影响小鼠转基因原核胚体我发育的若干因素

探讨了显微注射不同浓度的外源DNA以及温度、培养基等对小鼠原核胚体外培养发育能力的影响。显微注射1．5、15、30mg/L衰变加速因子（CD55）、膜反应溶解因子（CD59）基因后，小鼠原核胚发育到囊胚的比率随DNA质量浓度的升高而依次降低为57．7％、36．6％、10．4％，差异检显著（P＜0．01）。注射1．5mg/L外源DNA，原核胚的囊胚率（57．7％）比对照组（54．5％）稍有提高，但差异不显著（P＞0．05）。1．5mg/L为本试验最佳外源DNA注射质量浓度。37℃、CZB建立 或改进的Witten氏培养液，可取得较好的体外培养效果。     

Embryo transfers in Angora and Saanen goats

Sixteen mixed age Angora does were synchronised by progesterone injections and superovulated with either PMSG or FSH in 1982 and six synchronised Angora and seven Saanens were superovulated with FSH in 1983. All donors entire mated and were subjected to egg recovery by a uterine flush conducted during surgery about five days after oestrus. The mean ovulation rate and number of transferable embryos recovered from Angoras treated with PMSG in 1982 was 9.1 and 5.1, respectively, and for FSH treated donors was 15.1 and 11.0. Results for Angoras treated in 1983 were 5.3 and 5.0 and for Saanens were 29.3 and 25.3. Each year recipient feral and Angora does were synchronised, and 329 embryos were transferred surgically to 151 recipients. One hundred and sixteen (77%) of the recipients kidded producing 191 kids (58% embryo survival). Well fed Angora and Saanen donors superovulated with FSH produced 8 and 17 offspring, respectively, in the year of surgery. This rate of reproduction is about 8 times faster than normal and about double that achieved when donors are superovulateb with PMSG.     

Ovulatory Response and Embryo Yield in Jakhrana Goats Following Treatments with PMSG and FSH

Superovulatory response and embryo production efficacy were investigated in adult (age 2–4 years, average body weight: 27–43 kg) cycling Jakhrana goats (n = 15) under semi-arid environmental conditions of India by administering different superovulatory regimens. Goats were reared under semi-intensive system of management in established farm conditions. To synchronize oestrus, a luteolytic dose of carboprost tromethamine (Upjohn, UK) was administered intramuscularly to all does at the dose rate of 5μg per kg body weight in a double dose schedule with an interval of 11 days. For superovulation, 750 IU of PMSG (Folligon, Intervet, Boxmeer, Holland) per goat was administered intramuscularly 24 h before administering a second dose of luteolytic agent in five does (treatment 1). FSH (Sigma, St. Louis, MO, USA) 12.50 IU per goat was administered intramuscularly in a decreasing daily dose schedule (2.50, 2.50; 1.875, 1.875; 1.25, 1.25; 0.625, 0.625) at 12 h intervals over four days, initiated 48 h before administering second dose of carboprost tromethamine in 5 does (treatment 2). FSH (Super-Ov, Ausa Intern, USA) was administered at a uniform dose rate of 8.33 units per goat intramuscularly at 24 h intervals over three consecutive days (total dose was 25 units), initiated 48 h before administering a second dose of carboprost tromethamine in 5 does (treatment 3). To synchronize ovulation in responders, human chorionic gonadotrophin (hCG, Chorulon, Intervet) was injected intramuscularly at a dose rate of 500 IU in each goat on the day of oestrus appearance. Goats were laparotomized 72–82 h following the onset of synchronized oestrus and their genitalia were flushed using a standard collection procedure. Variability (p > 0.05) in superovulatory response (number of established corpora lutea) was observed: FSH (Sigma), 11.8± 2.9; FSH (Super-Ov), 11.6±4.5; PMSG (Intervet), 8.4±2.3. A similar pattern was reflected in mean embryo and transferable embryo recovery, respectively (p > 0.05): FSH (Sigma), 8.0±1.8, 5.2±1.7; FSH (Super-Ov), 6.6±2.4, 5.4±2.4; PMSG, 5.8±1.9, 3.8±2.2. In PMSG-treated does, comparatively more unfertilized ova or retarded embryos were recovered than in FSH-treated does. The superiority of FSH preparations over PMSG was reflected in terms of total and transferable embryo production (p > 0.05). On average, five transferable embryos (excellent and good quality) were recovered per doe treated with FSH of either source. The mean ova/embryo recovery was satisfactory (55–68%). Results indicated that Jakhrana goats can be superovulated for embryo production using FSH of either source to augment productivity.     

