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1.
The yak (Bos grunniens) is the most important livestock animal in high-altitude regions owing to its prominent adaptability to cold conditions, nutritional deficiencies and hypoxia. The reproductive organs exhibit different histological appearances and physiological processes at different reproductive stages. Hypoxia-inducible factor-1 alpha (HIF-1α) is the regulatory subunit of HIF-1 that crucially regulates the response to hypoxia in mammalian organisms. The goal of our study was to investigate the expression and distribution of HIF-1α in the primary yak reproductive organs at different reproductive stages. Samples of the ovary, oviduct and uterus of 15 adult female yaks were collected and used in the experiment. The expression and localization of HIF-1α proteins and mRNA were investigated using quantitative real-time polymerase chain reaction (qRT-PCR), Western blot (WB) and immunohistochemistry (IHC). The results indicated that the expression of HIF-1α protein in the ovary was higher during the luteal phase than during the follicular phase and gestation period (p < .05). In the oviduct, HIF-1α protein was also more highly expressed during the luteal phase than during the follicular phase and gestation period (p < .01). However, in the uterus, the HIF-1α protein had stronger expression during the gestation period than during the follicular phase (p < .01) and luteal phase (p < .05). The expression of HIF-1α mRNA was similar to that of its protein. Immunohistochemical analysis revealed intense immunostaining of HIF-1α proteins in the follicular granulosa cells, granular luteal cells, villous epithelial cells of the oviduct, endometrial glandular epithelium and luminal epithelium, foetal villous trophoblast, and epithelia of caruncular crypts. This study showed that the expression of HIF-1α in the ovary, oviduct and uterus varies according to the stage of the reproductive cycle. This implies that HIF-1α plays an important role in regulating the stage-specific physiological function of yak reproductive organs under hypoxic environments.  相似文献   

2.
This study was carried out to compare the lectin-binding pattern in the normal and pathological oviduct of sows during the ovarian cycle. Lectin-binding patterns showed differences between segments, phases of ovarian cycle and presence of morphologic lesions. In the infundibulum, it was observed that the cysts, in the follicular phase, reduced Ricinus communis-I (RCA-I) and Ulex europaeus-I (UEA-I) binding. Furthermore, in the pathological oviducts of the luteal-phase group, there is a reduction of Concanavalia ensiformis (Con-A) reactivity in this segment of the tube having wall cysts, adenomyosis and diverticulus. The Arachis hypogaea (PNA) binding in the infundibulum, during the luteal phase, decreased in the tube having adenomyosis. In animals with wall cysts, the Con-A, Triticum vulgaris (WGA) and RCA-I reactivity was minor in the glycocalyx of the isthmus epithelium during follicular phase. Con-A and Dolichos biflorus (DBA) binding pattern was minor in the luteal-phase isthmus of the tube having wall cysts, adenomyosis and diverticulus. In the ampulla, the wall cysts impaired the Con-A reaction only in the basal region of the epithelium, in the follicular phase. Binding with Con-A was decreased in the ampulla of animals in the luteal phase in the tube lesions with cysts and diverticulus. In addition, the diverticulus observed in the ampulla, during the luteal phase, reduced the PNA tubaric binding. The results of this study showed that the morphologic alterations modify the sugar pattern in the oviduct of sows. These modifications in glycoconjugates may be one of the reasons for the failure of fertility in sows.  相似文献   

3.
Bcl-2 and Bax proteins localised mainly in granulosa cells. Primordial and primary follicles of new born rat ovary showed an intensive nuclear staining for Bax but faint staining for Bcl-2. In terms of staining intensity, no remarkable difference was observed within the same stage of developing follicle. Compared to new born rats, granulosa cells of adult and one month old rat ovary showed an increased staining both for Bcl-2 and Bax proteins. No staining was observed in primordial follicles of one month old and adult rats. However, granulosa cells of primary follicles, granulosa cells and theca cells in tertiary follicles of adult rat ovary also showed a strong staining for Bcl-2 and Bax proteins. Oocytes of follicles from different developmental stages revealed an apparent staining both for Bcl-2 and Bax proteins. However, in the more mature follicles oocytes stained more intensively. In developing corpus luteum a remarkable staining was observed for Bax. However, the staining was more prominent in regressing corpus luteum. Contrary to this, Bcl-2 stained the luteal cells in developing corpus luteum strongly, while in the fully developed corpus luteum no staining for Bcl-2 was observed. In conclusion, there was an apparent relation between the expression of the apoptosis regulating protein Bcl-2 and Bax and follicular development. Thus, during the follicular development Bcl-2 and Bax may be involved in granulosa cell demise in rat ovary. Furthermore, increased levels of Bax and decreased levels of Bcl-2 in the fully developed corpus luteum suggest that Bax plays a role in apoptosis of luteal cells in rat ovary.  相似文献   

