不同处理对动物线粒体DNA提取的影响

无脊推动物^[1]野桑蚕的蛹分别在不同温度下保存处理和用不同方法破碎新鲜蛹细胞处理，脊推动物^[1]长吻鮠的离体肝脏在不同试剂下保存处理。经过处理后的材料分别再用改良的碱变性法^[2-6]提取mtDNA，并比较相应处理的mtDNA。研究发现：这几种处理的材料都能够将其mtDNA提取出来，但不同处理的材料线粒体DNA质量有所不同，提取动物线粒体DNA时应根据实际需要处理材料。     

硒对镉中毒小鼠肝细胞线粒体超微结构影响的体视学研究

用电镜定性观察和体视学量定相结合的方法研究镉中毒小鼠肝细胞线粒体超微结构的变化及硒的保护作用。电镜定性观察发现，镉中毒小鼠肝细胞线粒体极度肿胀，嵴消失乃至空泡化。硒镉联合处理小鼠有也类似的变化，但程度较轻，体视学计量发现Ｃｄ处理小直细胞线粒体平均体积、体密度、平均外径面密度、平均截面积和平均截面周长显著高于硒镉联合处理１小鼠和正常小鼠，面数密度、比表面和数密度显著低于硒镉联合处理小鼠和对照小鼠；硒     

玉米赤霉烯酮对小鼠脾淋巴细胞凋亡的影响

采用MTT法、DNA ladder分析、流式细胞仪分析等方法,离体研究玉米赤霉烯酮对小鼠脾淋巴细胞增殖与凋亡的影响,结果发现:ZEA对离体培养LPS活化小鼠脾淋巴细胞增殖具有显著抑制作用(P<0.05),且抑制强度与其剂量和处理时间均呈依赖性关系;小鼠脾淋巴细胞经ZEA处理后,出现DNA断裂等典型细胞凋亡特征;ZEA对离体培养LPS活化小鼠脾淋巴细胞具有显著促凋亡作用(P<0.0 5),且促进强度与其剂量呈依赖性关系。这些结果表明,玉米赤霉烯酮对小鼠脾淋巴细胞有直接毒害作用。     

玉米赤霉烯酮对小鼠胸腺上皮细胞线粒体的影响

为探讨玉米赤霉烯酮对离体培养的小鼠胸腺上皮细胞的作用,本研究以1月龄昆明小鼠为研究对象,采用MTT法、线粒体染色、流式细胞仪分析等方法,离体研究玉米赤霉烯酮对胸腺上皮细胞线粒体的影响。结果表明,高剂量(10~25 μg/mL)玉米赤霉烯酮能显著增强小鼠胸腺上皮细胞线粒体代谢MTT能力,增强线粒体活性,使线粒体膜电位升高。     

玉米赤霉烯酮对小鼠脾淋巴细胞凋亡的影响

采用MTT法、DNA ladder分析、流式细胞仪分析等方法,离体研究玉米赤霉烯酮对小鼠脾淋巴细胞增殖与凋亡的影响,结果发现:ZEA对离体培养LPS活化小鼠脾淋巴细胞增殖具有显著抑制作用(P<0.05),且抑制强度与其剂量和处理时间均呈依赖性关系;小鼠脾淋巴细胞经ZEA处理后,出现DNA断裂等典型细胞凋亡特征;ZEA对离体培养LPS活化小鼠脾淋巴细胞具有显著促凋亡作用(P<0.0 5),且促进强度与其剂量呈依赖性关系.这些结果表明,玉米赤霉烯酮对小鼠脾淋巴细胞有直接毒害作用.     

家蚕蛹线粒体DNA EcoRⅠ 1.9kb片段的克隆及序列分析

克隆了二化性家蚕蛹体的线粒体DNAEcoRⅠ酶解 1 .9kb片段 ,并进行了序列测定。结果表明 :克隆片段中包含完整的线粒体赖氨酸转移RNA(tRNA Lys)基因、天冬氨酸转移RNA(tRNA Asp)基因、ATP合酶亚基Ⅷ基因和细胞色素C氧化酶亚基Ⅱ、ATP合酶亚基Ⅵ基因的部分序列。并与果蝇线粒体DNA的同源性和参照果蝇线粒体DNA密码子翻译的氨基酸顺序进行了比较 ,由tRNA的一级结构推断出可能的二级结构模型     

特定电磁波辐射离体猪精液对其解冻后精液品质及受精能力的影响

特定电磁波对家畜离体精液的处理效果,已在牛、绵羊、家兔等方面有报道。本研究使用ZH—3型TDP 辐射器,以32mw/cm~2(T_1组)、96mw/cm~2(T_3组)、160mw/cm~2(T_5)和224mw/cm~2(T_7组)辐射离体猪精液,在冷冻前和冷冻解冻后测定有关生理指标,包括透明质酸酶、顶体完整率、吖啶橙 DNA 荧光染色     