Effects of the 6/15 Robertsonian Translocation in Saanen Goats

Contents: The effects of a 6/15 Robertsonian translocation on growth and reproductive performance were investigated. One carrier Saanen buck was compared with 36 non-carrier Saanen sires. No significant effect was observed on daily growth, libido, or siminal characteristics. Fertility rate and prolificacy of the carrier and normal males were 58.3% and 1.53 ± 0.79 (m ± sd) vs 48.9% and 1.36 ± 0.58 respectively (p > 0.05 ).
Kidding rate and prolificacy of 30 normal and 32 heterozygous daughters of this carrier buck were 60% vs 59% (p > 0.05) and 1.47 ± 0.57 vs 1.34 ± 0.48 (p > 0.05) respectively. Normal does always demonstrated a higher prolificacy (p > 0.05) independent of breeding pratices and service rank .
Inhalt: Einfluß der 6/15 Robertsonschen Translokation bei Saanenziegen
Es wurde der Einfluß einer 6/15 Robertsonschen Translokation auf Wachstum und Fortpflanzung sleistung untersucht. Ein Trägertier (Saanenbock) wurde mit 36 Nichtträgern verglichen. Es wurden keine Effekte hinsichtlich täglichen Wachstums, Libido und Spermacharakteristika festgestellt. Befruchtungsprozentsatz und Nachkommenrate des Trägertieres und der normalen Böcke war 58,3% und 1,53 ± 0,79 gegenüber 1,36 ± 0,58 (p ±0,05). Ablammrate und Nachkommensrate von 30 normalen und 32 heterozygoten Töchtern des Trägerbocks betrugen 60% gegenüber 59% (p ± 0.05) und 1,47 ± 0,57 gegenüber 1,34 ± 0,48 (p ± 0,05). Normale Ziegen zeigen höhere Nachkommensraten (p ± 0,05) unabhängig von Bedeckungspraxis und Besamungsfolge .     

Pseudopregnancy in Saanen Goats (Capra hircus) Raised in Northeast Brazil

The prevalence of pseudopregnancy over 44 months was investigated in 23 Saanen goats raised in Northeast Brazil during continuous oestrous cycling (cyclic group) or after synchronization of oestrus (synchronized group). The goats were monitored by ultrasonography and their plasma progesterone profile. The overall prevalence of pseudopregnancy was 30.4% (7/23). In the cyclic group, 28.6% (4/14) of goats showed pseudopregnancy, while in the synchronized group the prevalence was 33.3% (3/9). There was no significant difference between the groups (p>0.05). The mean (+/- SD) length of pseudopregnancy, as shown by the progesterone profile, was 121.6 +/- 33.5 days, ranging from 70 to 155 days. The study defined the prevalence of pseudopregnancy in Saanen goats raised in Northeast Brazil for the first time. This finding identified a major problem for this breed, as without treatment such animals remain unproductive until the spontaneous resolution of the condition. More research seems desirable to ascertain the prevalence of this condition in other breeds in this region of Brazil.     

