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利用产蛋鸡小黄卵泡膜细胞培养模型研究黄体生成素(LH)对等级前发育的小黄卵泡膜细胞增殖的影响以及LH作用的信号转导途径。单层培养的外层膜细胞用LH(0.1~100μg/L)、蛋白激酶A(PKA)激活剂forskolin(FRSK,0.1~10μmol/L)及抑制剂H89(0.01~1μmol/L)或蛋白激酶C(PKC)激活剂佛波酯(PMA,0.1~10nmol/L)及抑制剂H7(0.01~1μmol/L)单独或共同处理,36 h后测定膜细胞增殖的变化。结果显示,LH和FRSK促进小黄卵泡膜细胞的增殖,且LH的促增殖作用可被H89阻断,而PMA无显著的促进作用,且H7不能阻断LH的促增殖作用。由此推断LH的促小黄卵泡外层膜细胞增殖作用主要是通过PKA途径进行的。 相似文献
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选取10只1.5岁成年小尾寒羊母羊,分别取卵泡颗粒细胞并进行培养。设置5个试验组和1个空白对照组,试验组采用不同质量浓度血管内皮生长因子(VEGF)刺激,空白组不添加VEGF。并用MTT检测细胞增殖情况,Western-blotting检测细胞中P-ERK1/2表达水平。MTT比色结果表明,当VEGF质量浓度为5μg/L时对细胞增殖的促进作用最大,且试验组均高于对照组;Western-blotting结果显示,P-ERK1/2表达水平随VEGF的添加而显著增加,并在5μg/L时达到最大值(P〈0.05)。但在10、15、20μg/L质量浓度下其表达量差异不显著(P〉0.05)。这说明不同质量浓度VEGF对颗粒细胞增殖的促进作用各不相同,其中以5μg/L的VEGF效果最好。当颗粒细胞培养至48h时,P-ERK1/2表达水平较24h时略有提高,并在5μg/L VEGF刺激时达到最大值。VEGF可能是通过促进P-ERK1/2的表达进而促进颗粒细胞的增殖。 相似文献
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为探讨孕马血清促性腺激素(PMSG)对2-3月龄性未成熟的小母猪卵巢中卵泡发育,卵泡闭锁以及卵泡颗粒细胞凋亡的影响,就其注射PMSG后24,48,72,96h卵巢中各类卵泡数量,颗粒细胞凋亡比例及外周血浆中类固醇激素水平等进行了研究。结果如下:(1)未注射PMSG前,性未成熟小母猪卵巢表面卵泡总数(10.5个/卵巢)及各类卵泡数(小卵泡9个/卵巢,中等卵泡数1.5个/卵巢)均较少;注射PMSG后24h,小卵泡数(24个/卵巢),中等卵泡数(6.5个/卵巢)和大卵泡数(1.5个/卵巢)与对照组相比均明显增多;至注射PMSG后96h,小卵泡(4.5个/卵巢)和中等卵泡数(1.5个/卵巢)均明显减少,但大卵泡数量仍较多(4.5个/卵巢)。(2)未注射PMSG前,性未成熟小母猪中类卵泡中的颗粒细胞凋亡比例均较高,小卵泡和中等卵泡分别为24.8%和34.4%;注射PMSG事24h,颗粒细胞凋亡比例显著降低(P<0.05),小卵泡和中等卵泡分别为10.7%和13.7%,至PMSG事72h,小卵泡和中等卵泡的颗粒细胞凋亡比例开始显著升高(P<0.05),而大卵泡在出现后颗粒细胞凋亡比例较低且无较大波动。(2)在未注射PMSG前,血清中的雌二醇含量仅为15.3ng/L,孕酮则由于含量大低而未能检测到;在注射PMSG后,雌二醇和孕酮水平均升高,雌二醇在注射PMSG后72h时达最高值59.2ng/L,孕酮在注射PMSG后48h出现最高值1.7 μg/L。以上结果表明,在性未成熟的2-3月龄小母猪,卵泡颗粒细胞凋亡比例很高,导致卵巢表面各类卵泡数量均较少,血清类固醇激素水平也很低,而PMSG可在其尚未建立起自身下丘脑-垂体-性腺轴的激素调节机制情况,显著抑制颗粒细胞亡,从而抑制卵泡的闭锁,促进卵泡的发育,并进而提高血清中类固醇激素的水平。 相似文献
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同源GnRH对无血清培养鸡卵泡颗粒细胞孕酮分泌的作用 总被引:5,自引:1,他引:4
