河蚌外套膜组织培养细胞分泌的珍珠质的药理作用

本文研究河蚌外套膜组织培养细胞分泌珍珠质的药理作用。组织培养后的培养液,能缩短小白鼠出血时间。用组织培养后的培养液及组织块水解液,对大鼠离体子宫及蟾蜍离体心脏收缩有增强作用,对兔离体小肠有抑制作用。这些药理作用与培养液中牛磺酸含量变化一致。实验结果表明,河蚌分泌珍珠质的细胞,在离体人工培养条件下,能旺盛地分泌珍珠质,分泌的珍珠质具有天然珍珠相同的一些药理作用。     

三角帆蚌外套膜及珍珠囊的组织学初步观察

近些年来,人们对海水马氏珠母贝(Pinctada martensii)和淡水池蝶蚌(Hyriopsis schlegelii)等双壳类外套膜及珍珠囊进行了组织学、组织化学和体外培养等的研究,然而结合河蚌人工育珠方面的报道却不多。事实上,欲提高淡水珍珠质量虽取决于多种因素,但其中与取供体蚌中的外套膜外表皮制作植片用的小片部位,以及珍珠囊的形态结构有着非常密切的关系。因此,本文旨在能对提高河蚌人工育珠的质量提供一些参考。     

三种不同因子对三角帆蚌外套膜细胞Ca~(2+)流动性的影响

<正>三角帆蚌(Hyriopsis cumingii)是我国主要的育珠母蚌,其所培育出的珍珠色泽鲜艳、细腻光滑,因而成为目前淡水育珠生产中首选的育珠母蚌之一。蚌的外套膜是贝壳和珍珠形成的重要组织器官,有内外两层表皮细胞及其间的结缔组织构成。它对钙具有高度通透性,其上皮细胞具有通过细胞膜主动吸收Ca~(2+)和贮存Ca~(2+)的功能,并通过胞吐作用排出钙至外套膜外腔中,这些钙就是形成珍珠的基础。维     

几种淡水蚌外套膜蛋白质氨基酸组成及部分元素分析

测定了三角帆蚌、圆背角无齿蚌、褶纹冠蚌、背视丽蚌外套膜的粗蛋白含量 ,分析了外套膜干粉的氨基酸组成 ,并采用原子吸收光谱法和等离子吸收光谱法测定三角帆蚌、圆背角无齿蚌、褶纹冠蚌外套膜的 12种元素含量。结果表明 :干燥外套膜蛋白质含量为 :三角帆蚌 4 1 34% ,圆背角无齿蚌2 2 0 3% ,褶纹冠蚌 32 14 % ,背视丽蚌 7 78%。这四种淡水蚌外套膜均含有 17种氨基酸。被测的 12种元素中 ,Cu元素含量最高 ,其次是Fe ,Mn ,Mg ,Ca ,Zn ,Cr,而Ni,Co ,Pb ,Hg等元素含量较低。这四种淡水蚌外套膜是一类优质的动物蛋白质资源。     

圆背角无齿蚌离体培养的外套膜组织钙代谢

本次实验采用离体组织培养技术研究外套膜组织的钙代谢,它排除了蚌体内的其他因素,如神经、激素等对外套膜生理、生化等方面的影响,并用组化方法对外套膜中钙及有关粘多糖的分布进行了观察研究,期望能进一步了解外套膜组织钙代谢调控的机理,同时,为淡水珍珠养殖业提供一些参考资料。       

褶纹冠蚌外套膜组织培养的分泌物的偏光显微镜观察

以淡水育珠贝中珍珠形成较快的褶纹冠蚌为材料,用相差显微镜观察组织培养的外套膜的分泌物的形成和变化,用偏光显微镜观察分泌物的双折射现象,并与活体外套膜的分泌物、贝壳的角质层、棱柱层、珍珠层的双折射现象进行比较。结果表明;离体培养的外套膜细胞不仅能产生活体细胞相同的分泌物,而且分泌物还能在培养过程中形成结晶,并逐渐生长。发现外套膜的不同部位分区培养所形成的分泌物的性状与结晶性质和活体有一致性,表明组织培养的外套膜小片具有贝体原来的组织结构、分化特征和分泌功能。     