生物炭降低草莓根系线粒体膜功能伤害缓解乙草胺药害

为探究除草剂乙草胺对草莓根系线粒体膜功能及保护酶系统活性的影响,明确土壤中施加生物炭对减缓乙草胺对草莓根系线粒体功能伤害的保护机制。以盆栽草莓‘宝交早生’(Fragaria × ananassa Duch.)为试材,设置除草剂乙草胺处理、乙草胺+生物炭处理及对照3个处理,结果表明,乙草胺处理导致根系线粒体膜透性增大,根系线粒体膜电位Δψm下降,根系活力和H+ - ATPase含量降低,O2-、H2O2产生速率持续升高,保护性酶SOD、POD活性持续增长,MDA含量不断增加。乙草胺能够破坏草莓根系线粒体膜结构,增加细胞质膜过氧化程度,造成线粒体膜功能下降。施加生物炭能有效缓解乙草胺对线粒体膜功能的伤害,提高草莓植株对乙草胺的适应能力。     

猪卵母细胞线粒体分布及线粒体DNA拷贝数变化

本研究旨在观察猪卵母细胞线粒体分布及线粒体DNA拷贝数变化,以期作为判定哺乳动物卵母细胞胞质成熟的指标,同时也为今后克隆技术的发展和相关基因表达调控的研究提供基础.运用线粒体分子探针标记技术检测体外成熟不同时期卵母细胞中线粒体的分布变化,运用实时荧光定量PCR技术检测其线粒体DNA拷贝数的变化趋势,揭示线粒体分布、线粒体DNA拷贝数变化与卵母细胞发育潜能的关系.结果表明,猪卵母细胞成熟前后,线粒体分布由未成熟的周边分布变为成熟后的均匀分布,并且线粒体簇变大,着色变深.卵母细胞成熟0、11、22 h的mtDNA拷贝数分别为(2 519.52士940.39)、(3 421.47士345.71)和(9 747.58士1 928.24),他们之间无显著性差异(P＞0.05).卵母细胞成熟33 h的mtDNA拷贝数为(39 913.61±1 180.26),显著高于成熟0、11和22 h的mtDNA拷贝数(P＜0.05).卵母细胞成熟44 h的mtDNA拷贝数为(130 074.30±78 119.45),显著高于成熟33 h的mtDNA拷贝数(P＜0.05).由此可见,随着卵母细胞成熟进程的推进,线粒体活性增强,线粒体DNA拷贝数明显增加.     

线粒体DNA及蜜蜂线粒体DNA的研究现状与展望

线粒体DNA是真核细胞中核外唯一的遗传物质,是目前研究起源进化及群体遗传分析的理想研究对象。综述了线粒体DNA(mtDNA)基本特征、多态性及其分析方法的同时,介绍了蜜蜂线粒体DNA的组成结构和多态性,阐述了蜜蜂线粒体DNA研究的现状并对其未来的发展前景予以展望。     

线粒体DNA(mtDNA)的研究进展

本文概述了mtDNA的基本特征和研究方法；介绍了人及各种畜禽mtDNA的研究现状，并对mtDNA的研究作了展望。     

线粒体DNA及其在家畜遗传育种中的应用

对线粒体DNA的数目、大小、基本结构、多态性及其研究方法、遗传特性等方面进行了论述,并进一步阐述了线粒体DNA的多态性在家畜遗传育种中的应用现状。     

Cytotoxic Effects of Serum from Equine Grass Sickness Cases on Neuro-2a and PC12 Tet-Off Cell Lines: Implication for Using In Vitro Methods as Antemortem Diagnostic Tools

A wide variety of clinical and paraclinical methods have been used for diagnosis of equine grass sickness (EGS), but none of them could absolutely confirm the diagnosis, and postmortem pathologic examination is still considered the final step in precise diagnosis of EGS. Use of in vitro cell toxicity caused by EGS serum on neuronal cell lines was investigated. Three well-known cytotoxic methods were used to investigate the cytotoxicity of EGS serum on neuro-2a and genetically engineered PC12 Tet-Off P53 cells. The results of alamar blue reduction as an index for mitochondrial activities and intracellular adenosine triphosphate content assays, but not neutral red uptake, indicate that the EGS serum may affect the mitochondrial function and cellular metabolism at up to 60% of the cases. The results of present study might be used for diagnosis of EGS cases. Further studies with high sample size may lead us to uncover the pathogenesis of EGS and to increase the sensitivity and applicability of in vitro techniques as diagnostic tools.     

线粒体DNA与山羊进化

线粒体DNA由于其母系遗传、高拷贝、高进化速率以及缺乏重组等特点而被广泛的应用于山羊的起源进化研究.本文就山羊的多母系起源、弱的地理结构、群体扩张及分化时间等方面的mtDNA研究概况作了说明,并对线粒体DNA数据分析方法作了简要的介绍.     

