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1.
Scaffold attachment regions stimulate HSP70.1 expression in mouse preimplantation embryos but not in differentiated tissues. 总被引:4,自引:1,他引:3 下载免费PDF全文
E M Thompson E Christians M G Stinnakre J P Renard 《Molecular and cellular biology》1994,14(7):4694-4703
Eukaryotic interphase chromatin is thought to be organized into topologically discrete, independent domains acting as units upon which differential patterns of gene expression are established. Sequences which attach chromatin to in vitro preparations of a nucleoprotein matrix (scaffold attachment regions [SARs]) may act as domain boundaries, but their role remains poorly defined compared with those of other elements such as locus control regions. We have produced mice homozygous for a transgene which is transcribed as early as the activation of the embryonic genome at the two-cell stage and which is expressed ubiquitously in a number of differentiated tissues. Transgenic lines were generated in the presence or absence of flanking SAR sequences, creating an original model which enabled us to examine the effects of these elements at different developmental stages. In the preimplantation mouse embryo, flanking SARs stimulated transgene expression in a copy-dependent manner. In contrast, in the differentiated tissues of newborn and adult mice, no significant SAR-dependent increase in transgene expression was found, correlation with copy number was lost, and position effects were observed. These results suggest a limited capacity of SARs to act as insulating elements but are consistent with a proposed model of SAR-mediated chromatin opening and closing. 相似文献
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Beta-adrenergic induced potassium current in Xenopus oocyte: involvement of cyclic AMP 总被引:1,自引:0,他引:1
The effect of a beta-adrenergic agonist, on full-grown Xenopus oocytes, still surrounded by their ovarian envelopes, has been studied by electrophysiological methods. The oocytes were hyperpolarized by isoproterenol. Under voltage clamp, the elicited outward current reversed at a membrane potential of - 95 mV, a value close to the K+ equilibrium potential. The isoproterenol induced current varied linearly with the membrane potential in the range studied (- 120 mV, - 30 mV). Half-maximum current was obtained at 3.10(-8) M isoproterenol. Propranolol (10(-7) M) completely suppressed the response to isoproterenol (10(-9) to 10(-5) M). 8-Br-cAMP induced a current which also reversed at - 95 mV. Methyl-isobutyl-xanthine (MIX), a potent inhibitor of phosphodiesterases, potentiated the current induced by isoproterenol. These experiments strongly suggest that the increase in K+ permeability due to catecholamines is mediated by cAMP. 相似文献
3.
Ouanounou G Malo M Stinnakre J Kreger AS Molgo J 《The Journal of biological chemistry》2002,277(42):39119-39127
Trachynilysin, a protein toxin isolated from the venom of the stonefish Synanceia trachynis, has been reported to elicit massive acetylcholine release from motor nerve endings of isolated neuromuscular preparations and to increase both cytosolic Ca2+ and catecholamine release from chromaffin cells. In the present study, we used the patch clamp technique to investigate the effect of trachynilysin on the cytoplasmic membrane of differentiated NG108-15 cells in culture. Trachynilysin increased membrane conductance the most when the negativity of the cell holding membrane potential was reduced. The trachynilysin-induced current was carried by cations and reversed at about -3 mV in standard physiological solutions, which led to strong membrane depolarization and Ca2+ influx. La3+ blocked the trachynilysin current in a dose-, voltage-, and time-dependent manner, and antibodies raised against the toxin antagonized its effect on the cell membrane. The inside-out configuration of the patch clamp technique allowed the recording of single channel activity from which various multiples of 22 pS elementary conductance were resolved. These results indicate that trachynilysin forms pores in the NG108-15 cell membrane, and they advance our understanding of the toxin's mode of action on motor nerve endings and neurosecretory cells. 相似文献
4.
