黑斑蛙早期胚胎发育过程中乳酸脱氨酶同工酶的初步研究

本文采用聚丙烯酰胺凝胶电泳方法,对河南新乡地区黑班蛙（Rananigromaculata）早期胚胎发育过程中（受精后0－324h）乳酸脱氢酶（LDH）同工酶进行了研究。结果表明：LDH1自始至终存在,且活性一直占绝对优势;LDH5比于心跳期出现,该期以后其活性仅次于LDH1;LDH2于襄胚早期开始出现,其活性一直较弱;LDH3和LDH4均于开口期少量出现,以后前者活性一直极弱,后者则呈现一定的活性。与哺乳类及鱼类资料比较,提示两栖类黑斑蛙LDH同工酶有其独特的表达谱式。     

草鱼同工酶发育遗传学研究——Ⅱ.早期发育过程中的同工酶分析

采用淀粉凝胶电泳技术研究了草鱼早期发育过程中(受精后0—200小时)6种同工酶系统(LDH、MDH、GDH、ADH、IDH、EST)的表达谱式。除了ADH以外,其余5种同工酶系统均具有明显的发育变化谱式。根据早期发育过程中同工酶的变化谱式及其组织分布,草鱼的同工酶可分为三大类型:(1)在未受精卵及早期发育过程中一直存在,并常有较广泛的组织分布;(2)未受精卵及早期发育过程中均不存在,一般仅分布于少数几种组织中;(3)未受精卵及胚胎发育早期不存在,直到早期发育过程中某一特定时期才开始出现。     

异精效应在雌核发育彭泽鲫胚胎发育中的同工酶证据

采用聚丙烯酰胺凝胶垂直板电泳技术 ,对同源和异源精子激发雌核发育彭泽鲫子代胚胎发育过程中四种同工酶 (EST ,LDH ,MDH ,ME)的表达情况进行了比较研究。结果表明同源和异源精子激发的胚胎在MDH和LDH同工酶表达上存在明显差异。MDH同工酶的差异主要表现为 :在孵出期 ,异源精子激发的胚胎比同源精子激发的胚胎多出两条谱带MDH4’和MDH5’ ;LDH同工酶的差异表现为 :在原肠中期至肌肉收缩期的五个时期中 ,异源精子激发的胚胎比同源精子激发的胚胎多出 4条谱带 (LDH9’— 12’)。这种差异说明同源与异源精子对子代胚胎发育过程中同工酶表达的影响不同 ,可能属于“异精生物学效应”的一种表现形式     

青鱼胚胎发育中乳酸脱氢酶（LDH）同工酶的研究

本文报道了LDH同工酶基因在青鱼胚胎发育过程中表达的时序。研究结果表明,LDH-B基因在整过发育过程中部处于活动状态;胚胎发育早期的LDH同工酶谱与成熟卵的酶谱相同;LDH-A_2B_2、-A_3B和-A_4三种同工酶在胚胎发育至出膜前期后才出现。     

四类不同倍性鲫鱼在胚胎发育期间同工酶基因表达的比较分析

用聚丙烯酰胺梯度凝胶电泳比较分析了单倍体、二倍体、三倍体和复合四倍体4类不同倍性鲫鱼以及单倍体和二倍体鲤鱼在胚胎发育时期4种同工酶(EST,LDH,MDH,SOD)酶谱。结果表明,单倍体鲫鱼和单倍体鲤鱼胚胎与各自的二倍体胚胎相比,同工酶酶谱看不出差异;天然三倍体银鲫胚胎的MDH和SOD同工酶酶谱与二倍体鲫相似,但EST和LDH同工酶比二倍体增多了酶带,有的酶带如EST5和EST6还可在鲤鱼胚胎中找到相应的表达产物,提供了天然雌核发育三倍体银鲫杂交起源的证据;复合四倍体由于含有鲤鱼的一个外来基因组,其胚胎的基因表达有些与杂种类似,在所分析的4种同工酶酶谱中,都可观察到来自鲤鱼基因的影响。此外,在由源于不同复合四倍体个体的卵子发育形成的胚胎间,还观察到同工酶基因表达的异质性。     

