猪卵巢卵母细胞的体外成熟和体外受精   总被引：8，自引：0，他引：8  

邓满齐  谢成侠 《畜牧兽医学报》1992,23(1):1-7

将从屠宰母猪卵巢采得的卵母细胞-卵丘细胞复合体(Oocyte-cumulus Cell Complex.OCC)在含PMSG的M199培养40～44小时,卵丘细胞大部分扩散(86.4％)。48.1％(142/295)的卵母细胞排出第一极体(PBI)。将体外成熟的卵母细胞与体外获能精子授精后30～70小时,80.5％(103/128)的卵母细胞受精并可在体外发育到2～8细胞甚至桑椹胚。本文还对裸卵母细胞的体外成熟和体外受精进行了研究,对体外受精卵的早期发育作了观察。实验结果表明:PMSG对诱导卵丘细胞扩散及卵母细胞的全面成熟有重要作用,在OCC中的卵母细胞成熟率高于裸卵母细胞体外授精后8～10小时将受精卵放入改良KRB液培养可使卵裂比例明显提高。  相似文献   

A pre-synchronization program at early postpartum might increase the chances of Bos indicus cows cycling prior to 50 days regardless of the length of calf separation     

Libia PéREZ-TORRES  Ivette RUBIO  Manuel CORRO  Abraham COHEN  Agustín ORIHUELA  Carlos S. GALINA  J. Luis PABLOS 《The Journal of reproduction and development》2015,61(3):199-203

The aim of this study was to establish if pre-synchronization would enhance the number of animals cycling prior to conventional breeding at 45 days irrespective of the length of calf separation. Multiparous Bos indicus cows were allotted in four groups (n = 10). Control group (C) dams remained with their calves; groups G24, G48 and G72, which were partially weaned for 24, 48 and 72 h, respectively, were estrus synchronized using a controlled internal drug. These procedures were performed at 25 days and again at 45 days postpartum. The number of follicles, presence of a corpus luteum and back fat thickness (BFT) were determined by ultrasound. The proportion of cows with estrus and ovulation at day 25 postpartum was statistically different between the control and treated groups, with the values being 20, 60, 50 and 70 for the control, G24, G48 and G72 groups respectively (P < 0.05). At days 45 postpartum, the proportion of cows with estrus and ovulation was different in group G48 compared with the other groups (P <0.05). The average BFT and body condition score for the four experimental groups in the two periods were similar (P >0.05). Animals with a higher proportion of follicles from 17 to 21 mm, BFT values above 3.5 mm and a regular body condition were significantly different regardless of whether the dams remained with their calves or were separated, regardless of the length of this event. It can be concluded that (1) a pre-synchronization program at day 25 could trigger the onset of ovarian activity and facilitate a breeding program at day 50 and (2) temporary weaning enhances the effect of a pre-synchronization program.  相似文献   

Downregulation of the expression of inhibin α subunit and betaglycan in porcine cystic follicles     

Chunqiang WANG  Chunjing LI  Hongjiao LI  Wei MA  Shuxiong CHEN  Yun ZHAO  Jiahui RAO  Xu ZHOU 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(11):1419-1425

Inhibins, as members of the transforming growth factor beta (TGF-β) superfamily, downregulate the synthesis and secretion of follicle-stimulating hormone (FSH) in an endocrine manner. The role of inhibin/betaglycan in the ovary regulation recently gained attention. To date, no data exist on the function of inhibin α subunit and betaglycan in cystic follicles. In this study, the expressions of inhibin α subunit and betaglycan in cystic follicles were investigated using immunohistochemistry, real-time PCR and Western blot analysis. Both inhibin α subunit and betaglycan immunoreactivities were mainly localized in the granulosa cells of follicles. Expression of inhibin α subunit and betaglycan was inferior in cystic follicles compared with that in normal large follicles. However, the result of enzyme-linked immunosorbent assay showed no significant difference in the decreasing in concentration of inhibin α subunit in cystic follicular fluid compared with the control (P>0.05). In this study, we explored the effects of FSH on betaglycan expression in granulosa cells in vitro. As expected, a significant increase in the expressions of betaglycan mRNA and protein in granulosa cells was observed in response to exogenous FSH (30 ng/ml) (P<0.05) compared with the control. Consequently, this study provides evidence that the expressions of inhibin α subunit and betaglycan are inferior in cystic follicles, and this may be caused by the decrease in FSH in the presence of a cystic follicle.  相似文献   

