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1.
Bielanski A. Simard C. Maxwell P. Nadin-Davis S. 《Veterinary research communications》2001,25(8):663-673
The association of bovine immunodeficiency virus (BIV) with embryos derived by in vitro fertilization from oocytes of experimentally infected heifers or oocytes/embryos exposed to the virus in vitro was investigated. Using a nested-PCR assay, proviral DNA of BIV was not detected in follicular fluid or in embryos derived from BIV-infected donors. In vitro exposure of oocytes to BIV during maturation or insemination with BIV-infected semen resulted in zona pellucida-intact embryos testing negative for BIV provirus. However, exposure of zona pellucida-free day-7 embryos to the virus resulted in a positive BIV assay for 28% of the batches of embryos, suggesting that the zona pellucida has a role in protecting against BIV infection. The presence of BIV in the IVF system had no apparent effect on the development of bovine embryos to the blastocyst stage. 相似文献
2.
一个重要的精子受体—猪透明带糖蛋白β-D-半乳糖残基 总被引：1，自引：0，他引：1
【目的】证明末端半乳糖残基在猪透明带糖蛋白中的分布以及它在精子结合和侵入中的作用。【方法】体外成熟的猪卵母细胞去除周围卵丘细胞，用异硫氰酸荧光素(fluorescein isothiocyanate，FITC) 络合西非单叶豆凝集素(Bandeiraea simplicifolia lectin-I，BS-I) 或蓖麻凝集素(Ricinus communis agglutinin，RCA-I)处理30min，分别观察末端-D-半乳糖残基或-D-半乳糖残基在透明带中的定位分布。荧光显微镜观察显示，BS-I和RCA-I都标记于整个透明带上且比较集中于透明带的外层，其中BS-I显示微弱荧光反应，而RCA-I呈强荧光反应且在透明带的外层反应尤为强烈。植物凝集素处理的卵母细胞用于体外受精，分别在受精2h或12h观察精-卵结合和精子入卵情况。【结果】RCA-I处理组卵母细胞平均精子结合数显著低于对照组（18 vs 95），而精子入卵则完全被阻断；可是BS-I处理组的卵母细胞的精子结合数少量被抑制（59 vs 95），而且精子入卵率与对照组无显著差异。【结论】猪透明带末端-D-半乳糖残基作为精子受体的重要组成成分，参与精-卵结合和受精。 相似文献
3.
Fuminori TANIHARA Michiko NAKAI Hiroyuki KANEKO Junko NOGUCHI Takeshige OTOI Kazuhiro KIKUCHI 《The Journal of reproduction and development》2013,59(4):385-392
In porcine oocytes, the function of the zona pellucida (ZP) with regard to sperm
penetration or prevention of polyspermy is not well understood. In the present study, we
investigated the effects of the ZP on sperm penetration during in vitro
fertilization (IVF). We collected in vitro-matured oocytes with a first
polar body (ZP+ oocytes). Some of them were freed from the ZP (ZP− oocytes) by two
treatments (pronase and mechanical pipetting), and the effects of these treatments on
sperm penetration parameters (sperm penetration rate and numbers of penetrated sperm per
oocyte) were evaluated. There was no evident difference in the parameters between the two
groups. Secondly, we compared the sperm penetration parameters of ZP+ and ZP− oocytes
using frozen-thawed epididymal spermatozoa from four boars. Sperm penetration into ZP+
oocytes was found to be accelerated relative to ZP− oocytes. Thirdly, we evaluated the
sperm penetration of ZP+ and ZP− oocytes at 1−10 h after IVF (3 h gamete co-incubation).
The proportions of oocytes penetrated by sperm increased significantly with time in both
groups; however, the number of penetrated sperm per oocyte did not increase in ZP−
oocytes. Finally, we performed IVF using ZP− oocytes divided into control (3 h) and
prolonged gamete co-incubation (5 h) groups. Greater numbers of sperm penetrated in the 5
h group than in the control group. These results suggest that the ZP and oolemma are not
competent factors for prevention of polyspermy in our present porcine IVF system. However,
it appears that ZP removal is one of the possibilities for reducing polyspermic
penetration in vitro in pigs. 相似文献
4.
