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1.
The relationship was studied between radiation-induced apoptosis in the nervous system of Drosophila larvae and the age dynamics in adult fly neuromuscular activity. The level of apoptosis in the neural ganglia of third-instar larvae from the wild-type strain increased 2.5 times after larval exposure to ionizing radiation (54 cGy). Irradiation of the strain with enhanced sensitivity to apoptosis induction, which carries a mutation in gene-inhibitor of apoptosis th (allele th4), and the wild-type strain Berlin led to an increase in neuromuscular activity of adult flies throughout the experiment and, consequently, to reduced aging rate. Conversely, this effect was not observed in strains with reduced sensitivity to induction of apoptosis (with mutations in genes dArk and Dcp-1).  相似文献   

2.
《Fly》2013,7(1):7-15
General anesthetics achieve behavioral unresponsiveness via a mechanism that is incompletely understood. The study of genetic model systems such as the fruit fly Drosophila melanogaster is crucial to advancing our understanding of how anesthetic drugs render animals unresponsive. Previous studies have shown that wild-type control strains differ significantly in their sensitivity to general anesthetics, which potentially introduces confounding factors for comparing genetic mutations placed on these wild-type backgrounds. Here, we examined a variety of behavioral and electrophysiological endpoints in Drosophila, in both adult and larval animals. We characterized these endpoints in 3 commonly used fly strains: wild-type Canton Special (CS), and 2 commonly used white-eyed strains, isoCJ1 and w1118. We found that CS and isoCJ1 show remarkably similar sensitivity to isoflurane across a variety of behavioral and electrophysiological endpoints. In contrast, w1118 is resistant to isoflurane compared to the other 2 strains at both the adult and larval stages. This resistance is however not reflected at the level of neurotransmitter release at the larval neuromuscular junction (NMJ). This suggests that the w1118 strain harbors another mutation that produces isoflurane resistance, by acting on an arousal pathway that is most likely preserved between larval and adult brains. This mutation probably also affects sleep, as marked differences between isoCJ1 and w1118 have also recently been found for behavioral responsiveness and sleep intensity measures.  相似文献   

3.
The SHIRPA protocol was proposed as a rapid, comprehensive screening method for qualitatively abnormal phenotypes in the mouse (Rogers et al., Mamm Genome 8, 711, 1997). This screening technique is currently being used to identify mutants induced by N-ethylnitrosourea (ENU) mutagenesis (Brown and Nolan, Hum Mol Genet 7, 1627, 1998). SHIRPA can be used to identify mutants with neuromuscular abnormalities, but the sensitivity of the protocol is unknown. We tested two dystrophin-deficient mutants Dmd mdx and Dmd mdx3cv , both of which are indistinguishable from wild-type by a simple visual assessment, at different ages, using the primary screen of the SHIRPA protocol. The most dramatic observation was that both Dmd mdx and Dmd mdx3cv mice showed extreme fatigue after testing, while mice from the same C57BL strains appeared unaffected. Each strain of dystrophin-deficient mice showed a different profile in locomotor activity and deficiencies in the wire maneuver, righting reflex, and negative geotaxis tests. Furthermore, the wire maneuver test indicated an earlier onset of muscular impairment in Dmd mdx than Dmd mdx3cv mice. These data suggest that the SHIRPA primary screen is effective not only in identifying subtle neuromuscular mutants, but also in distinguishing qualitative differences between mutants with neuromuscular abnormalities. Received: 5 August 1999 / Accepted: 14 April 2000  相似文献   

