109 resultados para lotus embryo


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Lancelets (amphioxus), although showing the most similar anatomical features to vertebrates, never develop a vertebrate-like head but rather several structures specific to this animal. The lancelet anatomical specificity seems to be traceable to early developmental stages, such as the vertebrate dorsal and anterior-posterior determinations. The BMP and Wnt proteins play important roles in establishing the early basis of the dorsal structures and the head in vertebrates. The early behavior of BMP and Wnt may be also related to the specific body structures of lancelets. The expression patterns of a dpp-related gene, Bbbmp2/4, and two wnt-related genes, Bbwnt7 and Bbwnt8, have been studied in comparison with those of brachyury and Hnf-3 beta class genes The temporal expression patterns of these genes are similar to those of vertebrates; Bbbmp2/4 and Bbwnt8 are first expressed in the invaginating primitive gut and the equatorial region. respectively, at the initial gastrula stage. However, spatial expression pattern of Bbbmp2/4 differs significantly from the vertebrate cognates. It is expressed in the mid-dorsal inner layer of gastrulae and widely in the anterior region, in which vertebrates block BMP signaling, The present study suggests that the lancelet embryo may have two distinct developmental domains from the gastrula stage, the domains of which coincide later with the lateral diverticular and the somitocoelomic regions. The embryonic origin of the anterior-specific structures in lancelets corresponds to the anterior domain where Bbbmp2/4 is continuously expressed.

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The possibility of the brine shrimp Artemia to produce dormant embryo (cysts) in diapause is a key feature in its life history. In the present study, we obtained a proteomic reference map for the diapause embryo of Artemia sinica using two-dimensional gel electrophoresis with a pH range of 4-7 and a molecular weight range of 10-100 kDa. Approximately 233 proteins were detected, and 60 of them were analyzed by capillary liquid chromatography tandem mass spectrometry (LC-MS/MS). Of these, 39 spots representing 33 unique proteins were identified, which are categorized into functional groups, including cell defense, cell structure, metabolism, protein synthesis, proteolysis, and other processes. This reference map will contribute toward understanding the state of the diapause embryo and lay the basis and serve as a useful tool for further profound studies in the proteomics of Artemia at different developmental stages and physiological conditions.