为探究新娘草叶片对光照变化的形态建成和超微结构的响应机制,以新娘草茎段扦插苗为试验材料,通过遮阳网人工模拟5种光环境(遮光度分别为0、25%、50%、75%、95%)进行试验,研究不同光照强度对新娘草叶片生长及超微结构的影响。结果表明:随光强的减弱,新娘草叶长、叶宽和叶面积呈先增后减的变化趋势。在遮光75%时,叶长、叶宽和叶面积达到峰值,叶面颜色由黄绿色变为浓绿色,叶背颜色由深紫色变为浅紫色,在遮光95%时,紫色消失。光强对叶绿素a(Chl a)、叶绿素b(Chl b)、叶绿素a+b[Chl (a+b)]和花色素苷(Ant)含量均有显著影响(P<0.05):Chl a、Chl b和Chl (a+b)含量与光强呈负相关,Ant含量与光强呈正相关。新娘草叶片厚度、上表皮厚度、下表皮厚度、栅栏组织厚度、海绵组织厚度均随光强的降低而显著降低(P<0.05),而栅栏/海绵组织厚度和组织密实度均在50%遮光下达到峰值。全光及遮光25%条件对叶肉细胞的超微结构产生了轻微的破坏,造成部分基粒类囊体片层和基质类囊体片层的扩张,淀粉粒体积变大,而极端弱光(遮光95%)对叶肉细胞的超微结构产生了巨大的破坏,使得部分叶绿体解体,淀粉粒空洞。研究结果表明新娘草具有较强的光适应性,可以通过调整叶形、叶色、叶片解剖结构的方式适应不同的光环境,适度的光照强度(遮光50%~75%)有利于新娘草维持最佳的生长状态,而在极端弱光环境(遮光95%)下新娘草虽能存活,但叶片内部器官的结构产生了不可逆的损伤。 相似文献
5.
为比较猪卵母细胞在GV期与MⅡ期的冷冻保存效果,试验在这两个成熟阶段对其进行玻璃化冷冻,GV期卵母细胞解冻后培养至成熟,MⅡ期卵母细胞解冻后恢复2 h,然后采用免疫荧光标记、Western blotting和链霉蛋白酶溶解方法分别检测它们的皮质颗粒分布、CD9蛋白表达水平和透明带消化时间上的差异。结果表明,GV期卵母细胞在解冻后2 h的存活率显著低于MⅡ期卵母细胞（P<0.05）,但极体排出率与对照卵母细胞无明显差异（P>0.05）;在冷冻MⅡ期卵母细胞中,皮质颗粒的皮质区分布比例和CD9的蛋白表达水平显著下降（P<0.05）,但冷冻GV期卵母细胞经体外成熟后则无明显变化（P>0.05）;冷冻GV期与MⅡ期卵母细胞均不会影响透明带的消化时间（P>0.05）。由此可见,猪卵母细胞在GV期的冷冻存活率虽然较MⅡ期低,但其体外成熟后极体排出率、皮质颗粒分布和CD9蛋白表达水平均未受到冷冻的影响。 相似文献
6.
Tomohiro SASANAMI Mamoru OHTSUKI Yukio DOI Ryota TANAKA Atsushi FUJISHIMA Makoto MORI 《Animal Science Journal》2008,79(1):104-110
The extracellular matrix surrounding the oocyte before ovulation is called the perivitelline membrane (PL) in avian species. The PL is constructed with two major glycoproteins, ZPC and ZP1, which are synthesized in the ovarian granulosa cells and the liver, respectively. Although the properties of the major components in the PL have been examined, knowledge about the nature of its minor constituents is lacking. In this study we focused on PL protein, which migrates at 46‐kDa in the gel of SDS‐PAGE. N‐terminal sequence analysis demonstrated that the 46‐kDa protein is the C‐terminal fragment of ZP1. Analysis of lysylendopeptidase digests or cyanogens bromide‐degraded fragments of ZP1 confirmed this postulate. Western blot analysis using antiserum against 46‐kDa protein indicated the absence of 46‐kDa protein in the serum. Moreover, small immunoreactive bands, thought to be cleaved fragments of ZP1, were detected in the PL lysate by western blot analysis using antiserum against the N‐terminal peptide of ZP1. These results indicated that the N‐terminal proteolytic processing of ZP1 might take place after the arrival of ZP1 at the ovary, and the resulting product, 46‐kDa protein, is incorporated into the PL. 相似文献
7.