4.
曾成城  陈锦平  魏虹  刘媛  马文超  王婷  周翠 《生态学报》2017,37(13):4327-4334
三峡库区消落带面临水淹及Cd污染双重胁迫,为探究秋华柳(Salix variegata Franch.)在水淹条件下对Cd污染土壤的修复能力,以秋华柳扦插苗为试验材料,设置正常供水(CK)和水淹组(FL)两个水分处理方式,4个Cd浓度梯度:对照组(0mg/kg)、低浓度(0.5mg/kg)、中浓度(2mg/kg)及高浓度(10mg/kg),分别对处理60 d和120 d的土壤微生物数量及酶活性变化特征进行研究。试验结果表明:(1)Cd浓度处理均未显著影响土壤微生物数量(P0.05),水淹显著降低处理60 d土壤细菌数、真菌数及处理120 d的土壤放线菌数及真菌数(P0.05)。(2)种植秋华柳显著提高处理60 d土壤细菌数量(P0.05),对土壤放线菌、真菌数量也有一定提升。(3)Cd浓度处理显著影响处理60 d土壤磷酸酶活性及处理120 d脲酶活性(P0.05),水淹显著降低处理60 d土壤磷酸酶活性及处理120 d脲酶、蔗糖酶和磷酸酶活性(P0.05)。(4)正常供水及水淹条件下,种植秋华柳对土壤酶活均有一定改善作用。种植秋华柳显著提高了处理60 d土壤磷酸酶活性以及处理120 d脲酶和蔗糖酶活性(P0.05)。研究结果表明:水淹生境中,秋华柳对Cd污染土壤微生物数量及酶活性具有改善作用,在Cd污染土壤修复方面有一定应用前景。  相似文献   

5.
Cytochrome P450 17α-hydroxylase/17, 20 lyase (CYP17) is a microsomal enzyme reported to have two distinct catalytic activities, 17α-hydroxylase and 17, 20 lyase, that are essential for the biosynthesis of peripheral androgens such as dehydroepiandrosterone (DHEA). Paradoxically, DHEA is present and plays a role in learning and memory in the adult rodent brain, while CYP17 activity and protein are undetectable. To determine if CYP17 is required for DHEA formation and function in the adult rodent brain, we generated CYP17 chimeric mice that had reduced circulating testosterone levels. There were no detectable differences in cognitive spatial learning between CYP17 chimeric and wild-type mice. In addition, while CYP17 mRNA levels were reduced in CYP17 chimeric compared to wild-type mouse brain, the levels of brain DHEA levels were comparable. To determine if adult brain DHEA is formed by an alternative Fe2+-dependent pathway, brain microsomes were isolated from wild-type and CYP17 chimeric mice and treated with FeSO4. Fe2+ caused comparable levels of DHEA production by both wild-type and CYP17 chimeric mouse brain microsomes; DHEA production was not reduced by a CYP17 inhibitor. Taken together these in vivo studies suggest that in the adult mouse brain DHEA is formed via a Fe2+-sensitive CYP17-independent pathway.  相似文献   

6.
【目的】铜绿假单胞菌(Pseudomonas aerugionsa)二鸟苷酸环化酶SiaD调控着铜绿假单胞菌的生物被膜形成等表型。在研究过表达siaD对生物被膜的调控作用时发现,与野生型siaD基因回补菌株相比,一株回补菌株的生物被膜产量显著升高。本文的目的即是探究该菌株生物被膜产量升高的原因,并对该菌株的其他表型进行研究。【方法】通过测序确定突变位点;利用生物被膜定性和定量实验对发生点突变的菌株表型进行分析;利用Western blotting实验检测SiaDR119M蛋白表达水平;利用GST-pulldown实验检测SiaC蛋白与SiaDR119M蛋白在体外的结合能力;针对siaDR119M点突变基因进行融合蛋白表达载体的构建,表达并纯化该蛋白,利用高效液相色谱检测SiaDR119M的酶活;为了进一步研究c-di-GMP与细菌运动能力的关系,对细菌的运动能力进行检测。【结果】测序比对结果显示,序列的第119个氨基酸发生了突变,由精氨酸突变成了甲硫氨酸。生物被膜定性和定量实验显示,与野生型siaD...  相似文献   

7.
Summary Growth, citric acid production and enzymatic activity of the mitochondrial respiratory enzymes of a wild-type and a citric-acid-producing mutant of Aspergillus niger have been compared during fermentation under citric-acid-accumulating and non-accumulating conditions. Under non-accumulating conditions, both strains showed standard growth and no citric acid production. The mutant strain was characterized by delayed onset of growth and lowered cell yield. Under citric-acid-accumulating conditions the wild-type strain exhibited decelerated growth and a maximal citric acid concentration of 12 g l–1. Reduced, but continuing growth and citric acid production of 32 g l–1 was observed for the mutant strain. In general, the mutant strain exhibited reduced activity for the proton-pumping respiratory complexes and enhanced activity for the alternative respiratory enzymes. In contrast to the stable activity of complex I in the wild-type strain, this complex was selectively lost in the mutant strain at the onset of citric acid production, while the alternative NADH dehydrogenases were kept at enhanced and constant activity. A possible causal connection between the loss of complex I and citric acid accumulation is discussed. Offsprint requests to: J. Wallrath  相似文献   