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郁金香属(Tulipa L.)是世界著名的观赏植物,广泛分布于欧洲、亚洲的温带地区以及非洲的西北部,中亚地区是其分布和多样化中心。该属包括老鸦瓣组、长柱组、郁金香组、毛蕊组、扭药组、鸡冠组和无毛组共七个组,40至150种。老鸦瓣组是东亚特有类群。中国共有郁金香属植物16种,主要分布于西北(新疆)以及中东部地区,其中老鸦瓣组有4个种,占该组全部种类的4/5。长期以来,由于对老鸦瓣组的生物学特性及其地理分布缺乏了解,该类群的归属问题一直都是该属系统学研究中争论的焦点之一。因此,本文以分布于我国的郁金香属作为主要研究对象,通过对其形态学、胚囊发育的比较胚胎学以及分子系统学研究,对老鸦瓣组的系统位置以及上述特征在该属分类中的意义进行了探讨。主要结果如下 1)通过对该属18种植物(包括土耳其的3个种)28个形态性状数据的分支分析,表明广义郁金香属并不是一个单系类群。Tulipa sect. Amana与该属其他四个组(sect. Orithyia、sect. Eriostemones、sect. Leiostemones和sect. Tulipanum)在分支树上各成一支,它们与Lloydia属构成一个大支的三个分支。同时,sect. Amana具有与子房近等长的花柱以及2-3(-4)个苞片等与郁金香属不同的形态特征。因此,我们认为sect. Amana应从广义郁金香属中独立出来,恢复其老鸦瓣属Amana Honda作为属的分类地位。 2)发现了一个新种:Amana kuocangshanica D.Y. Tan et D. Y. Hong。该种与A. erythronioides 和A. anhuiensis近缘,区别在于鳞茎皮内侧光滑无毛,下部叶披针形,自基部向上2/3处最宽,果喙长0.64±0.08 cm。 3)对16种植物叶表皮形态观察的结果表明,老鸦瓣属4个种的叶上表皮细胞为矩形或矩圆形、下表皮为菱形或矩形,垂周壁为直线形或波形,上表皮无气孔或气孔密度较小,这些特征与狭义郁金香属的种差异显著。在狭义郁金香属中,叶表皮特征在种间差异明显,可以作为分种及种间亲缘关系探讨的依据,但在组间没有明显的差异。 4)对19种植物的花粉形态观察表明,Amana属的4种为近椭球形、舟形和肾形, 外壁纹饰网状,网脊浅皱波状,与狭义郁金香属的15种具明显不同。在狭义郁金香属中,花粉外壁纹饰虽然在种间存在一定程度的差异,但对组的划分意义不大。 5)从种皮形态及微形态特征看,在所观察的16种植物中,Amana属的种子小,呈半月形,较厚,种柄明显,胚不易见;种皮纹饰为皱波状或不规则,与狭义郁金香属存在显著的差异。狭义郁金香属的12种在种皮纹饰、网眼大小及形状、网脊宽窄等方面均存在明显的差异,但组间无明显差别,说明这些特征在种的划分上具有一定的分类学意义。 6)对16种植物的胚囊发育过程观察表明:共有6种胚囊发育类型,即Fritillaria type、Drusa type、Tulipa iliensis type、Tulipa tetraphylla type、 Adoxa type和Eriostemones type。其中Tulipa iliensis type为所发现的一种新的胚囊发育类型。Tulipa iliensis、T. heterophylla和T. heteropetala3个种具有两种胚囊类型。在郁金香属中,胚囊的发育类型具有一定的系统学意义。 7)通过对21种郁金香以及猪牙花属2种植物基于nrDNA ITS区和cpDNA trnL-F 区的序列分析,发现广义郁金香属并非一单系类群,老鸦瓣属为猪牙花属的姐妹群。在狭义郁金香属中,sect. Orithyia、sect. Tulipanum以及sect. Eriostemones得到了该分子系统学分析的支持,而sect. Leiostemones是否成立及其系统关系问题尚有待于进一步研究。