鳙鱼同工酶发育遗传学研究

采用淀粉或聚丙烯酰胺凝胶电泳法分析鳙鱼早期发育阶段(从未受精卵到卵黄吸尽期)及成体不同组织(脑、眼、心、肌、肾、肝)中六种同工酶(LDH,MDH,IDH,ADH,SDH,EST)的分化表达模式。鳙鱼同工酶基因的表达具有明显的组织特异性。早期发育阶段,ADH和SDH均无染色活性;LDH、MDH和IDH具有不同的发育变化谱式,而EST酶谱在整个早期发育阶段均无明显变化。与鲢、草鱼相比,鳙鱼早期发育过程中胚胎Ldh-A基因激活的时间被推迟。上述结果可为鳙鱼种群的生化遗传结构分析以及鳙鱼的人工育种提供基础资料。     

兔单个植入前克隆胚胎cDNA文库的构建

人与小鼠和牛在正常胚胎植入前发育过程中基因活化的研究已经取得了长足的进展 ,但是还没有对同期克隆胚胎相关研究的报道 .利用单个家兔植入前移核重构胚胎成功地构建了MⅡ卵母细胞及发育至 4 、8 细胞期的胚胎和囊胚的特异性cDNA文库 .并用 β肌动蛋白和LAPTM4α证实这类文库是可靠的 .以 8 细胞期移核重构胚cDNA文库为例 ,随机挑取克隆进行测序分析 :其中 2 3的基因EST片段可以在GenBank或EST库中找到同源序列 ,约 1 3的EST片段属于未知的新片段 ,表明这是一类重要的新兴基因资源库 (期特异性EST库 ) .这种利用单胚胎构建cDNA文库的方法 ,解决了胚胎研究材料受限的问题 ,在时间上更加精确 ,更符合胚胎发育的规律 ,也能够更加准确地反映出一些克隆胚胎的异常表型 ,是研究早期胚胎发育基因表达以及克隆胚胎再程序化基因表达的一种有效手段 .     

5-脱氧杂氮胞苷抑制小鼠附植前的胚胎发育

DNA甲基化在哺乳动物发育过程中有关键作用．在小鼠附植前胚胎发育过程中,DNA甲基化一直处于动态变化过程中．通过将体外受精胚在5-AZA-CdR中持续培养,研究5-AZA-CdR对小鼠附植前胚胎发育的影响,为附植前胚胎发育机理的研究及5-AZA-CdR的毒副作用研究提供试验基础．从原核期加入不同浓度的5-AZA-CdR时,胚胎不能发育到桑椹胚(0.2 和1.0 μmol/L)和4-细胞胚(5.0 μmol/L);从2-细胞期加入时,胚胎阻滞于未致密化的8-细胞(0.2 和1.0 μmol/L)和3/4-细胞期(5.0 μmol/L);而当从4-细胞加入时,虽然胚胎能够发育到早期桑椹胚,但发育比例同对照相比显著降低(P < 0.05)．进一步检测凋亡、基因组DNA甲基化和整体转录活性,结果显示,高浓度的5-AZA-CdR导致8-细胞和早期桑椹胚发生早期凋亡,而低浓度的5-AZA-CdR引起8-细胞和早期桑椹胚基因组DNA甲基化的降低和转录活性的降低,并且这种降低呈浓度依赖性．所以加入低浓度的5-AZA-CdR时,胚胎的DNA甲基化降低,引起转录活性的降低,进而导致胚胎发育的停滞．     

蟾蜍种间核移植胚胎发育早期LDH同工酶的表现

利用聚丙烯酰胺凝胶电泳,对中华大蟾蜍(Bufo bufo比gargarizans)与花背蟾蜍(Bufo raddei)种间核移植胚胎发育早期六个不同阶段中全胚胎的乳酸脱氢酶(LDH)同工酶进行了分析。酶谱比较的结果表明:在正、反种间移核胚胎中,供体核LDH基因的活动开始表现于尾芽胚期;此前,杂种胚胎中LDH同工酶谱类型与受体一致。     