8.
General anesthetics achieve behavioral unresponsiveness via a mechanism that is incompletely understood. The study of genetic model systems such as the fruit fly Drosophila melanogaster is crucial to advancing our understanding of how anesthetic drugs render animals unresponsive. Previous studies have shown that wild-type control strains differ significantly in their sensitivity to general anesthetics, which potentially introduces confounding factors for comparing genetic mutations placed on these wild-type backgrounds. Here, we examined a variety of behavioral and electrophysiological endpoints in Drosophila, in both adult and larval animals. We characterized these endpoints in 3 commonly used fly strains: wild-type Canton Special (CS), and 2 commonly used white-eyed strains, isoCJ1 and w1118. We found that CS and isoCJ1 show remarkably similar sensitivity to isoflurane across a variety of behavioral and electrophysiological endpoints. In contrast, w1118 is resistant to isoflurane compared to the other 2 strains at both the adult and larval stages. This resistance is however not reflected at the level of neurotransmitter release at the larval neuromuscular junction (NMJ). This suggests that the w1118 strain harbors another mutation that produces isoflurane resistance, by acting on an arousal pathway that is most likely preserved between larval and adult brains. This mutation probably also affects sleep, as marked differences between isoCJ1 and w1118 have also recently been found for behavioral responsiveness and sleep intensity measures.  相似文献   

9.
The biochemical mechanisms for growth tolerance to a 100% CO headspace in cultures, and butanol plus ethanol production from CO by Butyribacterium methylotrophicum were assessed in the wild-type and CO-adapted strains. The CO-adapted strain grew on glucose or CO under a 100% CO headspace, whereas, the growth of the wild-type strain was severely inhibited by 100% CO. The CO-adapted strain, unlike the wild-type, also produced butyrate, from either pyruvate or CO. The CO-adapted strain was a metabolic mutant having higher levels of ferredoxin–NAD oxidoreductase activity, which was not inhibited by NADH. Consequently, only the CO-adapted strain can grow on CO because CO oxidation generates reduced ferredoxin which, via the mutated ferredoxin–NAD reductase activity, forms reduced NADH required for catabolism. When the CO-adapted strain was grown at pH 6.0 it produced butanol (0.33 g/l) and ethanol (0.5 g/l) from CO and the cells contained the following NAD-linked enzyme activities (μmol min−1 mg protein−1): butyraldehyde dehydrogenase (227), butanol dehydrogenase (686), acetaldehyde dehydrogenase (82) and ethanol dehydrogenase (129). Received: 15 September 1998 / Received revision: 12 February 1999 / Accepted: 19 February 1999  相似文献   

10.
We propose a model to describe the changes taking place in biochemical processes/events to explain the development of heterocyst and nitrogenase in a diazotrophic cyanobacterium Anabaena variabilis. For this purpose, a mutant strain of A. variabilis lacking heterocyst differentiation and incapable of growth with dinitrogen as the sole source of nitrogen has been isolated after nitrosoguanidine (NTG) mutagenesis and selection by penicillin enrichment. The mutant strain (Het Fix) thus isolated has morphological variation and was incapable of reducing acetylene under anaerobic conditions, indicating its mutational loss of the process of nitrogen fixation. The Het Fix mutant strain had reduced glutamine synthetase (transferase) activity compared with its wild-type counterpart, suggesting a link between nif gene expression and the expression of gln A, the structural gene of GS. The Het Fix mutant strain compared with its wild-type strain also had an extremely high level of phycobiliprotein and a low level of carotenoids. Furthermore, the coiling of vegetative filaments in the Het Fix mutant strain, which reduced the surface area to be exposed to light, was a direct indication of the chromatic adaptation, because the mutant strain was found to be photosensitive, showing bleaching of the cells under high light intensity. Received: 13 December 2000 / Accepted: 9 February 2001  相似文献   