Classification of differentiating oocytes during ovarian cycle in the giant freshwater prawn, Macrobrachium rosenbergii de man     

Prasert Meeratana  Prasert Sobhon 《Aquaculture (Amsterdam, Netherlands)》2007,270(1-4):249-258

Based on the light microscopic observations of cells' sizes, chromatin patterns, amount of lipid droplets and yolk granules, the female germ cells could be classified into four different phases, which include 1) oogonia (Oog), 2) primary oocytes (pOc), 3) secondary oocytes (sOc), and 4) mature oocyte (mOc). Oog are small oval-shaped cells with irregular-shaped nuclei sizing 4–6 μm in diameter. They rest on the connective tissue germinal cord at the tip of each ovarian pouch (lobule). Oogonia increase their number through mitotic division, and the daughter cells move into ovarian pouch where they undergo first meiotic division to become primary oocytes, which have various steps of 1st meiotic prophase accumulating at the innermost zone of the ovarian pouch. The primary oocytes are small oval-shaped cells (8.5–10 μm in diameter) with large nuclei containing chromatin in various states of condensation that finally transform into chromatids. Their nuclei are surrounded by thin rim of faint blue-stained cytoplasm. The secondary oocytes derived from 2nd meiosis and comprise five steps: Oc1 and Oc2, classified as previtellogenic oocytes, Oc3 and Oc4, classified as vitellogenic oocytes, and mature oocyte (mOc) The zones of ovarian pouch are defined based on the accumulation of various steps of developing oocytes, namely, oogenic, previtellogenic, vitellogenic and mature zones, respectively. The ovarian cycle is divided into five stages based on the number and types of oocytes present in each stage. Stage 0 and I are spawn and spent stages. Stage II and III are proliferative and premature stages, while stage IV is mature stage. During ovarian stage I, each ovarian pouch contains primarily oogonia, primary oocytes, Oc1 and a few Oc2. In stage II, the pouch contains mainly Oc2 and Oc3, while in stage III the predominant cells are Oc4. Mature oocytes appear synchronously, in stage IV. The ovulating mature oocytes pass through the thin disrupted wall of ovarian pouch into subcapsular space, that leads into the oviduct situated on the ventro-lateral side of the ovarian lobe. At spawning, the ovarian pouches break down and only connective sheaths and hemolymph sinuses remain. The germinal cords and islets of oogonia remain in the central area of stage 0 ovary. The ovarian capsule, including the muscular layer, becomes attenuated as the ovary progresses from stage 0 to IV. The hemolymph vessels become highly convoluted in the central area of the ovary, and they branch radially into smaller hemolymph sinuses around each oogenic pouch.  相似文献   

Modulation of sodium–bicarbonate co-transporter (SLC4A4/NBCe1) protein and mRNA expression in rat’s uteri by sex-steroids and at different phases of the oestrous cycle     

Khadijeh Gholami  Sekaran Muniandy  Naguib Salleh 《Research in veterinary science》2014

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Serotonin induces ovarian maturation in giant freshwater prawn broodstock, Macrobrachium rosenbergii de Man   总被引：1，自引：0，他引：1  

Prasert Meeratana  Boonsirm Withyachumnarnkul  Praneet Damrongphol  Kanokphan Wongprasert  Anchalee Suseangtham  Prasert Sobhon 《Aquaculture (Amsterdam, Netherlands)》2006,260(1-4):315-325

