牦牛乏情期和怀孕期卵巢微血管密度的测定

为进一步保护牦牛资源和提高牦牛繁殖能力提供基础资料,本研究对牦牛乏情期和怀孕期卵巢中微血管进行了研究。采用免疫组织化学技术和图象分析方法对牦牛乏情期卵巢、怀孕期卵巢微血管密度进行测定。结果乏情期和怀孕期牦牛卵巢微血管光密度值不同。怀孕期有黄体侧卵巢的黄体处微血管光密度值最大,怀孕期有黄体侧卵巢微血管光密度值次之,乏情期卵巢、怀孕期无黄体侧卵巢和怀孕期有黄体侧卵巢的无黄体处微血管光密度值为较小,且后三者之间差异不显著(P0.05),而怀孕期有黄体侧卵巢的黄体处、怀孕期有黄体侧卵巢的微血管光密度值与其他各组间差异均显著(P0.05)。     

发情周期不同阶段牦牛子宫中黄体生成素受体的表达变化

为研究发情周期不同阶段牦牛子宫中黄体生成索受体(LHR)的定位及表达变化,笔者利用免疫组织化学SP法分别检测发情期、发情后期、间情期和发情前期牦牛子宫中LHR的表达,并进行光密度值分析.结果表明,LHR免疫阳性产物在牦牛子宫腺上皮细胞、基质细胞、血管内皮细胞、血管平滑肌细胞和肌层平滑肌细胞中均有表达;腺上皮细胞、基质细胞和肌层平滑肌细胞中LHR在发情前期和发情期表达最弱,发情后期表达增加,间情期表达最强(P＜0.05);子宫内膜血管平滑肌细胞中LHR的表达在发情期最强,间情期最弱(P＜0.05);血管内皮中LHR在发情期和发情前期表达很强,发情后期和间情期显著下降(P＜0.05).结果表明LHR参与了发情周期不同阶段牦牛子宫功能变化的调控.     

促性腺激素受体在雌性水牛生殖器官的表达定位研究

为研究促性腺激素受体(FSHR、LHR)在广西雌性水牛生殖器官中的分布情况,运用免疫组化SABC法对处于不同发情周期(卵泡期、黄体期)成年水牛的卵巢、子宫、输卵管中FSHR、LHR分别进行染色定位。结果表明,FSHR/LHR阳性细胞在卵巢主要见于卵巢内膜细胞及卵泡颗粒细胞;子宫主要见于子宫内膜上皮细胞和腺体细胞;输卵管主要见于柱状上皮纤毛细胞。其中,随着发情周期不同,FSHR、LHR的表达量也有所差异,卵巢中卵泡期FSHR、LHR的表达量均高于黄体期;子宫中FSHR的表达量卵泡期高于黄体期,LHR的表达量黄体期高于卵泡期;而输卵管并没有显著差异。     

卡拉库尔羊卵巢中促性腺激素受体蛋白的分布

在自然发情情况下,取10只卡拉库尔羊卵巢,采用免疫组化链霉菌抗生物素蛋白-过氧化物酶连结法(SP)方法观察卡拉库尔羊卵巢中促卵泡素受体(FSHR)、促黄体素受体(LHR)分布情况。分别选取连续的5张阳性切片,图像分析。结果显示:FSHR、LHR阳性细胞主要分布于卵泡颗粒细胞、膜细胞和卵母细胞。卡拉库尔羊原始卵泡卵母细胞中便有2种受体阳性细胞分布,在各级卵泡中2种受体阳性细胞数量随卵泡发育水平呈正向增加趋势。     

性腺外FSH受体和LH受体表达研究进展

雌性动物的性腺是卵巢,性腺外的生殖组织器官主要包括:输卵管、子宫颈、子宫肌层等,其中子宫是内分泌器官,除有局部内分泌功能外,还可能对下丘脑-垂体-卵巢轴有调节作用.促卵泡素(FSH)和促黄体素(LH)的生理功能是通过其FSHR、LHR来介导的,其受体在性腺和性腺外均有表达.本文主要对雌性性腺外生殖组织器官上两种受体的表达加以综述.     