三角帆蚌钙网蛋白基因cDNA的全长克隆与表达分析

采用同源克隆策略和RACE技术,从三角帆蚌（Hyriopsis cumingii）外套膜组织中成功克隆得到钙网蛋白（Calreticulin,CRT）基因的全长cDNA序列,共1838 bp,开放阅读框为1257 bp,编码418个氨基酸,5'端非编码区为75 bp,3'端非编码区为506 bp,基因序列提交GenBank的登录号为JX416227。生物信息学分析表明,三角帆蚌钙网蛋白基因具有一段信号肽序列、两条典型的钙网蛋白家族标签序列KHEQNIDCGGGY和IMFGPDICG、三个保守的N-、P-和C-端功能域及内质网前导序列HDEL。NJ法系统进化分析显示三角帆蚌首先与海洋双壳类紧密聚在一起,且与蚯蚓等环节动物亲缘关系较近,聚为一支,然后依次与虾类、昆虫、鱼类、两栖类、哺乳类聚在一起。经荧光定量PCR检测,钙网蛋白基因在三角帆蚌的外套膜、闭壳肌、斧足、鳃、肝脏、性腺、心脏、肠等8个组织中均有表达,其中在外套膜、鳃和斧足等与贝类钙代谢相关的组织中表达量较高预示其可能参与三角帆蚌的钙代谢。不同Ca2+浓度处理试验的结果表明,随着水体中Ca2+浓度逐渐升高,三角帆蚌钙网蛋白基因在外套膜中的表达水平呈先上升后下降的趋势,并在60 mg/L时达 到最高峰,表明适宜的Ca2+浓度可促进钙网蛋白基因表达,而过高的Ca2+浓度则会抑制其表达。同时在60 mg/L Ca2+浓度条件下,对三角帆蚌外套膜进行不同时间的表达试验,结果表明钙网蛋白基因的表达量随时间推移先上升,并于48h达到最大表达量,而后逐渐下降。上述结果为进一步深入研究钙网蛋白基因的功能及其调控机理奠定基础。       

褶纹冠蚌钩介幼虫非寄生变态发育观察及早期稚蚌的生长

为揭示褶纹冠蚌钩介幼虫变态发育特征及过程,采用体外培养方法实现了褶纹冠蚌钩介幼虫的非寄生变态发育。运用光学显微镜和扫描电子显微镜对变态发育过程中幼虫外部形态、内部器官发育进行了系列观察,对非寄生变态发育的稚蚌后期生长发育进行跟踪研究,并分析了底泥和光照两种环境因子对稚蚌存活及生长的影响。结果显示:在整个培养过程中,钩介幼虫的外部形态及大小未出现显著性变化,而斧足、鳃丝、外套膜及内脏团等组织器官逐步形成;在培养第3天,幼虫可见斧足雏形,鳃丝、外套膜纤毛尚未发现;在培养第6天,斧足成形,可见斧足侧沟,外套膜纤毛稀疏,鳃丝出现;培养第9天,斧足纤毛、外套膜纤毛增多,鳃丝密集。稚蚌投喂30d后,鳃丝基本成形。养殖试验结果表明:底泥对稚蚌存活和生长具有显著影响（P < 0.01）,而光照无显著性影响（P>0.05）。该结果为蚌科钩介幼虫变态发育生物学研究积累了基础资料,也通过对稚蚌生长的评估证实了体外培养是蚌类人工繁育及保护的有效技术途径。     

褶纹冠蚌外套膜组织培养的分泌物的偏光显微镜…

以淡水育珠贝中珍珠形成较快的褶纹冠蚌为材料,用相差显微镜观察组织培养的外套膜的分泌物的形成和变化,用偏光显微镜观察分泌物的双折射现象,并与活体外套膜的分泌物、贝壳的角质层、棱柱层、珍珠层的双折射现象进行比较。结果表明：离体培养的外套膜细胞不仅能产生活体细胞相同的分泌物,而且分泌物还能在培养过程中形成结晶,并逐渐生长。发现外套膜的不同部位分区培养所形成的分泌物的性状与结晶性质和活体有一致性,表明组织     

背角无齿蚌外套膜的组织化学研究

背角无齿蚌(Anodonta woodiana elliptica)在我国分布很广,是我国三种淡水育珠蚌之一。它所形成的珍珠微带金黄色,具有特殊的风格。四川省安岳县自1971年开始使用背角无齿蚌进行珍珠养殖以来,获得了不少经济利益,并积累了一些育珠经验。为了进一步探讨珍珠形成机理,以利于提高珍珠质量,从而获得更多优质珍珠。我们在对我国淡水育珠河蚌外套膜组织学和染色体等研究的基础上,又采用组织化学方法,对背角无齿蚌外套膜细胞中的核酸、糖类、酶等生物大分子物质作了组织化学方面的一些观察研究。     