团头鲂的胚胎及成体组织中八种同工酶系统的研究

用垂直的淀粉凝胶电泳方法分析了胚胎发育阶段(0—105小时)和成体6种组织中的乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)、谷氨酸脱氢酶(GDH)、葡萄糖—6—磷酸脱氢酶(G6PD)、醇脱氢酶(ADH)、异柠檬酸脱氢酶(IDH)、酯酶(EST)和碱性磷酸酶(AKP)等8种同工酶系统的酶带。共约有23个基因座位在其胚胎发育期和成体组织中表达。所分析的大多数同工酶在团头鲂个体发生过程中表达的情况大致可分为3种类型：①胚胎发育期间持续存在的同工酶类,它们在成体组织中无特异性分布;②到胚胎发育后期才开始表达的同工酶类,它们往往和特定的组织或器官的形态发生或机能分化紧密相关,并在成体组织中呈特异性分布;③在成体组织中都没有被发现而只在胚胎发育期所特有的同工酶。团头鲂的MDH同工酶类之间以及它们和G6PD同工酶活性变化之间存在着相关性,二者可能存在共同的调控机制。与许多其他鱼类不同,团头鲂的GDH同工酶在整个胚胎发育过程中都有活性,但在其成体组织中无特异性分布。基因调控系统的时空精确性是保证团头鲂胚胎发育正常代谢活动的必要条件。     

Lactate dehydrogenase isoenzymes in loach (Misgurnus fossilis) development

Isoenzyme patterns of lactatedehydrogenase (LDH, EC 1.1.1.27) were determined in the loach (Misgurnus fossilis) oocytes, unfertilized eggs, developing embryos and larvae, and the sensitivity of the isoenzymes to the effect of AgNO3, p-chlormercurybenzoate and high temperature were investigated. The presence of 4 phenotypes of LDH patterns which differed in the isozymes number, in their relative activity, in electrophoretic mobility in the polyacrylamide gel columnes and in the frequency of occurence in different populations of the investigated species is demonstrated. Before the hatching stage there appeared a new isozyme which was completely inhibited by AgNO3 and had the same electrophoretic mobility in all LDH phenotypes. During the larvae development the content of this LDH isozyme increased. Some isozymes, especially those which had low anodic mobility and decreased in their relative quantity, disappeared during the development of embryos and larvae, p-Chlormercurybenzoate in the concentration 1 mM and heating at 70 degrees C during 20 minutes did not significantly affect LDH activities in the loach eggs, embryos and larvae.     

黄鳝性逆转过程中同工酶的分析研究 Ⅰ．雌雄个体不同组织的同工酶分析

作者采用三种垂直板聚丙烯酰胺凝胶电泳对雌雄黄鳝６种组织的乳酸脱氢酶（ＬＤＨ）、酯酶（ＥＳＴ）进行了研究。结果表明ＬＤＨ同工酶谱为细带型,可分为３区８条带,其迁移率为Ａ＞Ｂ＞Ｃ,其中Ｃ＿４仅出现在心、脑和肌肉中。ＬＤＨ在肝中表达甚微。ＥＳＴ同工酶显示３区７条带。黄鳝的同工酶有明显的组织特异性,在两性中的表达也存在一定的差异,这种差异ＥＳＴ比ＬＤＨ明显;肝、肾和性腺比其它组织较明显。这两种同工酶在雌性中的表达均比在雄性中强。以上反映了差别的基因活性。本文还讨论了黄鳝同工酶的遗传基础,亚基的组成以及ＬＤＨ迁移率和黄鳝演化地位的关系。     

菊花不同生长阶段不同器官POD和EST同工酶比较

采用过氧化物酶(POD)、酯酶(EST)2个酶系统的12个同工酶位点,分析了4个菊花品种营养生长和生殖生长阶段不同器官(嫩叶、老叶、嫩茎、木质化茎)的同工酶变化.结果表明:(1)4个品种共有16种POD酶带,15种EST酶带;(2)菊花的POD和EST具有组织特异性和阶段特异性,其中以嫩叶的酶带最多,其次为老叶,再次为嫩茎,而木质化茎的酶带最少;(3)与生殖生长阶段相比,营养生长阶段的POD酶带更清晰,更整齐,分离更好,但生殖生长阶段的EST同工酶比营养生长阶段的更清晰;(4)营养生长阶段的嫩叶最适合用于菊花POD同工酶分析,而EST同工酶研究宜取生殖生长时期的嫩叶.     