兔原核胚体外序贯培养

采用添加 10 % FBS的 RPMI16 4 0培养液和 m RPMI16 4 0培养液对兔原核期受精卵进行了体外序贯培养 ,并与添加 10 % FBS的 RD培养液单一培养作了比较。结果显示 :体外培养至 72 h时 ,2个培养组 8-细胞胚率、桑葚胚率和囊胚发育率无显著差异 (P>0 .0 5 ) ,但 RD单一培养组已有 10 .3%出现退化 ,与序贯培养组之间差异显著 (10 .3%比0 ,P<0 .0 5 ) ;体外培养 16 8h,序贯培养组和 RD培养组的贴附率分别为 5 9.7%和 4 1.4 % ,外延生长率分别为 4 3.1%和 2 2 .4 % ,桑葚胚率为 10 0 %和 89.7% ,脱带率为 33.3%和 5 .2 % ,二者之间贴附率、外延生长率差异显著 (P<0 .0 5 ) ,桑葚胚率、脱带率差异极显著 (P<0 .0 1) ;序贯培养 96 h的囊胚细胞数为 (12 4 .6± 6 .36 )个 /枚 ,RD培养同期囊胚细胞数为 (118.2± 5 .2 5 )个 /枚 ,差异显著 (P<0 .0 5 )。结果表明 ,序贯培养能够有效克服兔早期胚胎发育阻滞 ,促进胚胎的正常生长发育 ,提高胚胎质量 ,并能促进胚胎的孵化和附植     

布尔山羊胚胎移植技术的研究与应(简报)

1999年10～11月份,分3批使用阴道栓+FSH超数排卵供体布尔山羊13只.在放入阴道栓的第8～10 d,连续3d递减量肌肉注射FSH(澳大利亚)320 mg.9只供体羊发情、配种、采胚(有效率69.23%).平均采胚数18.11±5.18枚,其中可用胚平均数15.44±6.31枚(可用胚率85.28%).将139枚7日龄可用胚移植受体关中奶山羊89只,妊娠50只,妊娠率56.18%.其中鲜胚移植妊娠率61.11%(44/72),冻胚移植妊娠率41.67%(5/12),二分割胚移妊娠率20%(1/5).50只妊娠受体羊共产羔68只,每只供体羊平均获羔羊7.56只.供体羊采胚后,平均39.9 d发情,配种,全部妊娠产羔,产羔率200%.胚胎移植羔羊性别、初生重、发病率和布尔山羊自繁羔羊无显著差异,P＞O.05.本次布尔山羊移植产生明显的经济效益,技术成熟,可推广应用,逐步产业化.     

波尔山羊胚胎移植技术的研究与应用

对 75只波尔山羊超排处理 ,共获 A、B级胚胎 776枚 ,平均 (10 .35± 6 .4 5 )枚 /只 ,鲜胚移植受体 36 7只 ,移植产羔率达 5 5 .6 %。应用 CIDR和 PMSG同期发情处理受体山羊 5 6 0只 ,5 4 9只达到同期发情 ,32 h内同期率为 98.0 4 %。囊胚和扩张囊胚在发情后 7d的移植产羔率分别为 6 2 .2 2 % (84 /135 )和 5 8.2 6 % (6 7/115 ) ,显著高于桑椹胚 (43.18% )和孵化囊胚 (5 1.72 % ) (P<0 .0 1) ,而囊胚和扩张囊胚间、桑椹胚和孵化囊胚间差异不显著 (P>0 .0 5 )。受体山羊的黄体数量直接影响到移植产羔结果 ,黄体数≥ 2的受体山羊移植产羔率显著高于黄体数为 1的受体山羊移植产羔率 (P<0 .0 1) ,分别为 6 4 .83% (94 /14 5 )和 4 9.5 5 % (110 /2 2 2 )。PMSG注射时间 (在取 CIDR前 2 d或当天注射 )和剂量对受体羊的黄体数和移植产羔率均无显著影响 (P>0 .0 5 )     

