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61.
Isolation of ouabain-resistant human diploid fibroblasts 总被引:17,自引:0,他引:17
Seventeen clones resistant to the cytotoxic action of ouabain were isolated in culture by direct selection from 5 independent strains of diploid human fibroblasts. Resistant clones were recovered at frequencies on the order of 10?7 per wild type cell selected from populations treated with the mutagen EMS, but no resistant cells were detected among 108 unmutagenized cells. Most selected clones remained ouabain-resistant following further propagation in the absence of drug. The growth of wild type cells was inhibited by 50% at ouabain concentrations of 2–5 × 10?8 M, while resistant clones required 15–180 fold higher drug concentrations to cause equivalent inhibition. Ouabain-resistant clones showed increased resistance of K+ transport function to ouabain inhibition that paralleled their increased resistance to growth inhibition. Initial experiments suggest that under selective conditions the resistant diploid fibroblasts differ significantly from wild type in binding of 3H-ouabain per unit surface area. The ouabain-resistant cells were similar to wild type in transport properties unrelated to ouabain inhibition. Resistant cells had normal karyotypes and senesced with a lifespan similar to control clones. The ouabain-resistant phenotypes of these diploid human fibroblast isolates apparently reflect point mutations that specifically affect the Na+/K+ transport ATPase with respect to ouabain-binding and/or response to bound ouabain. 相似文献
62.
本文报道了中国产的黑腹菌属Melanogaster(黑腹菌科 Melanogastraceae,黑腹菌目 Melanogastrales,担子菌亚门Basidiomycotina)二新种:山西黑腹菌Melanogaster shanxicnsis Liu,K.Tao et Chang和倒卵孢黑腹菌M.obouatisporus Liu,K.Tao et Chang并对中国已报道过的黑腹菌属内的种进行了讨论。 相似文献
63.
Sadis Matalon Joseph S. Beckman Michael E. Duffey Bruce A. Freeman 《Free radical biology & medicine》1989,6(6):557-564
The purpose of this study was to quantify the effects of extracellularly generated partially reduced oxygen species on active sodium (NA+) transport across the ventral toad skin, a well-studied epithelium. Sections of skin from decapitated toads were mounted in an Ussing chamber, bathed on both sides with electrolyte solution containing 500 μM xanthine and bubbled continuously with room air. The tissues were short-circuited, and short circuit current (Isc) and tissue resistance (Rt were monitored continuously with an automatic voltage clamp apparatus. Fifteen mU/ml of xanthine oxidase (XO), either purchased from Calbiochem or purified from cream, were instilled in either the apical (mucosal) or basolateral (serosal) baths at t = 0 and T = 10 min. Hydrogen peroxide (H2O2) concentrations increased to 200 μM within the first 20 min and then decreased, reaching a value of 40 μM by 60 min. Mean [H2O2] was 90 μM. Instillation of XO in the apical bath resulted in a large decrease in Isc and an increase in Rt, their values being 43% and 160% of their corresponding controls 85 min after the first instillation. Addition of superoxide dismutase and catalase completely prevented these changes. Instillation of XO in the basolateral bath had no effect. Similar physiological responses were obtained using the Calbiochem XO or the purified XO, which contained no measurable protease activity. It was concluded that extracellularly generated partially reduced oxygen species may interfere with active Na+ transport by possibly damaging apical Na+ channel proteins. 相似文献
64.