白鲢个体发育过程中同工酶基因的表达与调控研究

采用淀粉或聚丙烯酰胺凝胶电泳及特异性组织化学染色技术研究了白链早期发育过程中（从未受精卵到卵黄吸尽期）及成体不同组织（脑、眼、心、肌、肾、肝）中六种同工酶系统（ＬＤＨ,ＭＤＨ,ＩＤＨ,ＡＤＨ,ＳＤＨ,ＥＳＴ）的分化表达谱式,白链各同工酶基因的表达具有明显的组织特异性早期发育过程中,ＡＤＨ和ＳＤＨ基因均无活性,其它四种同工酶则具有不同的个体发育谱式。值得一提的是,由取自同一地区的两对白链的受精卵的形     

红螯螯虾胚胎发育期主要消化酶和同工酶的活性变化

采用生物化学方法测定了红螯螯虾(Cherax quadricarinatus)胚胎发育各期主要消化酶(胃蛋白酶、胰蛋白酶、淀粉酶、纤维素酶和脂肪酶)的比活力及主要同工酶(乳酸脱氢酶、苹果酸脱氢酶、葡萄糖-6-磷酸脱氢酶和酯酶)的活力。结果显示,5种消化酶各自表现出不同的变化模式,胃蛋白酶和胰蛋白酶的比活力早期均逐渐上升,到发育后期胃蛋白酶出现快速下降,而胰蛋白酶却仍保持较高水平;淀粉酶比活力呈“V”字型变化趋势,晚期活性较高;纤维素酶和脂肪酶的比活力则均较低。4种同工酶酶谱随胚胎的发育渐趋复杂,酶活性也随之增强。结果表明,消化酶和同工酶活力的高低均受其基因的调控,并随胚胎发育适时表达,为胚胎组织、器官和系统的形成以及未来仔虾的开口摄食提供物质保证。     

Postnatal development of the energy supplying enzyme pattern in the rat brain

The activity of seven enzymes connected with energy-supplying metabolism was followed from the second day of life till adulthood (87th day). The enzymes selected were: 1. Triosephosphate dehydrogenase (TPDH), 2. Lactate dehydrogenase (LDH), 3. Glycerol-3-phosphate: NAD dehydrogenase (GPDH), 4. Hexokinase (HK), 5. Malate: NAD dehydrogenase (MDH), 6. Citrate syntase (CS) and 7. 3-Hydroxyacyl Co A dehydrogenase. Although some variations occurred, the enzyme profiles were characteristic of those of the nervous tissue from the second day of life onwards until adulthood and displayed relatively high activities of HK, CS and MDH and low activities of TPDH, LDH, GPDH and HOADH. The activities of all enzymes studied here increased during postnatal development and some reached adult values on the 14th day, that of TPDH on the 27th day and HOADH on the 41st day of life. The activities of MDH and GPDH did not attain the adult values still on the 41st day of life. The anaerobic energy supply capacity seems to increase transiently on the 31st day of life, i.e. at a developmental stage where the resistance against hypoxia is known to increase transiently.     

