Follicullar Development and Embryo Recovery Following 3 versus 8 FSH Injections in Heifers

Ovarian follicular dynamics and embryo yield were studied during 2 different FSH regimens for superovulation of cattle. Twenty heifers were given intramuscular injections of FSH (total of 35 mg NIH) either once daily for 3 days (Group 3×1) or twice daily for 4 days (Group 4×2). At 72 h after the first FSH injection, each animal was injected with 0.75 mg cloprostenol. Inseminations were performed at 12 h and 24 h after the onset of heat. Transrectal ultrasonography was performed on the day of the first FSH injection, the day of cloprostenol injection, the day of insemination and finally on the day of embryo recovery (day 6 or 7 after heat). The numbers of small (2–4 mm), medium (5–9 mm) and large (>10 mm) size follicles were recorded. The total number of corpora lutea, eggs and transferable embryos were recorded on the day of embryo recovery. No differences were found between the 2 groups in either of the parameters studied (p>0.05). It can be concluded that treatment with this FSH preparation once daily for 3 days gives a folliculogenic and superovulatory response similar to a treatment regimen where it is given twice daily for 4 days.     

PGF2αeCG及hCG 3种生殖激素联用对母犬发情、卵泡发育及胚胎着床的影响

给休情期性成熟母犬肌注 PGF2α(0 .5 m g/ kg) ,2 4 h后肌注 e CG(5 0 IU/ kg) ,再经 12 0 h后肌注 h CG(2 0 U/ kg) ,诱导其发情。将诱导发情的母犬与公犬交配 ,于交配后 16 d,摘取卵巢、子宫 ,观察其卵泡发育和胚胎着床情况。结果表明 ,药物处理可明显促使母犬提前结束休情期而进入发情期 (9/ 10 ) ,促进卵巢内卵泡发育 (P<0 .0 1) ,并使更多的胚胎着床 ,每只母犬子宫胚胎着床点为 (17.4± 2 .2 )个 ,提示 3种生殖激素联合应用可提高母犬繁殖率     

鸡促性腺激素释放激素对卵泡膜分泌雌二醇的影响

本实验选用性成熟的京白种蛋鸡,从同一产蛋顺序中取其发育不同阶段的各级卵泡(F_1～F_4),分离膜层,消化为单个的膜细胞,进行短期细胞培养,着重观察鸡促性腺激素释放激素(GnRH)对培养过程中膜细胞雌二醇分泌的影响.用放射免疫法测定细胞培养液中雌二醇的含量,得到以下结果:①未加外源激素处理的对照组细胞,随着卵泡从小到大的发育成熟过程,膜细胞雌二醇的分泌量逐渐降低;②适当剂量的鸡 GnRH-Ⅱ对各级卵泡膜细胞雌二醇的分泌均有促进作用,其中 F_4,F_3,F_2比 F_1更敏感;③加前体物(孕酮或雄烯二酮)之后,再加鸡 GnRH-Ⅱ比单加前体物或单加鸡 GnRH-Ⅱ,膜细胞雌二醇的分泌量增加更明显.实验结果提示,在体外细胞培养的条件下,GnRH-Ⅱ对膜细胞雌二醇的分泌不仅有促进作用,还可能促进雌二醇的合成。     

牛腔前卵泡的简易机械分离方法

采用两种机械法分离牛腔前卵泡。方法一 (M -1) ,皮肤移植刀切割、剪碎、过滤镜下直接捡卵法。方法二 (M -2 ) ,皮质片剪碎、离心、悬浮、过滤镜下捡卵法。M -1平均每卵巢采卵数为 46 9个± 18 2个 ,M -2为 6 8 4个± 12 5个 ;处理时间M -1为 2 5 4min± 6 7min ,M -2为 46 3min± 16 8min ,两种方法采集腔前卵泡数量及处理时间均存在显著差异。平均每小时采集卵泡数分别为 99 5和 87 2个 ,M -1多于M -2。两种方法获得腔前卵泡大小分布规律也有明显差异 ,M -1采集腔前卵泡直径为 6 0～ 15 0 μm ,绝大部分是次级卵泡 ,而M -2获取腔前卵泡则偏小。总的看来 ,M -1处理时间短 ,回收效率高 ,方法简便 ,且分离卵泡直径较大 ,适于体外培养 ,是机械分离牛腔前卵泡的一种理想方法。     

Influence of organic phosphorus on reproductive performance and metabolic profiles of anoestrous Farafra ewes in subtropics at the end of breeding season

The effect of organic phosphorus on metabolic, haematological and hormonal status, restoration of ovarian functions and conception rate in anoestrous Farafra ewes in subtropics were evaluated. Anoestrous Farafra ewes (n  = 24; 34.72 ± 0.52 kg body weight) were allocated into two equal groups: control and phosphorus groups. The ewes of phosphorus group were treated with sodium 4‐dimethylamino‐2‐methyl‐phenyl‐phosphonate as an organic bound phosphorous twice a week for successive 3 weeks. Ovarian follicle development and corpora lutea were checked three times a week till occurrence of oestrus using ultrasonography while pregnancy was confirmed at 30 days post‐service. Plasma metabolites, reproductive hormones, thyroid hormones and minerals were detected at weeks ?2, ?1, 0 (mating day) and + 4 weeks post‐oestrus. Phosphorus group had significantly (p  < .05) short interval to oestrous resumption if compared to control ewes (2.1 ± 0.8 weeks vs . 4.6 ± 1.1 weeks). In addition, phosphorous supplementation significantly (p  < .05) increased the number of antral follicles (developed and their sizes in addition to sizes of corpora lutea (8.72 ± 0.3 mm vs . 7.46 ± 0.9 mm) as well. Number of services per conception (2.6 vs . 1.4; p  < .01) was higher in control group than that of phosphorus group. Pregnancy rate (80 vs . 50%) was significantly (p  < .01) higher in phosphorus group when compared to control. White blood cells in treated ewes (10.8 ± 0.44; p  < .05) and monocytes (2.93 ± 0.13; p  < .01) were higher than that of control group (white blood cells; 9.53 ± 0.50 and monocytes; 2.24 ± 0.14). Metabolic parameters did not differ between phosphorus and control groups during different times of treatment. It could be concluded that phosphorous administration to anoestrous Farafra ewes in subtropics could improve reproductive performance and restore ovarian activity at the end of spring and early summer.     

