Role of IP3R in LH-EGFR-induced mouse oocyte meiotic resumption

AIM: To explore the effect of inositol 1, 4, 5-trisposphate receptor (IP₃R) in luteinizing hormone-epidermal growth factor receptor (LH-EGFR)-induced oocyte meiotic resumption. METHODS: Models of mouse cumulus-oocyte complexs (COCs) culture and follicle culture in vitro were generated to study the effects of 2-aminoethyl diphenyl borate (2-APB) and heparin (IP₃R specific inhibitors) on LH/EGF-induced oocyte meiotic resumption and EGF-induced cumulus cell expansion. Real-time PCR was used to detect the mRNA expression of cumulus expansion-related factors. The changes of the intracellular calcium level were monitored using Fluo 3-AM, and the cGMP level was measured by ELISA. RESULTS: The inhibitors of IP₃R, 2-APB and heparin, dramatically reversed EGF-induced oocyte maturation (P<0.05) and decreased cGMP levels in COCs (P<0.05). In addition, 2-APB and heparin reversed EGF-induced cumulus expansion, and significantly inhibited EGF-induced cumulus expansion-related factor expression (P<0.05). The activation of IP₃R increased intracellular calcium level, and the study found that 2-APB and heparin dramatically reversed EGF-induced elevation of calcium level in cumulus cells (P<0.05). Follicular culture in vitro showed that 2-APB and heparin significantly reversed the LH-induced oocyte maturation (P<0.05). CONCLUSION: LH-EGFR signaling pathway increases calcium level in cumulus cells through IP₃R, resulting in meiotic resumption.     

Advances in Holding and Cryopreservation of Equine Oocytes and Embryos

Methods for holding of oocytes and embryos during shipment as well as for their cryopreservation can greatly aid equine reproductive management. Oocytes can be held at room temperature overnight or at cooler temperatures for two nights without affecting maturation or embryo development after intracytoplasmic sperm injection. In contrast, methods for cryopreservation of equine oocytes that support high rates of embryo development have not yet been established. Equine embryos may be held overnight at temperatures from 5°C to 19°C without reduction in viability, but longer holding periods, or higher holding temperatures, may be detrimental. Small equine embryos (<300 μm), either in vivo derived or in vitro produced, can be slow frozen or vitrified successfully. In the last decade, methods have been developed to allow in vivo–derived expanded blastocysts, up to Day 8, to be vitrified successfully after blastocoele collapse. These methods of shipment and preservation allow mare owners in remote locations to have access to sophisticated assisted reproductive technologies.     

山羊卵泡卵母细胞体外成熟的研究

为了建立针对山羊卵母细胞的理想体外成熟培养体系,对山羊卵泡卵母细胞的采集方法,FSH和LH对山羊卵泡卵母细胞体外成熟的作用,及培养时间对卵母细胞体外成熟的影响进行了研究。结果表明:采用切割法平均每枚卵巢采集到的A,B级卵母细胞的数量明显多于抽吸法;采用TCM199添加体积分数为10?S,1~2 mg/L 17β_E2,10 mmol/L Hepes,0.12 mg/mL丙酮酸钠,0.1 mg/mL链霉素和0.125 mg/mL青霉素,10 mg/L FSH及20~50 mg/L LH的培养体系,有利于山羊卵母细胞的体外成熟,并放出第一极体。山羊卵母细胞体外成熟培养20,22,24 h,成熟率显著高于培养18,26,30 h(P<0.05)。     

哺乳动物卵母细胞孤雌生殖激活的研究进展

介绍了哺乳动物孤雌激活的方式,阐述了孤雌激活的方式机制及提高孤雌激活的技术措施。     

猪卵母细胞孤雌激活的研究

从卵母细胞孤雌激活的机制及影响因素,猪卵母细胞的激活,猪孤雌胚死亡主要原因,提高卵母细胞孤雌激活效果的技术途径,以及对孤雌激活的研究意义和发展前景等方面,对猪卵母细胞孤雌激活研究进行了综述.     

全氟辛酸对小鼠卵母细胞的毒性影响

【目的】研究全氟辛酸(Perfuorooctanoic aid,PFOA)对小鼠卵母细胞的毒性影响。【方法】选用6周龄体质量相近的昆明雌性小鼠160只,分为4组,每组5个重复,每个重复8只小鼠。在适应环境7 d后开始对小鼠灌服不同剂量的PFOA,其中,对照组、低剂量组、中剂量组和高剂量组的每日剂量分别为0、 5、 10和20 mg/kg,连续灌服14 d,然后利用孕马血清促性腺激素(Pregnant mare serum gonadotropin,PMSG)和人绒毛促性腺激素(Human chorionic gonadotropin,hCG)对各组小鼠进行超数排卵,获取卵母细胞进行检测。【结果】与对照组相比,PFOA中、高剂量组小鼠的卵母细胞成熟率分别下降了14.28%、28.17%;卵母细胞内活性氧含量分别升高了135%、177%;而且卵母细胞β-tubulin形态异常,染色体非整齐排列的比例分别升高了65.06%、75.60%。此外,低、中、高剂量组PFOA处理后小鼠的卵母细胞内磷酸化组蛋白H2AX(Phospho-histone H2A.X,P-H2A.X)比例分别比对照组升高了...     

电脉冲及6-DMAP对小鼠卵母细胞孤雌激活的研究

实验研究了不同电脉冲条件、6-DMAP作用不同时间以及二者联合使用时对注射hCG后18～19h采集的小鼠卵母细胞孤雌激活及发育的效果。结果表明:小鼠卵母细胞电激活后,放入2mmol/L6-DMAP的CZB中作用6h,其激活率和囊胚率都显著高于单独使用一种激活方法。其中场强2.0kv/cm,脉宽80μs,3次脉冲结合6-DMAP组的激活率和囊胚发育率最高,与其他两组差异显著。     

Serotonin induces ovarian maturation in giant freshwater prawn broodstock, Macrobrachium rosenbergii de Man

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.     

Pesticide—induced reproductive dysfunction in Indian fishes

Numerous studies are available reporting the effects of pesticides on reproductive activity in Indian fishes. The majority of these reports deals with histopathological changes in gonads and endocrine glands involved in the regulation of reproduction following treatment with different pesticides. Pesticides are reported to cause degenerative changes in gonads and arrest gametogenic processes either by acting directly on the gonads or by interfering with the secretory activity of the hypothalamo-hypophyseal-gonadal/thyroid axis that regulates various reproductive events. Secretion of hormones such as gonadotropin-releasing hormone (GnRH), gonadotropin, growth hormone, adrenocorticotropic hormone (ACTH), testosterone, estrogens, 17,20β-dihydroxyprogesterone and thyroid hormones are in general lowered, leading to cessation of gametogenesis, vitellogenesis, oocyte maturation, ovulation, spermiation, etc. Adverse effects of pesticides have also been demonstrated on fecundity, fertilization, hatching, and postembryonic development. The effects are highly variable and depend on the nature, dose, and mode of application of the pesticides.