Changes in myoglobin and lactate dehydrogenase in muscle tissues of a diving bird, the Pigeon Guillemot, during maturation

1. The concentration of myoglobin (Mb) and the isozymic distribution and activity of lactate dehydrogenase (LDH) in heart and pectoralis muscle were investigated at three stages of maturation of the Pigeon Guillemot, Cepphus columba. 2. Mb is not detectable in chick pectoralis; it is present in fledgling pectoralis muscle and increases four-fold in adult pectoralis. Mb concentration in heart muscle is similar in chick and fledgling and doubles in the adult. 3. LDH activities in pectoralis muscle of fledgling and adult increase to about three times that of the chick. LDH activities in heart of chick, fledgling and adult are similar to one another. 4. All five isozymes of LDH are present in both heart and pectoralis muscle at all stages; the heart muscle shows predominantly LDH-1 isozyme, and the pectoralis, LDH-5. The relative amounts of the five isozymes in the heart extract were constant during maturation but pectoralis LDH isozymes changed during maturation towards a more even distribution of the five isozymes in the adult. 5. Changes in Mb and LDH in the Pigeon Guillemot correlate with the animal's maturation from a sedentary nest sitter to an active diver and flyer. The adult pectoralis muscle probably has both aerobic function for wing-propelled short dives and flying and anaerobic capacity for longer dives.     

THE POSTNATAL DEVELOPMENT OF CATECHOL-O-METHYL TRANSFERASE IN THE RAT BRAIN

Abstract— The postnatal development of three enzymes in the rat forebrain was studied. When expressed per tissue weight the catechol- O -methyl transferase (COMT) increased 2-fold from birth to adult age, the lactate dehydrogenase (LDH) 4-fold and the monoamine oxidase (MAO) 12-fold. Expressed per mg protein the increase in the enzyme activities in the subcellular fractions which contained the main part of the different enzymes was still 2–4-fold for COMT and LDH while for MAO it was 4-fold.
There was a relative increase in the COMT activity in the P₂ fraction (synaptosomes and mitochondria). This increase was identical with a corresponding increase in LDH activity and protein and was probably due to growth of nerve terminals. The COMT in the cytoplasm of the synaptosomes showed the same increase relative to the proteins as did the 'free' cytoplasmic enzyme.
The conclusion is drawn that the enzymes in the rat brain show a certain degree of development during brain growth. An additional increase of some enzymes is due to the development of specialized structures such as mitochondria and nerve terminals with synapses. COMT is not related to any such specialized structure.     

香果树体细胞胚胎发生过程中4种同工酶的研究

用非变性聚丙烯凝胶电泳技术对香果树体细胞胚胎发生及形态建成过程中过氧化物酶(POD)、酯酶(EST)、淀粉酶(AMY)和超氧化物歧化酶(SOD)4种同工酶进行分析.结果表明:香果树体细胞胚胎发生及形态建成过程中,POD、EST、AMY和SOD活性变化与胚性愈伤组织的诱导及体细胞胚的发生发育密切相关.非胚性愈伤组织和胚性愈伤组织酶谱差异明显,胚性愈伤组织中EST和AMY同工酶酶带多且活性高,非胚性愈伤组织中缺乏EST和AMY同工酶表达,AMY同工酶可作为胚性细胞分化和发育的重要标志.香果树体细胞胚形态建成过程中,球形胚时期的AMY、POD、EST同_T酶活性最强,表明这一时期生理代谢旺盛,是体细胞胚形态建成的关键时期;POD、AMY和SOD 3种同工酶的酶谱及表达强弱在形态建成的不同时期呈现有规律的变化,可作为香果树体细胞胚发生发育特定时期的参考标记. 与胚性愈伤组织的诱导及体细胞胚的发生发育密切相关.非胚性愈伤组织和胚性愈伤组织酶谱差异明显,胚性愈伤组织中EST和AMY同工酶酶带多且活性高,非胚性愈伤组织中缺乏EST和AMY同工酶表达,AMY同工酶町作为胚性细胞分化和发育的重要标志.香果树体细胞胚形态建成过程 ,球形胚时期的AMY、POD、EST同_T酶活性最强,表明这一时期生理代谢旺盛,是体细胞胚形态建成的关键时期;POD、AMY和SOD 3种同工酶的酶谱及表达强弱在形态建成的不同时期呈现有规律的变化,可作为香果树体细胞胚发生发育特定时期的参考标记. 与胚性愈伤组织的诱导及体细胞胚的发生发育密切相关.非胚性愈伤组织和胚性愈伤组织