Langley Brett Vilotte Jean-Luc Stinnakre Marie-Georges Whitelaw C. Bruce A. L'Huillier Phil J. 《Transgenic research》1998,7(3):205-212
In an attempt to enhance the frequency and level of expression of a poor-performing MMTV-driven transgene, we co-integrated this construct with the ovine -lactoglobulin (BLG) gene in transgenic mice. Seven lines of transgenic mice possessing co-integrated BLG and MMTV-RZ5 transgenes were compared with 12 lines of mice that possessed only the MMTV-RZ5 construct. Co-integration enhanced the frequency of expression in the mammary gland from two out of 12 lines for the MMTV-RZ5 transgene alone, to five out of seven when co-integrated with BLG. Surprisingly, co-integration also resulted in co-expression of the two transgenes in the salivary gland, lung and spleen in addition to the mammary gland. Furthermore, both transgenes were expressed in virgin animals, and throughout pregnancy and lactation, suggesting that the developmental regulation of the locus follows that of the MMTV-promoter. These findings represent a novel locus control property of the ovine BLG gene that confer s commitment of the locus to the mammary gland, but also to a range of heterogeneous tissues possibly defined by the second promoter at the locus 相似文献
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The external and internal (lumen) diameter and the height of the epithelium of the Wolffian ducts of control rat fetuses were measured between 14 days 8 hrs and 21 days 8 hrs at the level of the gonad and in the genital folds. In males, at the level of the gonad, these ducts display a transitory increase in diameter and lumen at 16 days 8 hrs of fetal age. This increase, which is absent in females, occurs just before the development of the epididymides and might reflect the endocrine activity of the fetal testis. In the females, the first involutive changes appear at both levels at 17 days 8 hrs of age (decrease in diameter by reduction of the lumen and height of the epithelium). After injections of androgens to pregnant rats or directly into the fetuses in utero, the Wolffian ducts can be maintained in female fetuses only if submitted to androgens before and until 16 days 16 hrs. If the treatment starts once the involutive changes have appeared (17 days 8 hrs) inconstant persistence is obtained. The portions of ducts still present on day 18 cannot be maintained by androgens any more. Even if injected at 15 days 8 hrs, exogenous androgens do not hasten or anticipate the formation of Wolfian derivatives (epididymides and seminal vesicles) in males or in females. 相似文献
9.
Eve Devinoy Marie-George Stinnakre Franoise Lavialle Dominique Thpot Michle Ollivier-Bousquet 《Experimental cell research》1995,221(2)
Growth hormone (GH) secretion, in mammary tissue from transgenic mice, containing a chimeric gene composed of the regulatory region of whey acidic protein gene and the structural region of GH gene, was compared to casein secretion. GH was expressed in milk and for a small percentage (1:1000) in blood as revealed by SDS-polyacrylamide gel electrophoresis and radio-immunoassay. As attested by immunofluorescence and immunogold electron microscopy, caseins and GH followed the same secretory pathway. However, contrary to caseins, which are essentially in micellar form, GH was detected in a nonaggregated form in secretory vesicles and in the lumen of the acini. Newly synthesized caseins and GH were carried simultaneously, mainly to the lumen of the acini, but also to the base of the cell. Secretion of newly synthesized proteins was increased by prolactin (PRL). As shown by immunoblotting, the proportion of GH versus other proteins, secreted in the presence of PRL was not modified, suggesting that GH secretion is subjected to the same hormonal regulation by PRL as other milk proteins. These results show that, in lactating mammary epithelial cells from transgenic mice, a recombinant GH and the caseins are carried simultaneously to the lumen and suggest that secretion of both proteins is increased by PRL during the same time course. Transport of these newly synthesized proteins occurs also to the base of the cell. 相似文献
10.
Markedly increased susceptibility to natural sheep scrapie of transgenic mice expressing ovine prp 下载免费PDF全文
Vilotte JL Soulier S Essalmani R Stinnakre MG Vaiman D Lepourry L Da Silva JC Besnard N Dawson M Buschmann A Groschup M Petit S Madelaine MF Rakatobe S Le Dur A Vilette D Laude H 《Journal of virology》2001,75(13):5977-5984
The susceptibility of sheep to scrapie is known to involve, as a major determinant, the nature of the prion protein (PrP) allele, with the VRQ allele conferring the highest susceptibility to the disease. Transgenic mice expressing in their brains three different ovine PrP(VRQ)-encoding transgenes under an endogenous PrP-deficient genetic background were established. Nine transgenic (tgOv) lines were selected and challenged with two scrapie field isolates derived from VRQ-homozygous affected sheep. All inoculated mice developed neurological signs associated with a transmissible spongiform encephalopathy (TSE) disease and accumulated a protease-resistant form of PrP (PrPres) in their brains. The incubation duration appeared to be inversely related to the PrP steady-state level in the brain, irrespective of the transgene construct. The survival time for animals from the line expressing the highest level of PrP was reduced by at least 1 year compared to those of two groups of conventional mice. With one isolate, the duration of incubation was as short as 2 months, which is comparable to that observed for the rodent TSE models with the briefest survival times. No survival time reduction was observed upon subpassaging of either isolate, suggesting no need for adaptation of the agent to its new host. Overexpression of the transgene was found not to be required for transmission to be accelerated compared to that observed with wild-type mice. Conversely, transgenic mice overexpressing murine PrP were found to be less susceptible than tgOv lines expressing ovine PrP at physiological levels. These data argue that ovine PrP(VRQ) provided a better substrate for sheep prion replication than did mouse PrP. Altogether, these tgOv mice could be an improved model for experimental studies on natural sheep scrapie. 相似文献