线粒体DNA与山羊进化

线粒体DNA由于其母系遗传、高拷贝、高进化速率以及缺乏重组等特点而被广泛的应用于山羊的起源进化研究。本文就山羊的多母系起源、弱的地理结构、群体扩张及分化时间等方面的mtDNA研究概况作了说明,并对线粒体DNA数据分析方法作了简要的介绍。     

基于pcox1基因分析黑斑蛙小吻对盲囊线虫种系发育关系

以湖南省不同地区黑斑蛙体内采集的16条小吻对盲囊线虫样品为研究对象,利用PCR对其线粒体DNA中的细胞色素c氧化酶第1亚基(cox 1)基因部分序列(pcox 1)进行扩增,分析其遗传变异情况。并利用软件Clustal X 2.0对其序列进行分析,探讨小吻对盲囊线虫与其他线虫的种群遗传关系。测序结果显示,所获得的pcox 1序列长度一致,均为400 bp,湖南省各分离株之间相应序列的相似性在97%以上,与GenBank中登录的其他小吻对盲囊线虫pcox 1序列的相似度均低于89%。结果表明,小吻对盲囊线虫pcox 1序列种内相对保守,种间差异较大,可以作为种间遗传变异研究的分子标记,从而为小吻对盲囊线虫的分子流行病学调查研究等奠定基础。     

Comparison of two methods of seminal plasma removal on buffalo (Bubalus bubalis) sperm cryopreservation

Cryopreservation causes damage to spermatozoa, and methods minimizing this damage are therefore needed. Although much discussed, seminal plasma removal has become an alternative to improve sperm quality and viability after freezing and has been applied to different species in attempt to obtain good results. The objective of this study was to evaluate semen quality in buffaloes submitted to two methods for seminal plasma removal (filtration and centrifugation). Semen samples were collected from seven Murrah buffalo bulls (Bubalus bubalis) once a week for 8 weeks. Each ejaculate was divided into three groups: control (presence of seminal plasma), centrifugation and filtration. Sperm kinetics was evaluated with the computer‐assisted sperm analysis (CASA) system. Plasmalemma and acrosomal membrane integrity, mitochondrial membrane potential and reactive oxygen species (ROS) were measured by flow cytometry, and lipid peroxidation was evaluated by the thiobarbituric acid reactive substances (TBARS) assay. Seminal plasma removal did not improve sperm kinetics compared to the control group. Centrifugation increased the number of cells with damaged acrosomal membranes (0.77 ± 0.05) and filtration caused greater plasmalemma and acrosomal membrane damage (22.18 ± 1.07). No difference in the mitochondrial membrane potential was observed between groups. In contrast, ROS production was higher in the centrifugation group compared to the control and filtration groups, although no differences in TBARS formation were detected. In conclusion, seminal plasma removal did not improve the quality of thawed buffalo semen compared to control in terms of sperm kinetics, membrane integrity, mitochondrial membrane potential or lipid peroxidation.     

Identification of mitochondrial DNA substitutions related to meat quality in Japanese Black cattle

Complete sequences of mitochondrial (mt) genomes of eight Japanese Black cattle were determined to investigate the relationships between mt deoxyribonucleic acid (DNA) displacement loop (D-loop) types and other mtDNA regions and to identify the variation in the coding region that may influence the economic traits. The survey of mitochondrial sequences in the encoding region revealed 14 substitutions including six antonymous substitutions and one in 16S ribosomal ribonucleic acid (rRNA). Three methods of polymorphic DNA analyses (polymerase chain reaction [PCR]-restriction fragment length polymorphism [RFLP], mismatch PCR-RFLP, PCR-single-strand conformation polymorphism [SSCP]) were performed on these seven candidate substitutions (base pair [bp] 2,232, 12,158, 12,908, 13,310, 14,122, 14,140, and 14,565) for 202 Japanese Black cattle. The substitution of bp 13,310 was observed in all samples, but not in the reference sequence, indicating that this is a minor substitution or a sequencing mistake in the reference sequence. The substitutions at bp 14,122, 14,140, and 14,565 were observed in only a few samples, suggesting that these were also minor substitutions. The substitutions at bp 2,232 (16S rRNA), 12,158, and 12,908 (reduced nicotinamide adenine dinucleotide-ubiquinone oxidoreductase chain-5) were closely related to mitochondrial D-loop types that have previously been related to differences in the carcass traits of Japanese Black cattle. Evaluation of the effects on six carcass traits with mixed model procedures suggests that the bp 2,232 substitution affects longissimus muscle area and beef marbling score. The substitution at bp 2,232 is a strong candidate for the mitochondrial effect on meat quality.     

家蚕（Bombyx mori）线粒体DNA的经济快速提取方法

对碱裂解法进行了改良,采用不同pH值的均浆缓冲液从家蚕5龄幼虫后部丝腺．蛹和卵中提取mtDNA,调查其得率和纯度,建立了家蚕线粒体DNA经济快速提取方法。