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1.水稻多卵卵器的起源:被子植物的卵器中通常只有一个卵细胞。我们在水稻多胚品系胚囊中观察到二卵卵器和三卵卵器,本研究对其大孢子发生和胚囊发育进行了细胞胚胎学观察,揭示了水稻多卵卵器的起源.观察结果表明,该品系能进行正常的大孢子发生。大孢子母细胞进行正常的减数分裂形成四个大孢子靠近合点端的大孢子发育,其它三个退化。功能大孢子第一次有丝分裂后两个子核被一中央大液泡分隔在胚囊珠孔端和合点端,与此同时胚囊出现不均衡生长,珠孔端迅速膨大,合点端几乎不增大,致使二核末期的胚囊呈倒梨形.紧接着发生第二次有丝分裂,合点端核分裂时纺锤丝与胚囊纵轴平行,而珠孔端核分裂时纺锤丝与胚囊纵轴成4 5度夹角.由此产生的四核胚囊中,合点端一核向胚囊中部或中上部(胚囊珠孔端)迁移,四核胚囊再经一次有丝分裂形成两种类型的核分布偏离蓼型的八核胚囊。一种类型是珠孔端四个核,中部与合点各二个核,在胚囊细胞化过程中,珠孔端四核 分化成四细胞卵器,其中卵细胞和助细胞各二个,中部的二核分化成二极核中央细胞,合点 端的二核形成反足细胞。另一种类型是珠孔端六个核,合点端二个核,在胚囊细胞化过程中, 两端各一核向中部迁移分化成二极核中央细胞,珠孔端剩余的五核分化成五细胞卵器,其 中卵细胞三个,助细胞二个,合点端的一核迅速分裂形成反足细胞. 2.水稻同源三倍体TAR的生殖特性:TAR的单穗结实率平均可达10%,核型分析表明此三倍体产生的后代个体仍为具有36条染色体的三倍体.细胞胚胎学初步观察显示TAR为一具兼性无融合生殖特性的水稻新种质,其胚珠几乎都能进行胚囊的分化,但其中仅有33%的胚囊有较正常的结构,9%的胚囊在散粉前进行胚胎发生,58%的胚囊发育显著异常,表现为极性紊乱、多极核或缺失雌性生殖单位等。 3.水稻亚种间杂种败育的细胞学基础:对普通栽培稻不同品种类型间杂种颖花败育的细胞学基础及雌性败育的过程进行的细胞学研究表明:1)引起杂种颖花败育的原因有胚囊败育,花粉败育、开花时花药不开裂和雌雄异熟.其中胚囊败育而丧失受精能力是引起低结实率的最重要的因素,开花时花药不开裂和雌雄异熟在一定程度上形成了雌雄性细胞时间和空间的隔离屏障。2)杂种植株的所有大孢子母细胞都能进行正常的减数分裂形成四个大孢子,败育主要发生在靠近合点端的功能大孢子分化形成胚囊的早期,有的胚囊母细胞在进行第一次有丝分裂前便萎缩解体,多数能完成一次或二次有丝分裂形成二核或四核败育胚囊.败育的共同特征是无液泡的分化,细胞质少或退化,在败育胚囊残迹部位,解体的珠心细胞和萎缩的胚囊残溃混杂垛叠.已受精的杂种子房没有观察到胚及胚乳发育的异常.籼粳杂种胚囊败育频率较高. 4.籼粳杂种生殖障碍的基因定位:应用具有1 37个标记位点的籼粳杂交窄叶青8号/京系17)F1花药培养获得的127个双单倍体OH)群体构建的R FLP图谱,对控制籼粳杂种颖花败育的基因座位进行了定位研究。结果在第1、3、4、5、6、7、8、1 2染色体上检测到1 0个基因座位,其中第3、12染色体上的2个不育基因位点str3和str12与同一杂交组合F2分离群体中发现的异常分离热点处于相同的染色体区段.stj-6的基因加性效应为负值,有增加籼粳亲和性的作用;其余的不育基因座位皆有增加籼梗杂种不育性的作用. 5.籼粳杂种胚囊败育的遗传分析和基因定位:利用DH系构建的分子图谱及DH系衍生的2个回交群体定位了引起籼梗杂种胚囊败育的2个互补的主效基因esa-l(E1或e1位点)和esa-2(E2或e2位点),它们分别位于第6和第1 2染色体.在不育基因位点,籼稻基因型为EIEle2e2,粳稻基因型为elelE 2E 2,杂交后代中基因型为EIE2,Ele2、elE 2的雌配子体正常发育,携带ele2基因型的雌配子体表现败育.胚囊育性受配子体基因型控制,孢予体遗传背景影响胚囊败育基因的表达.

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本文中以水稻为实验材料,用共聚焦扫描显微镜检术(CLSM)和酶法分离技术观察其胚囊取得好的效果,主要实验结果如下: 1.水稻子房分别用FAA和4%戊二醛固定液固定,经乙醇脱水,然后用冬青油整体透明及丁香油封片。在共聚焦扫描显微镜下,发育中的水稻胚囊显示自发荧光,可分辨出胚囊内部结构。FAA和4%戊二醛两种固定液效果不同,后者效果更佳。用上述实验程序进一步观察拟南芥菜胚囊及水稻和拟南芥菜花粉表明,CLSM可用于快速观察分析水稻和拟南芥菜胚囊发育过程以及水稻胚囊败育情况,但不适用于观察分析其花粉发育过程。 2.水稻子房用FAA、FPA或卡诺氏固定液固定,经乙醇转至蒸镏水,然后置于由纤维素酶和果胶酶组成的酶液中,在37℃保温条件下进行连续振荡酶解,在振荡过程中辅以手工显微操作,由此即可分离出水稻固定胚珠的胚囊。用相差显微镜观察分离的胚囊表明,胚囊整体性及立体感强,观察效果好。在成功分离水稻固定材料胚囊的基础上,初步尝试了水稻新鲜材料胚囊的分离。