波尔、萨能山羊与崇明白山羊杂交对繁殖性能和血清乳酸脱氢酶活性的影响

为了筛选与崇明白山羊杂交的优势组合,选用波尔山羊♂、萨能山羊♂、崇明白山羊♂各两头分别与20头崇明白山羊♀进行交配,研究不同组合对繁殖性能和血清乳酸脱氢酶(LDH)活性的影响。试验结果表明:①就繁殖性能而言,波尔山羊、萨能山羊与崇明白山羊杂交后,体现出一定的杂交优势。其中,波尔山羊的杂交效果比萨能山羊的杂交效果明显,每头母羊胎产羔2头,活羔1.71头,产羔率为200%,羔羊的断奶成活率平均为100%,产活羔数、产羔率、羔羊成活率、断奶活羔数均为三组中最高,是生产中较为理想的杂交父本。②三种成年公羊血清LDH活性由小到大的顺序是波尔、萨能、崇明,以崇明白山羊的活力为最高;在后代羔羊中,波尔与崇明杂交一代公羊的LDH活性高于萨能杂交一代公羊,也高于崇明羊自交后代,可辅助判断出波尔与崇明羊的杂交优势高于萨能羊,可能是较优杂交组合。     

波尔山羊冷冻胚胎移植技术的研究应用

该研究对进口的冷冻波尔山羊胚胎进行移植,并针对性地进行了以下研究解冻温度对冷冻胚胎移植妊娠率的影响;经产母羊与育成母羊胚胎移植后的妊娠率的比较;不同饲养管理条件对胚胎移植后妊娠率的影响.结果如下①解冻温度分别为25℃、32℃时移植后的妊娠率分别为62.50%、63.33%,无显著差异.②经产母羊胚胎移植后的妊娠率(66%)显著高于育成母羊胚胎移植后的妊娠率(55%).③补饲条件下的胚胎移植妊娠率(62.86%)显著高于不补饲条件下的胚胎移植妊娠率(56.67%).     

波尔山羊胚胎移植影响因素的分析

波尔山羊胚胎移植效果受诸多因素的影响，供体状况、药物、季节等因素使供体超排效果产生较大的差异，而受体状况、胚胎质量等对受体的妊娠率产生较大的影响，这些因素阻碍了波尔山羊胚胎移植的产业化应用。本文对以上影响波尔山羊胚胎移植效果的因素进行了分析，以期为提高胚胎移植效率提供依据，从而促进波尔山羊胚胎移植产业化进程。     

胚胎移植供受体山羊发情控制及配比问题的研究

用含１８甲基炔诺酮６０ ｍ ｇ 的阴道海绵栓预处理１１３ 只供体羊 １４～１５ ｄ后注射 Ｆ Ｓ Ｈ 进行超排,３６ ｈ 或４８ ｈ 后撤栓。１９９ 只受体羊用含炔诺酮５０ ｍ ｇ 的阴道栓处理,撤栓时注射或不注射 Ｆ Ｓ Ｈ 进行同期发情。结果表明,供、受体羊发情开始距撤栓的平均间隔时间分别为（３３．９±９９） ｈ 和（４４．３±１６．１） ｈ（ Ｐ＜ ０．０５）,供、受体羊发情持续期长短相似（ Ｐ＞ ０．０５）。撤栓时注射 Ｆ Ｓ Ｈ ２５ Ｉ Ｕ 或 ３３ Ｉ Ｕ 的受体羊,卵巢上形成功能黄体的比率分别为６４．０％ 和 ６０．７％ ,显著高于不注射 Ｆ Ｓ Ｈ的受体羊（４２．２％ , Ｐ＜ ０．０５）。结论认为,用本研究采用的超排及同期发情程序,受体撤栓时间应与供体相同或早于供体羊１２ ｈ;受体羊撤栓时应注射 Ｆ Ｓ Ｈ ２５ Ｉ Ｕ;根据供体羊回收可用胚胎数及同期发情后可用受体数量,供、受体羊的适宜比例为１∶６～１∶８。     

波尔山羊胚胎移植影响因素的分析

波尔山羊胚胎移植效果受诸多因素的影响,供体状况、药物、季节等因素使供体超排效果产生较大的差异,而受体状况、胚胎质量等对受体的妊娠率产生较大的影响,这些因素阻碍了波尔山羊胚胎移植的产业化应用.本文对以上影响波尔山羊胚胎移植效果的因素进行了分析,以期为提高胚胎移植效率提供依据,从而促进波尔山羊胚胎移植产业化进程.     