选用产蛋规律的鸡,在一个产蛋序列中,排卵前1~3h剖腹收集各级卵泡,分离颗粒层建立卵泡颗粒细胞无血清单层贴壁培养模型。在此基础上,用不同剂量鸡促性腺激素释放激素(GnRH-Ⅱ)单独处理或与羊LH(oLH)协同处理,并使用GnRH-Antagonist(GA),以观察GnRH-Ⅱ对颗粒细胞孕酮分泌的影响。研究获得了以下结果:(1)GnRH-Ⅱ对鸡不同卵抱(F1、F3、F5)颗粒细胞孕酮分泌均有促进作用,并呈现剂量-反应关系;(2)GnRH-对oLH促鸡卵泡颗粒细胞孕酮的分泌有明显的协同作用;(3)GnRH拮抗物使GnRH-Ⅱ的促卵泡颗粒细胞孕酮分泌作用受到阻断。 相似文献
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鸡Bcl—2基因的克隆及其对卵泡闰细胞凋亡的影响 总被引:5,自引:1,他引:4
将鸡的Bcl-2基因克隆到真核表达载体JLV,构建了重组质粒JLVB。0.2μg/孔JLVB重组质粒经脂质体介导转染卵泡颗粒细胞,应用流式细胞术等方法分析了转染Bcl-2基因后原代细胞和传代细胞的生长与凋亡情况。与对照组相比,转染后的传代细胞分裂速度较快(P<0.01),凋亡比率较低,G2/M+S期细胞比例极显著高于对照组(P<0.01)。这些结果表明,Bcl-2基因具有促进细胞分裂、抑制细胞凋亡、延长细胞寿命的作用,这种作用是直接的。在转染时间为12h、DNA量为0.3μg/孔的条件下,较好的脂质体介导用量的1.2μL/孔。 相似文献
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旨在研究视神经蛋白(neuropsin,OPN5)对鸭颗粒细胞凋亡、增殖及类固醇激素生成的影响。本研究分别对鸭颗粒细胞进行OPN5过表达质粒和OPN5 siRNA处理72 h (n=6),应用EdU细胞增殖检测技术、流式细胞技术、Annexin V-FITC、RT-PCR、Western blot及ELISA技术系统地检测颗粒细胞的增殖、凋亡情况及生殖相关基因的mRNA、蛋白水平及激素水平的变化规律。结果显示,OPN5过表达能促进鸭卵泡颗粒细胞增殖,抑制鸭卵泡颗粒细胞凋亡;能极显著地促进GnRHR、FSHR、LHR的表达 ,并抑制GnIH、GnIHR的表达(P<0.01);能显著或极显著上调StAR、CYP11A1、3β-HSD、CYP17A1、CYP19A1的mRNA水平(P<0.05或P<0.01);显著升高OPN5、3β-HSD及CYP19A1的蛋白表达水平(P<0.05)和极显著升高E2、P4分泌水平(P<0.01),并极显著降低INHβ的水平(P<0.01)。OPN5 siRNA能够显著降低OPN5的表达水平(P<0.01),抑制卵泡颗粒细胞增殖并促进凋亡,极显著下调GnRH、GnRHR、FSHR、LHR(P<0.01),促进GnIH、GnIHR的表达(P<0.01);并极显著抑制StAR、CYP11A1、CYP17A1的表达(P<0.01);显著降低OPN5、3β-HSD及CYP19A1蛋白表达水平(P<0.05)及极显著降低E2、P4的分泌水平(P<0.01),并能极显著升高INHβ的水平(P<0.01)。研究表明,OPN5能促进鸭卵泡颗粒细胞的增殖,抑制其凋亡,促进颗粒细胞中类固醇激素的生成和分泌。 相似文献
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为探究DNA分化抑制因子3(ID3)影响卵泡选择的分子机制,试验观察比较了35周龄与75周龄京红1号蛋鸡卵泡形态、大小及数量差异,采用实时荧光定量PCR检测ID3在不同发育阶段卵泡中的表达,并通过CRISPRi技术与rhBMP2蛋白分别调控鸡小黄卵泡颗粒细胞中ID3的表达,检测卵泡选择相关基因卵泡刺激素受体(FSHR)与类固醇生成急性调节蛋白(StAR)相对表达量。结果显示:35周龄京红1号蛋鸡等级卵泡数目显著高于75周龄蛋鸡(P<0.05);ID3在35周龄京红1号蛋鸡不同发育阶段卵泡中表达差异不显著(P>0.05),但75周龄蛋Fx(F6/7)卵泡中ID3表达显著高于其他卵泡(P<0.05),且75周龄蛋鸡的大白卵泡、小黄卵泡、Fx卵泡、F3卵泡、F1卵泡及卵巢基质中的ID3表达显著高于35周龄蛋鸡(P<0.05);通过CRISPRi抑制ID3表达时,FSHR与StAR的表达会显著增加(P<0.05);10、50、200 ng/mL rhBMP2能显著诱导ID3的表达(P<0.05),FSHR与StAR的表达下降,但差异不显著。研究证实ID3与鸡... 相似文献