11.
Dihydrofolate reductase and aminopterin resistance in Pneumococcus   总被引:1,自引:0,他引:1  
Summary Wild-type pneumococci derived from Avery's strain R36A are sensitive to extracellular concentrations of the folate antimetabolite aminopterin exceeding 1.0x10-6 M. Three classes of resistant strains are phenotypically distinguishable: amiB-r, amiA-r and amiD-r strains are resistant to low (1.5x10-6 M), intermediate (0.5–4.0×10-5 M) and high (4.5x10-4 M) aminopterin levels respectively. The amiA and amiB regions are weakly linked, but linkage has not been established between either of these loci and the amiD region.Consistent with the maximum resistance conferred by mutations in the amiA locus, dihydrofolate (FH2) reductase in cell-free extracts (CFE) of amiA-r strains has a two- to six-fold greater affinity for the substrate than dose the enzyme in wild-type CFE (Table 1); FH2 reductase from amiA-r strains may also have reduced affinity for aminopterin. Specific activity of the enzyme is not affected by mutation in the amiA locus (Table 1) and its affinity for the cofactor (NADPH) is probably unaffected by mutation in this locus (Table 4). Dihydrofolate reductase activity in amiA5 CFE is considerably more thermolabile than that in wild-type CFE (Table 2).The enzyme in CFE of the high resistance strain amiD1 has the same affinity for the substrate, cofactor and antimetabolite as FH2 reductase in wild-type CFE (Figs. 1–4, 8 and 9; Table 4). However, specific activity of the enzyme in amiD1 CFE is 11-fold higher than that in wild-type CFE (Table 1) and it is much more heat stable (Table 2).Some properties of FH2 reductase in CFE of the high resistance recombinant strain amiA5amiD1 are intermediate between those in CFE of wild-type and amiD1.Preliminary results suggest that CFE of wild-type and amiA5 contain a factor, which is neither dialyzable nor heat sensitive, that has an inhibitory effect upon activity and stability of FH2 reductase in amiD1 CFE (Tables 2 and 3).  相似文献   

12.
Presenilins are one of two types of critical genetic factors in familial Alzheimer's disease, and they regulate various cellular functions such as intracellular Ca2+ homeostasis, the endoplasmic reticulum (ER) stress response, apoptosis, and synaptic transmission. We utilized Drosophila presenilin (psn) mutants as a model for studying the role of this gene in regulating acetylcholinesterase activity (AChE) and synaptic plasticity. Several lines of biochemical evidence indicated that AChE activity in a functionally null psn mutant (psnB3) was significantly reduced. In addition, we also found that psnB3 mutant neuromuscular junctions (NMJs) had smaller synaptic boutons and altered localization of Discs large, a synaptic scaffolding protein at the synaptic terminals compared to wild-type controls. These phenotypic defects were completely rescued in transgenic lines expressing the long form of wild-type Psn under an endogenous psn promoter cassette (PEPC-PsnWT;psnB3 lines). Taken together, these results indicate that Psn is important for regulating AChE activity, the size of synaptic boutons, and the localization of DLG at synaptic terminals.  相似文献   

13.
Properties of Mitomycin C-sensitive Mutants of Escherichia coli K-12   总被引:7,自引:4,他引:3  
Strains hypersensitive to mitomycin C (MC) were isolated from Escherichia coli K-12 after treatment with nitrosoguanidine. Of 43 MC-sensitive strains tested for their ultraviolet light (UV) sensitivity and for their ability to reactivate UV-inactivated λ phage, 38 were found to be insensitive to UV irradiation and to be able to reactivate UV-irradiated bacteriophage λ. Some properties of the MC-sensitive, uvr+ mutants were analyzed. Synthesis of deoxyribonucleic acid (DNA) in MC-sensitive, uvr+ mutants was inhibited at a lower concentration of MC than in the wild-type strain. Mutant cells, labeled with 3H-thymidine and then exposed to MC, released radioactivity as low molecular weight compounds. The amount of radioactivity released was the same as that from the wild-type strain. MC-sensitive, uvr+ mutants, as well as the corresponding wild-type strain, were equally susceptible to induction of prophage 80 by UV irradiation. However, MC induction of prophage was achieved in MC-sensitive, uvr+ mutants at a lower concentration of the antibiotic than in the wild-type strain. Genetic experiments indicated that a gene controlling MC sensitivity is located close to that determining lactose fermentation of E. coli. It is situated on episome F′13, and the wild type is dominant to the MC-sensitive allele.  相似文献   