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植物顶端分生组织中干细胞数量的维持对于侧生器官的发生至关重要。在干细胞的基因调控网络中WUSCHEL (WUS) 是一个关键成员,围绕该基因形成两个反馈调节环,控制分生组织中干细胞群的平衡。   论文分析了用激活标签法 (activation tagging) 获得的突变体sef (stem-ecotopic-flowers),其最大的表型特点是花序轴上产生异位花和幼苗下胚轴增长。本论文就此两个表型产生的机理进行了探索,以期了解WUS基因的新功能。   对sef的表型观察发现异位分生组织不仅在花序轴上出现,而且也出现在叶柄、叶片、托叶叶腋内、花梗、花梗腋内以及花器官上。组织切片结果表明花序轴上的异位分生组织起源于已经分化的皮层细胞。对突变体的分子鉴定证明T-DNA是以单拷贝插入到WUS起始密码子上游810 bp处。对插入位点上下游各10 kb的4个基因在花序轴中的表达水平进行了分析,结果表明只有WUS基因的表达量升高,说明增强子只对WUS基因发挥了激活作用,暗示了WUS基因过表达与异位花之间存在某种联系。转35S::WUS的拟南芥幼苗下胚轴与根部出现异位的生长点;WUS被诱导表达的突变体pga6-1花序轴上出现异位花芽,证实sef的表型是由WUS超表达所导致。利用组织原位杂交和RT-PCR分析了WUS、CLAVATA3 (CLV3)、LEAFY (LFY) 与AGAMOUS (AG) 在异位分生组织中的表达模式与表达水平,结果表明WUS、CLV3、LFY、AG在花序轴表皮以下皮层中异位表达。这些结果表明WUS能激活CLV3异位表达,从而在已经分化的皮层中重新产生具有分生组织特征的细胞,同时WUS异位激活AG的表达并使LFY也在这些异位的分生组织中表达,这些分生组织发育方向被LFY与AG所决定,最终发育为异位花器官。   sef突变体另外一个突出的表型是幼苗的下胚轴增长。对幼苗期下胚轴以及胚胎4个时期的胚干细胞数进行统计,结果表明下胚轴与胚干细胞数目都呈现出sef比野生型多而wus-1比野生型少的趋势,因此sef幼苗下胚轴增长是由于细胞数目改变引起的。进一步分析发现这种区别是由于胚胎早期(授粉后1~3天)胚干细胞分裂速率的差异所造成的。利用基因芯片杂交分析突变体的基因表达谱,结果发现许多与细胞分裂相关的基因在sef中表达水平升高。RT-PCR证实这些基因在胚胎时期的表达水平升高,说明胚胎早期胚干细胞分裂速率的不同导致了幼苗下胚轴的异常。   综上所述,我们的研究结果揭示了sef异常表型的产生的可能机制。在已经分化的皮层中激活标签介导的WUS超表达激活干细胞标志基因之一CLV3和花器官基因AG,并使LFY异位表达,重新产生具有分生组织特征的细胞,这些分生组织的发育方向被LFY和AG所决定,最终发育为异位花。在sef的早期胚胎中,WUS表达增强使细胞分裂相关基因表达水平升高、细胞分裂增快,说明WUS与细胞周期相关基因的调控存在某些联系。   本论文的创新之处在于首次提出WUS表达增强能在分化的组织中产生具有分生组织特征的细胞以及WUS调控细胞分裂的结论。   