本研究旨在探究猪卵母细胞体外成熟过程中添加泛素结合酶(E2)抑制剂对卵母细胞透明带蛋白泛素化水平及精-卵结合能力的影响。试验分为6组:对照组、DMSO组、5、10、15和20μmol·L-1 NSC697923处理组。采用Western blot方法分析体外成熟培养液中添加不同浓度泛素结合酶抑制剂NSC697923对猪卵母细胞透明带泛素化水平表达的影响。通过Hoechst染色检测不同组别的精卵结合能力。结果表明:1)猪卵母细胞体外成熟培养液中,添加10、15和20μmol·L-1 NSC697923显著降低卵母细胞成熟率(P<0.05),透明带硬化时间(P<0.05)以及精卵结合率(P<0.05)。2)对照组和各处理组的透明带蛋白在61、81、106 ku处发生不同程度的泛素化标记,而添加15和20μmol·L-1 NSC697923显著地降低透明带蛋白泛素化水平(P<0.05)。综上表明,猪卵母细胞体外成熟过程中泛素结合酶(E2)改变成熟卵母细胞透明带泛素化水平以及精卵结合能力。 相似文献
8.
A.M.H. Abdel Dayem Karima Gh. M. Mahmoud M.F. Nawito M.M. Ayoub T.H. Scholkamy 《Livestock Science》2009,122(2-3):193-198
This study aimed to develop a system of in vitro assays based on zona pellucida binding and in vitro fertilization for predicting male fertility in buffalo bulls. Frozen–thawed semen from nine bulls was tested for motility, viability index, acrosomal integrity, zona pellucida binding and in vitro fertilizing ability. Differences in post-thaw sperm motility between bulls were not significant. Differences in viability indices and percentage of spermatozoa with detached acrosome between bulls was highly significant (P < 0.001). Sperm attached per ovum, fertilization rates and polyspermy percentages varied significantly (P < 0.01) among buffalo bulls. A significant (P < 0.01) positive correlation coefficient of 0.69 was evident between normal acrosome and sperm attached per ovum, while between normal acrosome and fertilization efficiency it was 0.72. Sperm from different buffalo bulls differs in their ability to bind and fertilize oocytes. This study provides a basis to predict and maximize the in vitro fertilization performance of individual bulls. 相似文献
9.
10.
This study was designed to compare the efficiency of porcine oocytes vitrified at the GV and MⅡ stages. The vitrified GV oocytes were matured in vitro and then evaluated their cortical granule distribution with immunofluorescence, CD9 protein level with Western blotting and zona pellucida dissolution time with pronse digestion method, as compared to vitrified MⅡ oocytes.The results showed that the survival percentages of oocytes vitrified at the GV stage were significantly lower than those vitrified at the MⅡ stage after 2 h of warming (P<0.05);However, there was no difference in maturation rate between vitrified and fresh oocytes (P>0.05). The oocytes vitrified at MⅡ stage resulted in significantly decreased normal cortical granule distribution and CD9 protein level (P<0.05), but no such results were found in vitrified GV oocytes after maturation (P>0.05). In addition, vitrification did not affect the dissolution time of zona pellucida in GV and MⅡ oocytes (P>0.05). The data demonstrated that despite of lower survival rate compared with MⅡ oocytes, vitrified porcine GV oocytes after maturation showed normal polar body extrusion, cortical granule distribution and CD9 protein level. 相似文献