NPFFR1基因对鹅卵泡颗粒细胞激素分泌和细胞凋亡的影响研究

神经相关肽受体（RFamide-related peptide receptor,NPFFR1）是促性腺激素抑制激素的主要亲和受体,它在调控动物繁殖方面起着重要作用。为了解NPFFR1对鹅卵巢卵泡发育的作用,本研究以42周龄健康产蛋四川白鹅为试验材料（n=9）,利用RT-qPCR法检测NPFFR1基因在等级前和等级卵泡颗粒细胞中的mRNA表达规律;在颗粒细胞中过表达NPFFR1基因,酶联免疫吸附法检测颗粒细胞上清液（n=9）中雌二醇（estradiol,E2）、孕酮（progesterone,P4）和抗缪勒管激素（anti-Mullerian hormone,AMH）的浓度变化,剩余贴壁细胞作一步法TUNEL检测细胞凋亡情况;转录组测序方法筛选大黄卵泡（8~10 mm）颗粒细胞过表达NPFFR1前后表达差异显著基因,并对差异表达基因进行功能聚类分析。结果显示,除F1等级外,其余等级卵泡颗粒细胞NPFFR1表达量均极显著高于等级前卵泡（P<0.01）;过表达NPFFR1后,等级颗粒细胞上清液中的E2和等级前颗粒细胞上清液AMH的含量显著（P<0.05）降低,但孕酮P4含量变化不显著（P>0.05）;转录组测序共筛选到267个差异表达基因（119个下调,148个上调）,这些基因主要富集在生物节律过程、繁殖进程等生物学过程中;同时,与对照组相比,差异基因AMH显著下调表达（P<0.05）,Clock（clock circadian regulator）、FOS（proto-oncogene,AP-1 trans-cription factor subunit）、Per（period circadian regulator）和ANTXR2（cell adhesion molecule 2）分别极显著（P<0.01）或显著（P<0.05）上调表达。上述试验结果提示,NPFFR1可从激素、细胞凋亡和生物节律等多个环节影响卵泡颗粒细胞,参与调控卵泡的时序等级发育。     

眼柄切除及注射黄体酮对中华绒螯蟹幼蟹卵巢发育的影响

采用切除单侧、双侧眼柄及注射黄体酮药物的方法研究对幼蟹卵巢发育的影响。结果表明,切除双侧眼柄可明显增加幼蟹体重、蜕壳频率,极显著提高卵巢指数、肝胰腺指数和卵巢／肝胰腺质量比,极显著增大卵母细胞直径;切除单侧眼柄可极显著增加卵巢指数和卵巢／肝胰腺质量比,极显著增大卵母细胞直径;注射黄体酮药物可极显著增大卵母细胞直径,卵巢指数、卵巢／肝胰腺质量比增加未达统计学上的显著水平。     

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.     

大连地区方氏云鳚卵巢发育的组织学特征及其分期

本文根据1985～1987年在大连地区捕获的方氏云鳚为材料,对其卵母细胞发育各阶段的形态特征进行了描述和探讨。并对方氏云鳚产卵类型及卵子形成过程中生长环的形成等问题进行了阐述。     

Cepharanthine induces autophagy via PI3K/AKT/mTOR signaling pathway in ovarian cancer SKOV3 cells

AIM: To investigate the autophagy of human ovarian cancer SKOV3 cells induced by cepharanthine and to explore its mechanism. METHODS: The effect of cepharanthine on the viability of ovarian cancer SKOV3 cells was measured by CCK-8 assay. The SKOV3 cells were treated with cepharanthine, and then the formation of autophagosome was observed with acridine orange staining under fluorescence microscope. The protein levels of LC3, AKT, p-AKT, mTOR, p-mTOR and GAPDH in the SKOV3 cells treated with cepharanthine were determined by Western blot.RESULTS: Cepharanthine significantly inhibited the viability of ovarian cancer SKOV3 cells in a dose-dependent manner (P<0.05). The number of the intracellular acidic autophagosomes with bright red fluorescence was significantly increased after cepharanthine treatment in the SKOV3 cells. The expression of LC3-Ⅱ in SKOV3 cells was significantly enhanced after cepharanthine treatment. Furthermore, treatment with cepharanthine in the SKOV3 cells also resulted in a significant down-regulation of phosphorylated form of AKT and mTOR (P<0.01), while the total protein level was not changed. Combination of cepharanthine and 3-methyladenine resulted in a substantial decrease in the cell viability compared with using cepharanthine alone.CONCLUSION: Cepharanthine significantly inhibits the growth of human ovarian cancer SKOV3 cells and induces the autophagy, which may be correlated with down-regulation of PI3K/AKT/mTOR signaling pathway.