Decreased Protein Kinase C Activity During Cerebral Ischemia and After Reperfusion in the Adult Rat 总被引:6,自引:4,他引:2
R. Christian Crumrine George Dubyak Joseph C. LaManna† 《Journal of neurochemistry》1990,55(6):2001-2007
The possible activation of protein kinase C (PKC) during total cerebral ischemia was investigated in the rat. Translocation of PKC activity from the soluble to the particulate fraction was used as an index of PKC activation. There was a drop in the proportion of particulate PKC activity from 30% for controls to 20% by 30 min of ischemia (p less than 0.01). By 20 min of cardiac arrest, there was a 40% decline of the total cellular PKC activity (p less than 0.01). This was not accompanied by an increase in activator-independent activity, a finding indicating PKC was not being converted to protein kinase M. These data suggest that PKC was not activated during ischemia, but rather that ischemia causes a reduction in cellular PKC activity. Translocation of PKC activity to the particulate fraction was not observed in the cerebral cortex or hippocampus of reperfused brain for up to 6 h of recovery following 11-13 min of total cerebral ischemia. The level of total, soluble, and particulate PKC activity in the cerebral cortex was reduced (p less than 0.05), corresponding to the decrease observed by 15 min of ischemia without reflow. A similar decline in activity was also observed in the hippocampus. No increase in activator-independent activity was observed. These data suggest that PKC was inhibited during cerebral ischemia and that this reduced level of PKC activity was maintained throughout 6 h of recovery. We conclude that pathological activation of PKC was not responsible for the evolution of ischemic brain damage. 相似文献
65.
Summary Isolated nerve cells fromLymnaea stagnalis were studied using the internal-perfusion and patch-clamp techniques. Patch excision frequently activated a voltage-independent Ba2+-permeable channel with a slope conductance of 27 pS at negative potentials (50mm Ba2+). This channel is not seen in patches on healthy cells and, unlike the voltage-dependent Ca channel, is not labile in isolated patches. The activity of the channel in inside-out patches is unaffected by intracellular ATP, Ca2+ below 1mm or the catalytic subunit of cAMP-dependent protein kinase but is reversibly blocked by millimolar intracellular Ca2+ or Ba2+. The channel can be activated in on-cell patches by either internal perfusion with high Ca2+ or the long-term internal perfusion of low Ca2+ solutions not containing ATP. These channels may carry the inward Ca2+ current which causes a regenerative increase in intracellular Ca+ when snail neurons are perfused with high Ca2+ solutions. High internal Ca2+, or long periods of internal perfusion with ATP-free solutions, induces an increase in a resting (–50 mV) whole-cell Ba2+ conductance. This conductance can be turned off by returning the intracellular perfusate to a low Ca2+ solution containing ATP and Mg2+. The activity of this channel appears to have an opposite dependence on intracellular conditions to that of the voltage-dependent Ca channel. 相似文献
66.
Lu Changya David V. Gallacher Robin F. Irvine Barry V. L. Potter Ole H. Petersen 《The Journal of membrane biology》1989,109(1):85-93
Summary We have examined the effects of various inositol polyphosphates, alone and in combination, on the Ca2+-activated K+ current in internally perfused, single mouse lacrimal acinar cells. We used the patch-clamp technique for whole-cell current recording with a set-up allowing exchange of the pipette solution during individual experiments so that control and test periods could be directly compared in individual cells. Inositol 1,4,5-trisphosphate (Ins 1,4,5 P3) (10–100 m) evoked a transient increase in the Ca2+-sensitive K+ current that was independent of the presence of Ca2+ in the external solution. The transient nature of the Ins 1,4,5 P3 effect was not due to rapid metabolic breakdown, as similar responses were obtained in the presence of 5mm 2,3-diphosphoglyceric acid, that blocks the hydrolysis of Ins 1,4,5 P3, as well as with the stable analoguedl-inositol 1,4,5-trisphosphorothioate (Ins 1,4,5 P(S)3) (100 m). Ins 1,3,4 P3 (50 m) had no effect, whereas 50 m Ins 2,4,5 P3 evoked responses similar to those obtained by 10 m Ins 1,4,5 P3. A sustained increase in Ca2+-dependent K+ current was only observed when inositol 1,3,4,5-tetrakisphosphate (Ins 1,3,4,5 P4) (10 m) was added to the Ins 1,4,5 P3 (10 m)-containing solution and this effect could be terminated by removal of external Ca2+. The effect of Ins 1,3,4,5 P4 was specifically dependent on the presence of Ins 1,4,5 P3 as it was not found when 10 m concentrations of Ins 1,3,4 P3 or Ins 2,4,5 P3 were used. Ins 2,4,5 P3 (but not Ins 1,3,4 P3) at the higher concentration of 50 m did, however, support the Ins 1,3,4,5 P4-evoked sustained current activation. Ins 1,3,4 P3 could not evoke sustained responses in combination with Ins 1,4,5 P3 excluding the possibility that the action of Ins 1,3,4,5 P4 could be mediated by its breakdown product Ins 1,3,4 P3. Ins 1,3,4,5 P4 also evoked a sustained response when added to an Ins 1,4,5 P(S)3-containing solution. Ins 1,3,4,5,6 P5 (50 m) did not evoke any effect when administered on top of Ins 1,4,5 P3. In the absence of external Ca2+, addition of Ins 1,3,4,5 P4 to an Ins 1,4,5 P3-containing internal solution evoked a second transient K+ current activation. Readmitting external Ca2+ in the continued presence internally of Ins 1,4,5 P3 and Ins 1,3,4,5 P4 made the response reappear. We conclude that both Ins 1,4,5 P3 and Ins 1,3,4,5 P4 play crucial and specific roles in controlling intracellular Ca2+ homeostasis. 相似文献
67.
