but that metabolism of a substance can influence its transcellular transport.

关键词：生物转运上皮细胞上皮/代谢上皮/超微结构肾小球滤过率肾皮质/代谢肾小管/代谢动力学小鼠近交系脯氨酸/代谢种特异性动物人类小鼠大鼠

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INHIBITION OF PALATAL EPITHELIAL-CELL DEATH BY ALTERED PROTEIN-SYNTHESIS

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DEVELOPMENTAL BIOLOGY 1976年第1期54卷 135-145页

作者： PRATT, RM GREENE, RM NIDR DEV BIOL & ANOMALIES LAB BETHESDA MD 20014 USA

Programmed cell death occurs in a defined region of the secondary palatal epithelium (medial edge) during its development in vivo or in vitro. The effects of various inhibitors of macromolecular synthesis, particularly glycoprotein synthesis, on the death of palatal medial-edge epithelial cells in vitro was studied. Rat palatal shelves explanted on gestational day 15 and cultured singly showed medial-edge cell degeneration after 48 h, whereas in the presence of 6-diazo-5-oxo-L-norleucine (DON), 2-deoxyglucose (DOG) or cycloheximide, cell death was prevented. This was in contrast to the lack of inhibition of cell death in the presence of actinomycin D or cytosine arabinoside. Selectively adding metabolites which bypass the effect of DON on various synthetic pathways demonstrated that the inhibition of cell death by DON was most likely due to a block in glucosamine formation. The synthesis and activity of various lysosomal enzymes were not affected by DON, whereas a marked decrease was observed with both cycloheximide and DOG. Palatal epithelial cell death may not be a passive event, but an active process requiring the programmed synthesis of specific proteins.

关键词：细胞存活放线菌酮/药理学阿糖胞苷/药理学更生霉素/药理学重氮亮氨酸/药理学上皮细胞上皮/代谢糖蛋白类/生物合成器官培养技术腭/胚胎学时间因素动物女(雌)性大鼠

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Activation of cystic fibrosis transmembrane conductance regulator in rat epididymal epithelium by genistein

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BIOLOGY OF REPRODUCTION 2000年第1期62卷 143-149页

作者： Leung, GPH Wong, PYD Chinese Univ Hong Kong Fac Med Dept Physiol Shatin New Territories Peoples R China

The effect of genistein on anion secretion via cystic fibrosis transmembrane conductance regulator (CFTR) in cultured rat cauda epididymal epithelia was studied by short-circuit current (Isc) technique. Genistein added apically stimulated a concentration-dependent rise in Isc due to Cl- and HCO3- secretion. The genistein-induced Isc was observed in basolaterally permeabilized monolayers, suggesting that the Isc response was mediated by the apical anion channel. The response could be blocked by the nonspecific Cl- channel blocker, diphenylamine-2-carboxylate (DPC), but not by the Ca2+-activated Cl- channel blocker, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). Genistein did not increase intracellular cAMP, but H-89, a protein kinase A inhibitor, completely abolished the Isc response to genistein. Moreover, pretreatment of the tissues with MDL-12330A, an adenylate cyclase inhibitor, markedly attenuated the Isc response to genistein, but the response was restored upon the addition of exogenous cAMP. Ca2+, protein kinase C, tyrosine kinase, and protein phosphatase signalling pathways were not involved in the action of genistein. It is speculated that genistein stimulates anion secretion by direct interaction with CFTR. This requires a low level of phosphorylation of CFTR by basal protein kinase A activity. If is suggested that genistein may provide therapeutic benefit to male infertility associated with cystic fibrosis.

关键词： 4 4'-二异硫氰基苯-2 2'-二磺酸/药理学腺苷酸环化酶/拮抗剂和抑制剂邻氨基苯甲酸类/药理学碳酸氢盐类/代谢细胞培养的氯化物通道/拮抗剂和抑制剂氯化物/代谢环AMP/代谢囊性纤维化跨膜传导调节因子/代谢导电性酶抑制剂/药理学附睾/代谢上皮/代谢染料木黄酮/药理学蛋白激酶抑制剂大鼠 Sprague-Dawley 毒胡萝卜内酯/药理学动物男(雄)性大鼠

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Immunohistochemical localization of thrombomodulin in the stratified epithelium of the rat is restricted to the keratinizing epidermis

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HISTOCHEMISTRY AND CELL BIOLOGY 1999年第6期112卷 437-442页

作者： Daimon, T Nakano, M Teikyo Univ Sch Med Dept Anat Itabashi Ku Tokyo 1738605 Japan Mitsubishi Gas Chem Co Niigata Res Lab Niigata 95031 Japan