14.
Temperature had various effects on the predacious efficacy of immature and mature stages of the coccinellid predator, Stethorus punctillum on the two-spotted spider mite, Tetranychus urticae. In the case of immature stages, food consumption at the lowest tested temperature (15°C) was significantly higher than that at higher temperatures (25 and 35°C). On the contrary, positive correlation between food consumption and temperature was evaluated in the case of adult predator. Regarding predator responses to different prey density, a high positive correlation between food consumption and prey density was evaluated among 4th instar larvae of the predator, followed by adult predator, while younger instars did not show reasonable increases with increasing prey densities. These results confirm that larval and adult stages of S. punctillum exhibit “Type II” functional response. In conclusion, the 4th instar larvae and adult predator are the most preferable stages in winter and summer crops to control T. urticae, respectively.  相似文献   

15.
Mutant strains sensitive and resistant to the drug 5-fluorouracil (FU) have been isolated from the wild-type Pac strain of Drosophila melanogaster. The resistant strain, termed flur, is resistant to at least 0.0035%FU (2.7 × 10–4 m) in the food media and exhibits cross-resistance to 5-fluorodeoxyuridine (FUdR) but not to 5-fluorouridine (FUR). The sensitive strain termed flu S , exhibits over 90% mortality on 0.0008% FU (6 × 10–5 m). Genetic analysis indicates that the flu gene is located on the third chromosome, which agrees with results of previous workers. An analysis of the enzyme thymidylate synthetase from the selected sensitive and resistant strains indicates that the resistant strain enzyme possesses an elevated specific activity. Levels 4 times that of the sensitive strain were observed when the enzymes were assayed at 20 C. This increase is apparently not due to induction by FU in the food media. It is suggested that the enzyme thymidylate synthetase may be involved in the resistance process.  相似文献   

16.
Allelic complementation between arg-7 mutants in Chlamydomonas reinhardi   总被引:1,自引:0,他引:1  
R. Loppes  R. Matagne 《Genetica》1972,43(3):422-430
In Chlamydomonas reinhardi, the arg-7 locus is the structural gene for the enzyme argininosuccinate lyase (ASL) which catalyzes the last reaction in arginine biosynthesisOut of the nine ASL--mutants so far investigated, seven were shown to fall within three distinct groups of complementation.The activity of ASL produced in diploids formed by complementation is much lower than the activity of ASL in the wild-type strain.The study of the heat sensitivity of the enzymes seems to indicate that in every diploid only one enzymic variety is formed which in most cases displays a much greater sensitivity to heat than the enzyme formed by the wild-type strain.These results are discussed in relation to the hypothesis according to which the arg-7 gene in Chlamydomonas reinhardi corresponds to one cistron and the ASL enzyme is composed of at least two identical polypeptide chains.Chercheur qualifié du Fonds National Belge de la Recherche Scientifique  相似文献   

17.
We investigated effects of vitamin C and E (VCE) administration on desflurane-induced oxidative toxicity and element changes in the blood of operative patients under desflurane general anesthesia. Forty American Society of Anesthesiologists I or II Physical Status adult patients were scheduled for elective surgery. The patients were randomly divided into two groups. Control and VCE group was introduced to anesthesia with desflurane. VCE was administreted to patients in the control and VCE group before 1 hour of anesthesia with desflurane. Baseline (preoperative) and postoperative (at the 1st, the 24th, and 72th h), blood samples were taken from the first and second groups. Erythrocyte and plasma lipid peroxidation levels at the 1st, 24th, and 72th hours were higher in the control than in baseline group, although their levels at the same periods were lower in the VCE group than in the control. Vitamin E levels at the postoperative 1st and 24th hours and erythrocyte glutathione peroxidase (GSH-Px) activity at the postoperative 1st, 24th, and 72th hours was lower than in baseline values. Erythrocyte GSH-Px activity and plasma vitamins A, C, and E levels at the postoperative 1st, 24th, and 72th hours were higher in the VCE group than in the control group. Erythrocyte and plasma reduced glutathione, plasma β-carotene, and serum copper, while zinc, selenium, aluminum, iron, magnesium, and calcium levels did not differ between preoperative and postoperative periods in both groups. In conclusion, VCE combination prevented the desflurane-induced vitamin E and GSH-Px consumptions to strengthen the antioxidant levels in the blood of operative patients.  相似文献   