This study investigated the effects of serotonin (5-hydroxytryptamine or 5HT) on ovarian development in Macrobrachium rosenbergii de Man. Adult female prawns at the ovarian stage I (spent) were injected with 5HT at 1, 5, 10, 20 and 50 μg g^− 1 body weight (BW) intramuscularly on days 0, 5 and 10, and sacrificed on day 15. The doses as related to the effect could be categorized into three levels: low (1 and 5 μg g^− 1 BW of 5HT), medium (10 and 20 μg g^− 1 BW of 5HT) and high (50 μg g^− 1 BW of 5HT). The low-dose, especially at 1 μg g^− 1 BW, caused prawns to exhibit a significant increase in ovarian index (ovarian weight/body weight × 100) (5.79 ± 0.09%) as compared to the control (1.49%). The ovaries of most of these prawns could develop to stage IV (mature) and contained synchronously mature oocytes while most of the control ovaries remained at stage I and II (proliferative), and contained only oogonia to previtellogenic (Oc1, Oc2) and early vitellogenic oocytes (Oc3). The medium- and high-dose treated prawns exhibited ovaries that could reach stages III and IV and contained various types of oocytes of different maturity. Pretreatment with 5HT receptor antagonist, cyproheptadine (CYP), at 10 μg g^− 1 BW before 5HT injection significantly suppressed the effect of 5HT. Intramuscular injection of the 5HT-primed thoracic ganglion culture medium into CYP-pretreated prawns resulted in the increase of ovarian index about 5–6 times more than in the control, and in the groups injected with 5HT-primed media from muscle strip, eyestalk and brain. The ovaries of most prawn could develop up to stage IV and contained synchronously developed vitellogenic (Oc4) and mature oocytes (Oc5). These findings suggest that 5HT indirectly induces ovarian development and oocytes maturation in M. rosenbergii, probably via a putative ovarian stimulating factor released from the thoracic ganglia.  相似文献   

Cystic Rete Ovarii and Uterine Tube Adenoma in a Rabbit     

James K. CHAMBERS  Kazuyuki UCHIDA  Kenichiro ISE  Hiroyuki NAKAYAMA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2014,76(6):909-912

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Use of endoscopy for gender and ovarian stage determinations in Russian sturgeon (Acipenser gueldenstaedtii) grown in aquaculture     

Avshalom Hurvitz  Karen Jackson  Gad Degani  Berta Levavi-Sivan   《Aquaculture (Amsterdam, Netherlands)》2007,270(1-4):158-166

The Russian sturgeon (Acipenser gueldenstaedtii) is native to the Caspian Sea, the Black Sea and the Azov Sea. It used to breed in the main incoming rivers, until dam construction in the mid 20th century blocked upriver spawning migration. Aquaculture of Russian sturgeon has only recently begun, prompted by their declining populations in natural habitats and the rise in meat and caviar prices. However, information on their gonadal development and puberty under culture conditions is incomplete.

Because sturgeons have no external sexual dimorphism and there are no external markers for sexing, internal examination of the gonads must be employed for gender identification as well as to monitor their development. The present study describes endoscopic and histological observations of the gonads of young Russian sturgeons aimed at identifying gender and monitoring ovarian developmental stage in females up to the age of 6 years, when they enter their first puberty cycle, as well as at 7 years of age, when they have completed vitellogenesis, under culture conditions. This information, as related to fish age and size, is of vital importance to commercial farming of Russian sturgeon for caviar production and reproduction.

For gonadal observations in both sexes, we used an endoscopic system consisting of a 4 mm, 14 cm long cystoscope sheath incorporated with fiber-optic light transmission, connected to a halogen cold light source and a miniature videocamera with a control unit attached to a color monitor. This system allowed us viewing of the fish's abdominal organs, and to save pictures of selected areas of the gonads on a computer as the fish's personal record. Ovarian biopsies were taken in parallel for histology at typical stages of gonadal development.

Gender could be identified with this system as early as at 3 years of age and the sex ratio under culture conditions of females, males and unidentified gender were 55, 40 and 5%, respectively.