波尔山羊超数排卵和胚胎冷冻技术的初步研究

在山东省东营市正农高科技农业园养殖场进行了波尔山羊超数排卵、胚胎冷冻试验。结果显示:放CIDR的供体与自然发情相比较,其超排效果无明显差异(P>0.05),说明CIDR是一种行之有效的超排方法;在常规胚胎冷冻过程中,鲜胚质量是影响冻胚成活率的主要因素;采用蔗糖解冻冷冻胚胎时,其浓度0.5mol/L和1.0mol/L均获得较高的解冻胚胎成活率。     

Ghrelin对莎能奶山羊下丘脑GHRH和SS mRNA表达的调节

为探讨ghrelin对奶山羊下丘脑促生长激素释放激素(GHRH)、生长抑素(SS) mRNA表达的影响,10只青春前期莎能母奶山羊随机分为2组,试验组和对照组,每组5只.试验组羊经颈静脉注入一定量的ghrelin(3.0 μg/kg)3 h后宰杀,用RT-PCR方法检测ghrelin对下丘脑GHRH、SS mRNA表达的影响.结果表明试验组羊下丘脑GHRH mRNA的表达显著上调(P＜0.01);下丘脑中SS mRNA的表达也显著升高(P＜0.05).表明ghrelin在下丘脑水平上对GHRH和SS mRNA的表达具有上调作用,提示ghrelin可能通过GHRH途径调节垂体GH的分泌.     

Effect of Pre‐Treatment of Ovine Sperm on Male Pronuclear Formation and Subsequent Embryo Development Following Intracytoplasmic Sperm Injection

The objective of this study was to determine the effects of various methods of sperm pre‐treatment on male pronuclear (MPN) formation and subsequent development of ovine embryos derived from in vitro‐matured oocytes and intracytoplasmic sperm injection (ICSI). The effect of treatment of injected oocytes with dithiothreitol (DTT) on embryo development was also assessed. In Exp. 1, the injected oocytes with non‐treated sperm were activated with three different procedures. The cleavage and blastocyst rates in those activated with DTT was lower (p < 0.05) than those activated with either ionomycin (Io) + 6‐dimethylaminopurine (6‐DMAP) or DTT + I + 6‐DMAP. In Exp. 2, the effects of sperm pre‐incubated with DTT, sodium dodecyl sulphate (SDS) or DTT + SDS as well as two‐time frozen/thawed sperm (without cryoprotectant) on MPN formation and oocyte activation were examined. The non‐treated sperm served as controls. The MPN formation in DTT + SDS group was higher (p < 0.05) than other groups except for freeze–thaw group. No difference in the rate of activated ICSI oocytes was observed among groups. In Exp. 3, the effect of pre‐treatment of sperm on subsequent development of ICSI embryos and blastocyst cell numbers were examined. The rates of cleavage and blastocyst formation as well as the blastocyst cell numbers were similar among the pre‐treated and control groups. In conclusion, pre‐treatment of sperm with DTT + SDS positively affected MPN formation, although the subsequent development capacity of the resulting embryos remained limited. Moreover, DTT was not effective on oocyte activation compared with Io + 6‐DMAP after ICSI.     

青春期前山羊卵母细胞体外生产胚胎的研究进展

青春期前卵母细胞的来源广泛,作为体外胚胎生产的试验材料,国外这方面的研究报道较多。作者就青春期前山羊卵母细胞体外成熟、成熟后细胞质中细胞器的变化情况、体外受精技术和胚胎体外培养等方面研究情况作一综述,供研究者参考。