淡水育珠蚌体外培养外套膜细胞分泌的珍珠质的性质研究

对淡水育珠蚌分泌珍珠质的外表皮进行组织培养,取培养不同时间的培养基,用氨基酸分析仪对其氨基酸测定的结果与空白培养基作比较,其中珍珠所含的各种氨基酸均增加,尤其是珍珠中含量最高的丙氨酸,甘氨酸和谷氨酸增加最多,珍珠药用有效成分之一的牛磺酸,随组织培养时间的延长而逐渐增加,用等离子光谱测定培养组织匀浆液与未培养比较,钙的含量也大为增加,结果表明,离体组织培养分泌的珍珠质的化学成分和性质与活体基本相同。     

Taurine-Induced Regeneration of Goldfish Retina in Culture May Involve a Calcium-Mediated Mechanism

Abstract: A possible mechanism of action of taurine as a trophic substance was studied in goldfish retina by investigating the effect of extracellular and intracellular calcium chelators on in vitro outgrowth, and the effect of taurine on calcium influx into postcrush retinal cells in culture. The amino acid stimulated the outgrowth from goldfish retinal explants, an effect that was blocked by EGTA and 1,2-bis ( o aminophenoxy)ethane- N,N,N',N' -tetraacetic acid methyl ester (BAPTA). The influx of calcium into cultured cells from postcrush retina was increased by taurine by day 5 in culture, but not by day 10, supporting previous results indicating a critical period in which taurine stimulates out-growth. The present results suggest that taurine partially exerts its regenerative effect on postcrush retinal explants by increasing calcium influx.     

培养条件对三角帆蚌外套膜离体上皮组织珍珠质分泌能力的影响（简报）

珍珠是由珍珠贝外套膜的上皮细胞受到外源刺激物刺激形成珍珠囊（pearl—sac）,并由珍珠囊产生的钙质分泌物．其分泌物逐渐包围刺激原．使之体积急剧增长而形成的,珍珠质（nacre）是由大于95％的碳酸钙晶体与约5％的角壳蛋白组成的生物矿化产物。因此珍珠贝的外套膜在珍珠形成中起着重要的作用。珍珠贝外套膜的体外培养已开展了一些初步的研究工作。     

牛磺酸对乳鼠心肌细胞内钙浓度的影响

研究低氧、复氧对乳鼠心肌细胞内钙离子浓度的影响,以及牛磺酸在模拟心肌缺血/再灌注(I/R)过程中对细胞内钙的调节作用。采用SD大鼠乳鼠进行心肌细胞培养,建立模拟I/R模型。以Fluo-4/AM荧光指示剂负载,应用激光共聚焦显微镜技术(confocal laser scanning microscope,CLSM)检测心肌细胞钙离子浓度的变化。对照组心肌细胞内钙离子荧光强度(23.71±2.37U)较低;低氧180 min后复氧即刻,钙离子荧光强度开始增加(57.52±8.31U),复氧180 min后钙离子荧光强度(71.13±4.74U)显著增高(P<0.01vs对照组)。而牛磺酸组细胞内钙离子荧光强度较模拟I/R组显著降低[(42.42±4.17U)vs(71.13±4.74U),P<0.01]。心肌细胞缺血/缺氧导致Ca2+超载;模拟I/R Ca2+超载加剧,而牛磺酸有明显减轻心肌细胞模拟I/R时Ca2+超载的作用。     

介质中不同Ca~(2+)浓度对五种榕树幼苗钙含量的影响

选择高榕(Ficusaltissima)、木瓜榕(F.auriculata)、聚果榕(F.racemosa)、鸡嗉子榕(F.semicorda ta)以及对叶榕(F.hispida)5种榕树,对其幼苗进行不同Ca2+(CaCl2)浓度下的水培实验,结果表明,5种幼苗钙含量随介质Ca2+浓度的增加而增加,增加趋势大致相同。高Ca2+浓度下(10mmol/L)幼苗地上部的钙含量为低Ca2+浓度下(0.05mmol/L)的1.51～1.63倍。5种榕树幼苗钙含量随培养液Ca2+浓度变化的Yadj值的多重比较结果显示,对叶榕钙含量显著高于其他四种(P<0 01),高榕则显著低于其他四种(P<0 01),木瓜榕、聚果榕、鸡嗉子榕之间的差异不显著。不同榕树幼苗地上部分钙含量受介质钙浓度影响,但可能主要决定于基因型的差异,表明榕属植物富钙基因的存在,这为高钙榕树植物资源开发提供初步的理论依据。     

Beneficial effects of taurine on mouse zygotes developing in protein-free culture medium