初情期母牦牛发情期与乏情期主要生殖激素分泌特点的研究

用ELISA对10头发情期内和5头乏情期内的初情期母牦牛进行24 h内血清促性腺激素释放激素(GnRH)、黄体生成素(LH)、卵泡刺激素(FSH)分泌情况检测并比较体重和解剖观察子宫的发育情况。结果显示:发情期母牦牛体重、子宫重、子宫颈-子宫体长、外阴-子宫体长均高于乏情期母牦牛(P0.05),并且在发情期内发情母牦牛高于未发情母牦牛(P0.05);发情期24 h内初情期母牦牛在GnRH、LH、FSH分泌量明显高于乏情期母牦牛(P0.001);在发情期内发情母牦牛GnRH、LH、FSH的分泌量明显高于未发情母牦牛(P0.001)。对比发情期内未发情母牦牛与乏情期母牦牛,二者分泌量均较低,但是发情期内未发情母牦牛激素含量较高并且有较为明显的峰值出现。     

犬不同生殖周期阶段卵巢的组织结构观察

为了探讨犬卵巢组织结构和生殖周期阶段的相关性,试验对犬不同生殖周期阶段卵巢的外观形态和组织结构进行观察。结果表明,犬卵泡期、黄体期和乏情期卵巢体积分别为812.63、1081.80和446.03 mm³,黄体期高于卵泡期和乏情期(P＜0.05),卵泡期高于乏情期(P＜0.05);卵泡期、黄体期和乏情期卵巢质量分别为0.89、1.14和0.71 g,卵泡期低于黄体期且高于乏情期,但3者之间不存在显著性差异(P＞0.05);卵泡期卵巢中可见较多次级卵泡和少量成熟卵泡,黄体期卵巢中可见部分次级卵泡和闭锁卵泡,并有大量黄体存在,乏情期卵巢中卵泡类型主要以原始卵泡为主。可见,犬卵巢形态及组织结构与所处生殖周期阶段有关。     

NGF对输卵管黏膜上皮细胞促性腺激素受体表达的影响

本研究以牛输卵管黏膜上皮细胞为研究对象,利用荧光定量PCR方法检测NGF和酪氨酸激酶抑制剂K252α对输卵管粘膜上皮细胞膜促性腺激素受体FSHR和LHRmRNA表达的影响.添加30、60、100μg/L NGF可以显著增加FSHR mRNA的表达,添加30和60 μg/L的NGF同样增加LHR mRNA的表达.添加抑制剂K252α后,LHR和FSHR的表达量均比对照组显著下降.该结果说明NGF可以通过其受体调节输卵管黏膜上皮细胞促性腺激素受体FSHR和LHR的表达.     

免疫组化法检测促黄体素受体在多浪羊和卡拉库尔羊卵巢上的分布

为从形态学角度理解内分泌调节过程,揭示促黄体生成素(luteotropic hormone,LH)对雌性哺乳动物卵巢调节作用机制,试验采用免疫组织化学SP方法研究促黄体生成素受体(luteotropic hormone receptors,LHR)在多浪羊和卡拉库尔羊卵巢组织中的分布与表达,分别选取相邻的5张连续切片,光镜观察、图像分析。结果显示,多浪羊与卡拉库尔羊的LHR阳性物质主要见于膜细胞、卵母细胞、颗粒细胞和血管周围间质,尤其在膜细胞和卵泡颗粒细胞的胞质中分布最多。原始卵泡卵母细胞中便有LHR受体阳性物质分布,在各级卵泡中LHR阳性细胞数量和染色强度随卵泡发育呈正向增加趋势。     