The expression and function of thrombomodulin (TM), an endothelial cofactor protein for thrombin-mediated protein C activation, in the epithelium are not fully characterized. This report describes the distribution and localization of TM in the various types of epithelia in the rat by light and electron microscopic immunocytochemistry. TM showed a limited distribution and was expressed by the keratinizing stratified epithelia of the skin, tongue, and esophagus, but was not present on the non-keratinizing epithelia of the vagina, ureter, trachea, stomach, or gut. An identical pattern of TM expression was seen in mucocutaneous junctions, transitional zones from a non-keratinizing stratified epithelium to a keratinizing epithelium at the edge of the eyelid and in the anal canal. As the keratinization of the stratified epithelia proceeded, the staining intensity increased in the transitional zones. Within the keratinizing stratified epithelia, TM staining was limited to the keratinocytes of the spinous layer, the spinous cells. The subcellular localization of TM on the spinous cells was restricted to the plasma membrane facing the intercellular spaces. TM was not detectable on the desmosomes or the two membranes making up the junction, presumably the nexus. The functional significance of TM in keratinizing epithelia is discussed.

关键词：细胞膜/代谢表皮/代谢表皮/超微结构上皮/代谢上皮/超微结构免疫组织化学角蛋白细胞/代谢角蛋白细胞/超微结构显微镜检查免疫电子大鼠 Wistar 血栓调节蛋白/代谢动物女(雌)性男(雄)性大鼠

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Immunohistochemical localization of protein 3-nitrotyrosine and S-nitrosocysteine in a murine model of inhaled nitric oxide therapy

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PEDIATRIC RESEARCH 2000年第6期47卷 798-805页

作者： Lorch, SA Foust, R Gow, A Arkovitz, M Salzman, AL Szabo, C Vayert, B Geffard, M Ischiropoulos, H Childrens Hosp Philadelphia Stokes Res Inst Abramson Ctr 416D Dept Neonatol Philadelphia PA 19104 USA Childrens Hosp Med Ctr Div Surg Cincinnati OH 45229 USA Univ Bordeaux ENSCPB Lab PIOM EPHE Bordeaux France

Inhaled nitric oxide (INO) therapy is currently used clinically to selectively dilate the pulmonary vasculature and to help treat persistent pulmonary hypertension and bronchopulmonary dysplasia in the neonate. However, in the presence of oxygen or superoxide, nitric oxide forms potentially harmful reactive nitrogen species. Using an experimental mice model, we examined the effects of concurrent hyperoxia and INO on protein tyrosine nitration and cysteine S-nitrosylation in pulmonary tissue. Data showed enhanced S-nitrotyrosine staining within the airway epithelium and alveolar interstitium of mice lungs treated with hyperoxia, which did not increase significantly with INO administration. Within the alveolar interstitium, 3-nitrotyrosine staining was localized to macrophages. S-Nitrosocysteine staining in airway epithelium was significantly enhanced with INO administration regardless of oxygen content. These data suggest that the formation of protein S-nitrosocysteine is the major protein modification during administration of INO.

关键词：投药吸入半胱氨酸/类似物和衍生物半胱氨酸/代谢上皮/代谢免疫组织化学肺/代谢一氧化氮/投药和剂量一氧化氮/治疗应用亚硝基化合物/代谢 S-亚硝基硫醇酪氨酸/类似物和衍生物酪氨酸/代谢动物女(雌)性小鼠

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Increased expression of the type 1 growth factor receptor family in the conjunctival epithelium of patients with keratoconjunctivitis sicca

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AMERICAN JOURNAL OF OPHTHALMOLOGY 2000年第4期129卷 472-480页

作者： Liu, ZG Carvajal, M Carraway, CAC Carraway, KL Pflugfelder, SC Bascom Palmer Eye Inst Ocular Surface & Tear Ctr Miami FL 33136 USA Univ Miami Sch Med Dept Cell Biol & Anat Miami FL 33101 USA Univ Miami Sch Med Dept Biochem & Mol Biol Miami FL 33101 USA Sun Yatsen Univ Med Sci Zhongshan Ophthalm Ctr Guangzhou Peoples R China