This study was conducted to identify lines of subterranean clover (Trifolium spp.) with resistance to Meloidogyne arenaria (Neal, 1989) Chitwood, 1949, race 1; M. incognita (Kofoid and White, 1919) Chitwood, 1949, race 3; and M. javanica (Treub, 1885) Chitwood, 1949. A collection of 134 subterranean clover lines was evaluated and all had intermediate to high susceptibility. Root galling was negatively correlated with both seed and dry matter yields. Soil fumigation significantly reduced the nematode population in the field. Results indicate there is limited genetic resistance to root-knot nematodes among subterranean clover lines. Alternative sources of variation for this trait should be investigated. 相似文献
68.
新疆准噶尔盆地三个泉地区几种始新世哺乳类 总被引:8,自引:5,他引:3
本文记述了准噶尔盆地北缘三个泉地区依希白拉组中采到的踝节类、蹠行类、全齿类和恐角类化石。根据这些化石,依希白拉组的时代可能和北美中始新世勃力吉期相当。文内,通过文献分析,认为周明镇和胡长康(1956)记述的真恐角兽的前臼齿可能产自三个泉附近的依希白拉组。 相似文献
69.
Christiane Gulat-Marnay rée Lafitte Jean-Michel Arrang Jean-Charles Schwartz 《Journal of neurochemistry》1989,52(1):248-254
The cholinergic modulation of histamine release and synthesis was studied in rat brain slices or synaptosomes labeled with L-[3H]histidine. Carbachol in increasing concentrations progressively reduced the K+-induced [3H]histamine release from cortical slices. Pirenzepine, a preferential M1-receptor antagonist, reversed the carbachol effect in an apparently competitive manner and with Ki values of 1-6 X 10(-8) M. 11-[(2-[(Diethylamino)methyl]-1-piperidinyl)acetyl]-5,11-dihydro-6H- pyrido[2,3-b][1,4]benzodiazepine-6-one (AF-DX 116), considered a preferential M2-receptor antagonist, reversed the carbachol effect with a mean Ki of approximately 2 X 10(-7) M. Oxotremorine behaved as a partial agonist in the modulation of histamine release. Neostigmine, an acetylcholinesterase inhibitor, inhibited the K+-induced release of [3H]histamine from cortical slices, and the effect was largely reversed by pirenzepine, an observation suggesting a modulation by endogenous acetylcholine. The effects of carbachol and pirenzepine were observed with slices of other brain regions known to contain histaminergic nerve terminals or perikarya, as well as with cortical synaptosomes. The two drugs also modified, in opposite directions, [3H]histamine formation in depolarized cortical slices. In vivo oxotremorine inhibited [3H]histamine formation in cerebral cortex, and this effect was reversed by scopolamine. When administered alone, scopolamine failed to enhance significantly the 3H- labeled amine formation, a finding suggesting that muscarinic receptors are not activated by endogenous acetylcholine released under basal conditions. It is concluded that muscarinic heteroreceptors, directly located on histaminergic nerve terminals, control release and synthesis of histamine in the brain. These receptors apparently belong to the broad M1-receptor category and may correspond to a receptor subclass displaying a rather high affinity for AF-DX 116. 相似文献
70.