日粮赖氨酸水平对绿壳蛋种鸡生产性能和血清激素及卵巢FSH和LH受体mRNA表达的影响

本试验旨在研究日粮赖氨酸水平对绿壳蛋种鸡生产性能、血清激素及卵巢FSH受体(FSHR)、LH受体(LHR)mRNA表达的影响。试验选用30周龄的新杨父母代绿壳蛋鸡720只,随机分成6组,每组5个重复,每个重复24只鸡,分别饲喂赖氨酸水平为0.60%、0.65%、0.70%、0.75%、0.80%和0.85%的日粮。结果表明:0.65%赖氨酸水平组产蛋率显著高于0.85%水平组(P0.05),0.70%赖氨酸水平组料蛋比显著低于0.60%水平组(P0.05);血清FSH和LH含量均以0.70%赖氨酸水平组最高(P0.05);与0.75%和0.85%赖氨酸水平组相比,0.65%水平组卵巢FSHR mRNA的表达量显著提高(P0.05);血清FSH含量与卵巢FSHR、LHR mRNA表达量呈现较强相关性(r~20.85)。二次曲线分析表明,以产蛋率和料蛋比为主要评价指标,绿壳蛋种鸡赖氨酸需要量分别为0.70%和0.73%。综上所述,日粮赖氨酸水平为0.70%~0.73%时能够提高新杨绿壳蛋种鸡(31~43周龄)生产性能、血清促性腺激素含量及卵巢FSHR基因的表达。     

Immunohistochemical Localization of Luteinizing Hormone Receptor in the Cyclic Gilt Ovary

Luteinizing hormone receptor (LHR) is a specific membrane receptor on the granulosa and theca cells that bind to luteinizing hormone (LH), resulting in androgen and progesterone production. Hence, the regulation of LHR expression is necessary for follicle maturation, ovulation and corpus luteum formation. We examined the immunolocalization of LHR in cyclic gilt ovaries. The ovaries were obtained from 21 gilts aged 326.0 ± 38.7 days and weighing 154.6 ± 15.7 kg. The ovarian tissues were incubated with rabbit anti‐LHR polyclonal antibody. The follicles were categorized as primordial, primary, preantral and antral follicles. Ovarian phase was categorized as either follicular or luteal phases. The immunolocalization of LHR was clearly expressed in primary, preantral and antral follicles. LHR immunostaining was detected in the cytoplasm of granulosa, theca interna and luteal cells. LHR immunostaining was evaluated using imaging software. LHR immunostaining in the theca interna cells in antral follicles was almost twice as intense as that in preantral follicles (65.4% versus 38.3%, P < 0.01). LHR immunostaining was higher in the follicular phase than in the luteal phase (58.6% versus 45.2%, P < 0.05). In conclusion, the expression of LHR in the theca interna cells of antral follicles in the follicular phase was higher than in the luteal phase. The expression of LHR in all types of the follicles indicates that LHR may impact follicular development from the primary follicle stage onwards.     

Protein and mRNA expression of follicle‐stimulating hormone receptor and luteinizing hormone receptor during the oestrus in the yak (Bos grunniens)