PURPOSE: To investigate the expression of type 1 growth factor receptors (epidermal growth factor receptor, ErbB2, and ErbB3) in the conjunctival epithelium of patients with keratoconjunctivitis sicca. METHODS: Immunofluorescent staining and Western blotting were performed to grade the level of expression of the epidermal growth factor receptor ErbB2, and ErbB3 in conjunctival epithelial impression cytologies taken from both eyes of seven normal subjects and 22 patients with keratoconjunctivitis sicca. RESULTS: Epidermal growth factor receptor staining was observed in a greater percentage of keratoconjunctivitis sicca than normal samples (P < .05), ErbB2 and ErB3 staining in the apical conjunctival epithelium was observed in both groups, but stronger ErbB2 and ErbB3 staining was noted in keratoconjunctivitis sicca conjunctival samples (P < .05). The relative levels of expression of these receptor proteins on immunoblots were consistent with immunofluorescent staining. On immunoblots, epidermal growth factor receptor protein was detected in 50% of keratoconjunctivitis sicca samples, but none of the normal samples (P < .025). The expression of ErbB2 and ErbB3 on immunoblots was also greater in the keratoconjunctivitis sicca samples (P < .05). Immunofluorescent staining scores for these receptors were correlated with conjunctival lissamine green staining scores (r = .574, P < .01 for epidermal growth factor receptor;r = .620, P < .0025 for ErbB2;r = .502, P < .025 for ErbB3) and with corneal fluorescein staining (r = .409, P < .05 for ErbB2;r = .588, P < .005 for ErbB3). CONCLUSION: The expression of the type 1 growth factor receptors is significantly greater in the conjunctival epithelium of eyes with keratoconjunctivitis sicca than normal eyes. The increased expression of these receptors was positively correlated with ocular surface dye staining. The increased expression of these receptors may contribute to the abnormal growth and differentiation of the conjunctival

关键词：抗体单克隆印迹法蛋白质结膜/代谢结膜/病理学上皮/代谢上皮/病理学荧光抗体技术间接角结膜炎干燥性/代谢角结膜炎干燥性/病理学受体 erbB-2/生物合成受体 erbB-3/生物合成成年人老年人女(雌)性人类中年人

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Basic fibroblast growth factor synthesis by human peritoneal mesothelial cells - Induction by interleukin-1

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AMERICAN JOURNAL OF PATHOLOGY 1999年第6期155卷 1977-1984页

作者： Cronauer, MV Stadlmann, S Klocker, H Abendstein, B Eder, IE Rogatsch, H Zeimet, AG Marth, C Offner, FA Univ Innsbruck Dept Pathol A-6020 Innsbruck Austria Univ Innsbruck Dept Urol A-6020 Innsbruck Austria Univ Innsbruck Dept Obstet & Gynecol A-6020 Innsbruck Austria

Peritoneal mesothelial cells are uniquely located to regulate cellular events in the peritoneal cavity and are an important source for various cytokines and growth factors. This study was conducted to analyze the capacity of human peritoneal mesothelial cells (HPMCs) to synthesize and release basic fibroblast growth factor (bFGF) and to characterize its regulation by inflammatory cytokines. HPMCs constitutively synthesized and released considerable amounts of bFGF as detected by a specific immunoassay. Almost 80% of bFGF (1547 +/- 173 pg/10(5) cells) was localized intracellularly, Approximately 20% of the bFGF (357 +/- 27 pg/10(5) cells) was associated with extracellular matrix components on the HPMC surface. Small amounts of bFGF (<1%) were detectable in tissue culture supernatants (8.4 +/- 1.4 pg/10(5) cells). Treatment of HPMCs with interleukin-1 beta (IL-1 beta;1 ng/ml) resulted in a significant increase in bFGF production. The intracellular bFGF content showed a rapid but only transient increase, which was significant above background levels after 24 hours (41% increase;P < 0.05). This increase in intracellular bFGF concentration was associated with an induction of the release of bFGF, Within 96 hours, the release of bFGF to the cell surface and into the supernatant increased by 58% (564 +/- 52.4 pg/10(5) cells;P < 0.01) and by 214% (26.4 +/- 3.2 pg/10(5) cells;P < 0.001), respectively. Neither tumor necrosis factor-alpha nor interferon-gamma affected bFGF synthesis by HPMCs. Stimulation of HPMCs with IL-1 beta increased steady-state levels of bFGF-specific mRNA, Immunohistochemical analyses of peritoneal tissue revealed constitutive expression of bFGF by HPMCs. This in situ expression proved to be most pronounced in areas of serosal inflammation in activated HPMCs. Our study demonstrates that HPMCs synthesize and release significant amounts of bFGF and that the expression of this growth factor is significantly up-regulated by the proinflammatory cytokine IL-1

关键词：细胞培养的酶联免疫吸附测定上皮/代谢成纤维细胞生长因子2/生物合成成纤维细胞生长因子2/代谢免疫酶技术白细胞介素1/药理学白细胞介素1/生理学腹膜/细胞学腹膜/代谢 RNA 信使/分析逆转录聚合酶链反应肿瘤坏死因子α/药理学人类

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