The objectives of this study were to determine if mouse zygotes from outbred mice can develop in simple culture medium in the absence of bovine serum albumin (BSA), and if taurine can be used as a medium supplement to improve development. Zygotes from 2 stocks of outbred mice (CD-1 and CF-1) were cultured in simple embryo culture medium (TE medium) lacking BSA and with or without taurine (24 mM), or in regular TE medium with BSA. The presence of BSA had little or no effect on development, but development to post-blastocyst endpoints was enhanced when CD-1 zygotes were cultured in medium containing taurine. In addition, when CD-1 blastocysts were transferred to pseudopregnant animals, embryos cultured in the presence of taurine developed into fetuses more often than those cultured in medium without taurine, and their weights were higher than those of embryos cultured in regular TE medium with BSA. These beneficial effects of taurine do not appear to be the nonspecific effects of a fixed nitrogen source, because the addition of glycine to BSA-free TE medium did not have similar beneficial effects. It was concluded that mouse zygotes from outbred mice do not require BSA for their preimplantation development in culture and that the presence of taurine in preimplantation culture medium is beneficial not only for preimplantation development of the zygotes, but also for their post-blastocyst development.     

Hyper-osmotic cell volume regulation in vivo and in vitro in flounder (Platichthys flesus) heart ventricles

Summary Cell volume regulation in heart ventricles of the flounderPlatichthys flesus has been studied under hyper-osmotic conditions in vivo and in vitro.During reacclimation of flounders from fresh water to sea water the osmolality of blood plasma increased to the sea water level in 7 days. The water content of the heart ventricle cells remained constant after reacclimation while the intracellular concentration of K⁺ and taurine increased to values found in sea water flounders.Hearts taken from fresh water flounders were mounted in vitro and perfused with hyper-osmotic ringer solution. Upon perfusion the cellular water content of the ventricle decreased initially but then steadily increased. After 6 h the cellular water content was still below the control values reflecting that the ventricle cells have an incomplete volume regulation in a hyper-osmotic medium. The cellular amount of K⁺ and taurine increased during the volume regulation. The perfusion media did not contain taurine or amino acids and the gain in cellular taurine must be due to intracellular production.     

Cryopreservation of brain tissue for primary culture.

Factors affecting recovery of brain cells from cryopreserved cerebral tissues of fetal rats were examined based on yields of viable cells on cell culture. Favorable preservation was obtained with freezing small pieces (less than 1 mm cube) of brain tissues rather than whole tissues or dissociated single cells, and use of 10% dimethylsulfoxide as a cryoprotectant in liquid nitrogen. As for cell preparation procedures, cell survival was improved when tissues were heated at 32 degrees C during papain digestion and centrifugation. Under favorable conditions, the number of brain cells recovered from cryopreserved tissues corresponded to 20-30% of those from fresh control tissues. Immunocytochemical characteristics of cultured neurons, astrocytes, and oligodendrocytes from cryopreserved and fresh tissues were indistinguishable. Semi-quantitive analyses of microtubule-associated protein-2 (MAP-2) and synaptophysin revealed that there was no difference in the amounts of these markers between cultures from both fresh and cryopreserved tissues. These results suggest that most of all cell types including neurons were equally susceptible to the cryopreservation procedures. We concluded that cryopreservation in liquid nitrogen is an effective method for preservation of embryonic brain tissues for later use in cell culture studies.     

The Relationship between the Effects of Cytokinin on the Expension and Metabolism of Excised Cucumber Cotyledons and Water Stress

Excised cucumber (Cucumis sativus, var. Jin-ian No. 4) cotyledons were incubated with 10 ppm of BA (benzyladenine) or water for 1 h, then thouroughly rinsed with water and grown in darkness on filter paper saturated with different concentrations of mannitol solution. Up to 24 h, the fresh weight, carotenoid, RNA, DNA and lipid contents of cotyledons were determined. Although mannitol solution reduced the effectiveness of BA treatment, in the same condition of osmotic potential, the increases of fresh weight, carotenoid, RNA and DNA contents, as well as the decrease of lipid per cotyledon were always much higher in BA treated tissues. BA enhanced the rate of water uptake by the cotyledons. The fresh weight of BA and 0.2 M mannitol treated cotyledons was equal to that of water control, but the increases of carotenoid and nucleic acid contents and the decrease of lipid were much higher in tho former than the latter. 0.3 M and 0.5 M mannitol solu- tions almost interrupted the water uptake of water and BA treated cotyledons respectively. However, the increases of carotenoid and nucleic acid contents as well as decrease of lipid were still occurred in these conditions. The different osmotic potential did nearly not affect the ratio of the increases of carotenoid and nucleic acid contents between BA treatment and control. It means the effectiveness of BA was almost the same under different osmotic potential It is evident that BA stimulated simultaneously the water uptake and metabolism of the cotyledons. They are probably different processes but closely related to each other.