Follicle‐stimulating hormone (FSH) and luteinizing hormone (LH) have a central role in follicle growth, maturation and oestrus, but no clear pathway in the seasonal oestrus of yak (Bos grunniens) has been found. To better understand the role of FSH and LH in seasonal oestrus in the yak, six yaks were slaughtered while in oestrus, and the pineal gland, hypothalamus, pituitary gland, and gonads were collected. Using real‐time PCR and immunohistochemical assays, we determined the mRNA and protein expression of the FSH and LH receptors (FSHR and LHR) in these organs. The analysis showed that the FSHR mRNA expression level was higher in the pituitary gland tissue compared with LHR (p < .01) during oestrus. By contrast, there was low expression of FSHR and LHR mRNA in the pineal gland and hypothalamus. FSHR mRNA expression was higher than that of LHR (p < .05) in the ovary, whereas LHR mRNA expression was higher than that of FSHR (p < .01) in the uterus. FSHR and LHR proteins were located in the pinealocyte, synaptic ribbon and synaptic spherules of the pineal gland and that FSH and LH interact via nerve fibres. In the hypothalamus, FSHR and LHR proteins were located in the magnocellular neurons and parvocellular neurons. FSHR and LHR proteins were localized in acidophilic cells and basophilic cells in the pituitary gland, and in surface epithelium, stromal cell and gland epithelium in the uterus. In the ovary, FSHR and LHR protein were present in the ovarian follicle. Thus, we concluded that FSHR and LHR are located in the pineal gland, hypothalamus, pituitary and gonad during oestrus in the yak. However, FSHR was mainly expressed in the pituitary gland and ovaries, whereas LHR was mainly expressed in the pituitary gland and uterus.     

Lectin histochemical pattern on the normal and cystic ovaries of sows

The lectin histochemical pattern (LHP) was characterized and compared in normal and cystic ovaries of sows. Six biotinylated lectins (PNA, SBA, WGA, RCA‐1, DBA and UEA‐1) were used on tissue sections. In the normal ovaries, the reaction to UEA‐1 and SBA was mild to moderate in mesothelial and endothelial cells. RCA‐1 staining was mild to moderate in theca interna of growing follicles, corpora luteum and mesothelium. In addition, this lectin presented strong reaction in endothelial cells, granulosa cells of atretic follicles, zona pellucida of growing follicles and plasma. DBA showed strong intensity in mesothelial and endothelial cells. There was mild to moderate reactivity to WGA in granulosa cells, corpus luteum and theca interna of follicles in development, and moderate in zona pellucida, in granulosa cells of atretic follicles and mesothelium. PNA staining was mild to moderate in oocytes and in the adventitia and media of medullary arteries. Changes in the LHP of the cystic ovaries were noted; however, there were no differences in these findings between the follicular and luteinized cysts. UEA‐1 reactivity in the cystic ovaries was moderately reduced in the mesothelial and endothelial cells, whereas there was mild reduction in the DBA staining in the granulosa cells. Reaction to RCA‐1 and WGA in the cysts also was decreased in theca interna, zona pellucida and granulosa cells of atretic follicles. Furthermore, endothelium and theca interna in the cystic ovaries presented mild reduction of marcation to SBA, whereas there was decreased reactivity to PNA in the oocytes and adventitia and media layers of the medullary arteries. The results of the current study show that cysts modify the LHP in swine ovaries. These changes of glycoconjugates in many ovarian structures could modify diverse process and may be one of the reasons for decreased fertility in sows.     

不同生殖周期阶段对犬卵巢卵母细胞体外发育的影响

试验以屠宰场犬卵巢为材料,研究了不同生殖周期阶段(卵泡期、黄体期和乏情期)对犬卵母细胞体外发育的影响。结果表明,从卵泡期卵巢采集A类COCs的数量为22.67±11.02个,显著高于黄体期10.67±5.51个和乏情期7.25±4.92个(P＜0.05);体外培养48 h时,卵泡期、黄体期和乏情期卵母细胞达到MⅠ-MⅡ期比率分别为19.2%±15.3%、26.8%±21.1%、9.8%±10.4%,卵泡期与黄体期之间差异不显著(P＞0.05),但两者均显著高于乏情期(P＜0.05);体外培养72 h时,卵泡期、黄体期和乏情期卵母细胞达到MⅠ-MⅡ期比率分别为25.2%±13.5%、6.6%±6.7%、9.2%±5.5%,卵泡期显著高于黄体期和乏情期(P＜0.05)。同时,随体外培养时间延长,不同生殖周期阶段犬卵母细胞达到MⅠ-MⅡ期比率存在不同变化。说明不同生殖周期阶段对犬卵巢卵母细胞体外发育有一定影响,且与培养时间有关。