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三峡库区一些污染严重的工业企业是当地主要的点源污染源,对周围植物群落产生了巨大危害,而植物是生态系统赖以存在的基础,它的生长发育直接影响到生态系统的结构及其正常功能的实现。本研究按照与点源污染源的距离为梯度,通过在三峡库区兴山县白沙河磷化工厂周围布设了32个植物群落固定样地,并以点源污染无法影响到的植物群落为对照,进行样地的野外群落学调查;在每个样地取不同种植物叶片100克左右和样地0~20cm土壤500克,以石灰滤纸法同步进行大气氟化物的取样。样品带回室内应用氟离子选择电极法,测定大气氟化物含量、植物叶片氟的累积量和土壤水溶性氟的含量。同时在野外调查时使用PAM2100叶绿素荧光仪测定植物最大光化学效率即Fv/Fm的值。通过野外调查试验和相关的室内分析,研究了(1):点源污染对三峡库区陆生植物群落组成和物种多样性的影响;(2):点源污染中的主要污染物对植物及土壤环境的影响;(3):不同物种叶片最大光化学效率Fv/Fm对污染胁迫响应的差异。结果如下: 点源污染对植物群落物种丰富度以及Pielou均匀度指数均有不同程度的影响,对于群落结构相对简单的马尾松林和柏木林的不利影响更为显著。相对于污染区来说,对照区中物种重要值的集中程度有所下降。许多物种的重要值在污染区与对照区有明显的变化。马尾松(Pinus massoniana)、檵木(Loropetalum chinense )、铁仔(Myrsine africana)、卷柏(Selaginella tamariscina )等对照区重要值较污染区为高,黄檀(Dalbergia hupeana )、菱叶海桐(Pittosporum truncatum)、君迁子(Diospyros lotus)等污染区的重要值较对照区为高。群落中有些物种比如柏木(Cupressus funebris)的天然更新也受到污染的影响。 污染区土壤pH值大多低于对照区,但是与离污染源距离的相关性不强。污染区有些物种比如马尾松、柏木等叶片中的全氟含量与大气中氟化物的含量和土壤水溶性氟含量明显正相关。但是另外有些物种氟的累积量受点源污染的影响不显著,比如菱叶海桐、翅柃(Eurya alata)等在污染严重的样地内生存状况仍然很好。 在距离点源污染近的样地内,大多数物种的最大光化学效率Fv/Fm的值显著下降。栓皮栎(Quercus variabilis)、马尾松和柏木等的Fv/Fm值与距污染源的距离呈明显的正相关,都是随着离污染源越来越近而逐渐降低。根据污染区相对对照区Fv/Fm值下降幅度的不同,把植物划分为三种类型:对污染敏感型如柏木、铁仔、檵木等,中等敏感型如油桐(Vernicia fordii)、香叶树(Lindera communis)和不敏感型如山胡椒(Lindera glauca)和 蝴蝶花(Iris japonica)等。

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BACKGROUND: Effects of 17beta-estradiol and progesterone on rhesus monkey oocyte maturation in vitro were evaluated by embryo development subsequent to IVF. METHODS AND RESULTS: In experiment 1, immature cumulus-oocyte complexes collected from unstimulated adult females during the non-breeding season were matured in modified medium CMRL-1066 containing various combinations of gonadotrophins (FSH + LH), estradiol and/or progesterone. Formation of morulae and blastocysts was greatest in oocytes matured in medium containing estradiol and/or progesterone, with or without gonadotrophins (morula 38-46%, blastocyst 14-20%) than in control oocytes matured without estradiol or progesterone (morula 14%, blastocyst 0%). In experiment 2, cumulus-oocyte complexes from unstimulated prepubertal female monkeys were matured in medium with gonadotrophins, estradiol or progesterone. The best development to the morula stage was obtained with oocytes matured with gonadotrophins and estradiol or gonadotrophins and progesterone (43 and 25 morulae, respectively), while control oocytes matured with gonadotrophins but without steroid hormones gave the poorest morula developmental response (12%). However, there was no difference in blastocyst development across all groups (0-3%). CONCLUSIONS: These results demonstrate that during rhesus monkey oocyte maturation in vitro: (i) estradiol or progesterone can improve oocyte developmental competence; (ii) immature oocytes from prepubertal versus adult females have differential responses to challenge with estradiol or progesterone.

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The giant panda skeletal muscle cells, uterus epithelial cells and mammary gland cells from an adult individual were cultured and used as nucleus donor for the construction of interspecies embryos by transferring them into enucleated rabbit eggs. All the three kinds of somatic cells were able to reprogram in rabbit ooplasm and support early embryo development, of which mammary gland cells were proven to be the Lest, followed by uterus epithelial cells and skeletal muscle cells. The experiments showed that direct injection of mammary gland cell into enucleated rabbit ooplasm, combined with in vivo development in ligated rabbit oviduct, achieved higher blastocyst development than in vitro culture after the somatic cell was injected into the perivitelline space and fused with the enucleated egg by electrical stimulation. The chromosome analysis demonstrated that the genetic materials in reconstructed blastocyst cells were the same as that in panda somatic cells. In addition, giant panda mitochondrial DNA (mtDNA) was shown to exist in the interspecies reconstructed blastocyst. The data suggest that (i) the ability of ooplasm to dedifferentiate somatic cells is not species-specific; (ii) there is compatibility between interspecies somatic nucleus and ooplasm during early development of the reconstructed egg.