18.
Chronic irradiation (accumulated dose 0.6–0.8 Gy) was shown to change the life span in male Drosophila melanogaster.Death was retarded in wild-type strains and accelerated in mutant strains defective in DNA repair and displaying a higher sensitivity to induction of apoptosis.  相似文献   

19.
Summary Yeast strain 990 carries a mutation mapping to the oli1 locus of the mitochondrial genome, the gene encoding ATPase subunit 9. DNA sequence analysis indicated a substitution of valine for alanine at residue 22 of the protein. The strain failed to grow on nonfermentable carbon sources such as glycerol at low temperature (20°C). At 28°C the strain grew on nonfermentable carbon sources and was resistant to the antibiotic oligomycin. ATPase activity in mitochondria isolated from 990 was reduced relative to the wild-type strain from which it was derived, but the residual activity was oligomycin resistant. Subunit 9 (the DCCD-binding proteolipid) from the mutant strain exhibited reduced mobility in SDS-polyacrylamide gels relative to the wild-type proteolipid. Ten revertant strains of 990 were analyzed. All restored the ability to grow on glycerol at 20°C. Mitotic segregation data showed that eight of the ten revertants were attributable to mitochondrial genetic events and two were caused by nuclear events since they appeared to be recessive nuclear suppressors. These nuclear mutations retained partial resistance to oligomycin and did not alter the electrophoretic behavior of subunit 9 or any other ATPase subunit. When mitochondrial DNA from each of the revertant strains was hybridized with an oligonucleotide probe covering the oli1 mutation, seven of the mitochondrial revertants were found to be true revertants and one a second mutation at the site of the original 990 mutation. The oli1 gene from this strain contained a substitution of glycine for valine at residue 22. The proteolipid isolated from this strain had increased electrophoretic mobility relative to the wild-type proteolipid.Abbreviations DCCD dicyclohexylcarbodiimide - SDS sodium dodecyl sulfate - PMSF phenylmethylsulfonyl fluoride - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonate - SMP submitochondrial particles - mit- mitochondrial point mutant  相似文献   

20.
Abstract

Nitric oxide is known to be a messenger in animals and plants. Catalase may regulate the concentration of intracellular ?NO. In this study, yeast Saccharomyces cerevisiae cells were treated with 1–20 mM S-nitrosoglutathione (GSNO), a nitric oxide donor, which decreased yeast survival in a concentration-dependent manner. In the wild-type strain (YPH250), 20 mM GSNO reduced survival by 32%. The strain defective in peroxisomal catalase behaved like the wild-type strain, while a mutant defective in cytosolic catalase showed 10% lower survival. Surprisingly, survival of the double catalase mutant was significantly higher than that of the other strains used. Incubation of yeast with GSNO increased the activities of both superoxide dismutase (SOD) and catalase. Pre-incubation with cycloheximide prevented the activation of catalase, but not SOD. The concentrations of oxidized glutathione increased in the wild-type strain, as well as in the mutants defective in peroxisomal catalase and an acatalasaemic strain; it failed to do this in the mutant defective in cytosolic catalase. The activity of aconitase was reduced after GSNO treatment in all strains studied, except for the mutant defective in peroxisomal catalase. The content of protein carbonyls and activities of glutathione reductase and S-nitrosoglutathione reductase were unchanged following GSNO treatment. The increase in catalase activity due to incubation with GSNO was not found in a strain defective in Yap1p, a master regulator of yeast adaptive response to oxidative stress. The obtained data demonstrate that exposure of yeast cells to the ?NO-donor S-nitrosoglutathione induced mild oxidative/nitrosative stress and Yap1p may co-ordinate the up-regulation of antioxidant enzymes under these conditions.  相似文献   

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