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旨在研究Ghrelin对产蛋鸡等级前卵泡发育的调节作用.通过RT-PCR方法检测卵泡Ghrelin受体(GH-SR)的表达情况,采用组织学和免疫组化方法探讨Ghrelin和促卵泡素(FSH)单独或联合处理对鸡等级前卵泡形态学的变化以及层粘连蛋白(LN)和缝隙连接蛋白43(Cx43)的标记率.结果表明,GHSR在颗粒层和膜层上均有表达,其表达量随着卵泡的发育分别至大白卵泡(LWF)和小黄卵泡(SYF)达到最高,随后逐渐降低.悬浮培养的卵泡经Ghrelin和FSH处理24 h后,LWF和SYF的颗粒层和膜层厚度及颗粒细胞密度均显著增加(P<0.05).同时,颗粒细胞中LN和Cx43的标记率显著提高(P<0.05),且以Ghrelin与FSH联合处理组最为显著.结果提示,Ghrelin可通过提高LN和Cx43的表达以增强颗粒细胞的连接功能,促进鸡等级前卵泡颗粒细胞的增殖,并可协同FSH调节卵泡的发育. 相似文献
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根据已发表的Bcl-2基因序列设计3对siRNA(Small interference RNA)序列,构建干扰表达载体,转染体外培养的鹅卵泡颗粒细胞,48h后利用流式细胞术(Flow cytometry,FCM)检测颗粒细胞Bcl-2蛋白表达量、凋亡和增殖情况,收集细胞培养液用放射免疫分析法检测孕激素(P)的分泌水平;此外,还检测了F1~F4卵泡、最小的排卵前卵泡(The smallest preovulatory follicle,SPF)、小黄卵泡(Small yellow follicle,SYF)和闭锁卵泡(Atresic follicle)颗粒细胞Bcl-2蛋白表达和凋亡情况。结果表明:(1)各级卵泡颗粒细胞Bcl-2蛋白的表达存在着差异,正常卵泡颗粒细胞Bcl-2蛋白表达量显著高于闭锁卵泡(P〈0.05),SPF颗粒细胞Bcl-2蛋白表达水平显著高于SYF(P〈0.05);(2)干扰组Bcl-2蛋白表达水平显著低于所有对照组(P〈0.05),而细胞凋亡指数(Apoptosis index,AI)、增殖指数(Proliferation index,PI)和P分泌水平均高于对照组。 相似文献
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魏静 《四川畜牧兽医学院学报》2009,(4):28-32
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。 相似文献
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本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。 相似文献
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以国际标准强毒R株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离MG,并收集感染鸡所产蛋分离MG。结果表明,人工感染48小时后上、下呼吸道及肺已被全面感染,96小时气囊已被感染,120小时输卵管已能分离到MG,卵巢始终分离不到MG。人工感染鸡自144小时便能在其所产蛋中分离出MG。药物治疗能在72小时内消除感染,油乳剂苗则需24天后逐渐降低蛋内MG分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内MG,但以研制的种蛋浸泡剂药浴效果为最好。 相似文献
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REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air. 相似文献
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Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo. 相似文献