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Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo. (C) 2010 Elsevier Inc. All rights reserved.

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鸡胚胎是一种被广泛应用于发育生物学研究的实验材料。在以鸡胚胎为动物模型的各项研究工作 中, 心率常被看作是一个很重要的反应胚胎生理活动指标。本文详细介绍了一种侵入性的心电记录新方法, 在 正常的生理条件下通过记录鸡胚胎心电的变化来监测心率。首先在蛋壳上钻孔, 将电极插人蛋内, 然后通过放 大器放大, 板转换, 将心电信号输人电脑进行分析处理, 提取与心率相关的信息。这种记录方式对胚胎损 伤较小, 不影响胚胎的正常发育具有灵敏度高, 操作简单, 容易掌握等特点。

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The cryopreservation of oocytes has been only marginally successful with any of the current protocols, including slow cooling, rapid cooling and vitrification. We wished to test the hypothesis that oocytes from a single mouse strain would freeze successfully by 1 of the 3 mentioned protocols. Unfertilized Kunming mouse oocytes obtained 14 h after PMSG/hCG administration were randomly assigned to be cryopreserved after slow cooling, ultra rapid cooling and vitrification. Oocytes were thawed by straws being placed into 37 degrees C water, and their morphological appearance and in vitro fertilization capability were compared with that of oocytes that had not undergone cryopreservation. Survival of oocytes was indicated by the absence of darkened ooplasm or by broken membranes or zona pellucida. Functional integrity was evaluated by the formation of a 2-cell embryo after IVF. Survival rate of slow cooled oocytes did not differ from that seen in vitrified oocytes (55.1 vs 65.9%) but was significantly lower in the rapidly cooled oocytes (24.2%; P<0.01). The results of NF of slow cooled and vitrified oocytes were similar to those of the control group (72 and 73 vs 77%; P>0.05). It appears that Kunming mouse oocytes can be successfully cryopreserved using the slow cooling method with 1,2-propanediol and vitrification, which contains both permeating and nonpermeating cryoprotectants. (C) 1997 by Elsevier Science Inc.

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We investigated memory impairment in newly hatched chicks following in ovo exposure to a 50-Hz magnetic field (MF) of 2 mT (60 min/day) on embryonic days 12-18. Isolated and paired chicks were used to test the effect of stress during training, and memory retention was tested at 10, 30, and 120 min, following exposure to a bitter-tasting bead (100% methylanthranilate). Results showed that memory was intact at 10 min in both isolated and paired chicks with or without MF exposure. However, while isolated chicks had good memory retention levels at 30 and 120 min, those exposed to MF did not. The results suggest a potential disruption of memory formation following in ovo exposure to MF, with this effect only evident in the more stressed, isolated chicks. Bioelectromagnetics 31:150-155, 2010. (C) 2009 Wiley-Liss. Inc.

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本试验以奶水牛为研究对象,探讨卵胞质内单精子注射(ICSI)操作液中添加不同浓度聚乙烯吡咯烷酮(PVP)、细胞松弛素(CB)、透明质酸(HA)对早期胚胎发育的影响.旨在探寻一套适合奶水牛ICSI的操作程序,为高效生产奶水牛体外胚胎提供科学依据.结果表明:4%PVP组卵裂率高于6%PVP组和8%PVP组(65.8%vs59.5%、44.4%,P<0.05):4%PVP组囊胚率高于6%PVP组(21.0%vs16.2%,P<0.05)和8%PVP组(21.0%vs8.3%,P<0.01);添加CB组的卵裂率高于未加CB组(73.3%vs68.9%,P>0.05),添加CB组囊胚率高于未加CB组(22.2%vs20.0%,P<0.05);操作液中添加1.5mg/mL HA更有利于胚胎的发育.1.5mg/mL HA组和4%PVP组相比较,操作过程中润滑作用相似,HA组卵裂率略高于PVP组(71.0%vs65.8%,P<0.05);HA组囊胚数略高于PVP组(22.6%vs21.0%,P>0.05).