Not only did large differences occur in the developmental stages of female of the same age group, but also ovarian development was highly asynchronous at the early vitellogenic stages. In late vitellogenesis, at the “gray egg” stage (1600–2600 μm diameter), the oocytes were quite regular in size and color, and remained so until the final stages of maturity.

Our study suggests that endoscopy is an efficient method for both gender identification at an early age, and for determination of gonadal development stage in sturgeon aquaculture. The ability to see the whole intact gonads of anesthetized fish can reveal important management and research information, with minimal damage or stress to the fish.  相似文献   

Effect of 3 isoforms of Ikaros on proliferation of human ovarian cancer SKOV3 cells     

HE Li-cai  CHEN Shang  ZHU Zhen-feng  GAN Jun  YU Hui-min 《园艺学报》2015,31(8):1407-1411

AIM: To investigate the effect of Ikaros isoforms on the proliferation of human ovarian cancer SKOV3 cells. METHODS: Three isoforms of Ikaros, IK1, IK2 and IK6, were transfected into ovarian cancer SKOV3 cells. CCK-8 assay and cell counting were used to detect the effects of Ikaros isoforms on the proliferation of SKOV3 cells. The cell cycle was analyzed by flow cytometry. The cell cycle-related proteins were detected by Western blot. RESULTS: IK1 and IK2 expression inhibited SKOV3 cells proliferation. Flow cytometry analysis indicated that IK1 and IK2 induced SKOV3 cell cycle arrest at the G₁ phase. IK6 isoform exerted no obvious effect on the proliferation or cell cycle of SKOV3 cells. Compared with control EV group, IK1 group and IK2 group showed a dramatic elevation in the expression of the cell cycle inhibitor p21, along with a substantial decrease in the expression of the cell cycle inducers cyclin D1 and cyclin D2, which did not change in IK6 group. CONCLUSION: IK1 and IK2 significantly inhibit the proliferation of ovarian cancer SKOV3 cells and induce cell cycle arrest at G₁ phase by regulation of cell cycle-related proteins cyclin D1, cyclin D2 and p21, while IK6 isoform exerts no obvious effect on the proliferation and cell cycle of SKOV3 cells.  相似文献   

Effect of hPTTG1 siRNA on proliferation and apoptosis of ovarian cancer cells     

LIU Jie  SHEN Wen-jing  LU Yao  ZHAO Xun  SONG Min 《园艺学报》2011,27(1):102-107

AIM: To observe the proliferation and apoptosis of ovarian cancer cells by silencing the expression of human pituitary tumor-transforming gene 1 ( hPTTG1 ) using RNA interference technique.METHODS: The chemically synthesized siRNA targeting hPTTG1 was transfected into ovarian cancer cell line A2780 in vitro. The expression levels of hPTTG1 and c-myc were examined by RT-PCR and Western blotting. Cell proliferation was measured by MTT colorimetric assay and -TdR incorporation test. Cell apoptosis was detected by flow cytometry with annexin V/PI and TUNEL labeling.RESULTS: The expression of hPTTG1 at mRNA and protein levels was inhibited after transfection of hPTTG1 siRNA. The inhibitory efficiency was 70.5%±3.9% and 63.8%±4.5%, respectively. The absorbance began to decrease 24 h after transfection of hPTTG1 siRNA,and the highest inhibitory rate was 42.9%±5.2% at 48 h post-transfection. Radioactive incorporation of -TdR in hPTTG1 siRNA group was lower than that in normal and negative groups. The survival rate declined while the apoptotic rate and necrotic rate increased in hPTTG1 siRNA group. Apoptotic index in hPTTG1 siRNA group was higher than that in normal and negative groups. The expression of c-myc at mRNA and protein levels was down-regulated.CONCLUSION: Cell proliferation is inhibited and cell apoptosis is induced by hPTTG1 siRNA through down-regulating the expression of c-myc. hPTTG1 can be regarded as a candidate gene for ovarian cancer gene therapy.  相似文献