4.
The purpose of this study was to investigate the morphological changes in the epithelium of Thai swamp buffalo oviducts at the follicular and luteal phases by histological technique and scanning electron microscopy. The samples from the infundibulum, ampulla, isthmus and uterotubal junction (UTJ) of the oviduct were taken immediately after slaughter at the local abattoir. Noticeable cyclic changes were observed on the epithelial surface of the infundibulum and ampulla, but few changes were present in the isthmus and UTJ. At the follicular phase, the epithelium of infundibulum and ampulla were densely covered with ciliated cells whose cilia concealed the apical processes of the secretory cells. In contrast, the secretory cells dominated in the epithelium at the luteal phase and most of the ciliated cells were hidden by the bulbous processes of these cells. In the isthmus and UTJ at the follicular and luteal phases, the secretory cells were almost flat or gently rounded and covered with numerous microvilli at their apical surface. In conclusion, the histological and ultrastructural observation of Thai swamp oviduct epithelium revealed marked cyclic changes in the cellular differences associated with the main functions of segmental variations.  相似文献   

5.
旨在探究NGF基因的生物学特性及其在母牦牛生殖器官中的表达特性.本研究收集黄体期母牦牛的心、肝、脾、肺、肾以及胎牛期、卵泡期、黄体期、妊娠期母牦牛的卵巢、子宫和输卵管(n=3),利用RT-PCR克隆牦牛NGF基因,并对其序列进行生物信息学分析,利用RT-qPCR技术分析NGF基因的组织表达特性,利用免疫组化技术(IHC...  相似文献   

6.
为了解水牛有腔卵泡闭锁的起因和有腔卵泡中是否存在卵泡颗粒细胞凋亡,本研究选择青春期前水牛和成年(发情间期和发情期)水牛的卵巢,采用石蜡组织切片、HE染色技术进行光镜观察,并采用DNA原位末端标记(TUNEL)技术和透射电镜观察研究了卵泡颗粒细胞凋亡的特征。结果表明:青春期前水牛的健康有腔卵泡上无或仅有极个别凋亡的卵泡颗粒细胞(GCs),而闭锁有腔卵泡上存在有多量凋亡的GCs,凋亡的形式:GCs层内出现单个或多个凋亡细胞,卵泡腔内出现凋亡小体群。在成年水牛发情间期的闭锁大卵泡上,GCs层弯曲皱折,存在多量凋亡的GCs。发情期水牛成熟卵泡的GCs层也存在GCs凋亡,排卵前的GCs层细胞全部脱落到卵泡腔内。本研究首次在同一组织切片上先后分别应用HE染色和TUNEL检查细胞凋亡,且证实了水牛有腔卵泡GCs凋亡的存在。超微结构显示了GCs凋亡的核边集化、核浓缩及凋亡小体的形态。上述结果提示:在水牛闭锁有腔卵泡中存在着GCs凋亡,GCs凋亡是启动水牛有腔卵泡闭锁的潜在机理。  相似文献   

7.
旨在研究RBM3在牦牛(Bos grunniens)生殖相关调控中的作用。本研究以卵泡期、黄体期和妊娠期4~6岁健康雌性牦牛各3头为采集对象,采集其卵巢、子宫和输卵管共9组试验样品,每组设置3个生物学重复。利用基因克隆技术克隆牦牛RBM3基因并进行生物信息学分析;利用实时荧光定量PCR (quantitative real-time PCR,qRT-PCR)和蛋白免疫印迹(Western blot,WB)分别在基因层面和蛋白层面检测RBM3在牦牛子宫、卵巢和输卵管中的相对表达量;利用免疫组织化学(immunocytochemistry,IHC)方法检测RBM3蛋白在试验样品中的分布定位。牦牛RBM3基因(GenBank No.:MF142258.1)被成功克隆并预测出二、三级结构,发现其与野牦牛亲缘性最近,基因编码区第51位、98位核苷酸不同于常见哺乳动物,生物信息学分析预测其编码的蛋白质为稳定的非跨膜蛋白。RBM3蛋白在本试验9组样品的表达量有如下差异:在卵巢中,黄体期显著高于卵泡期和妊娠期(P<0.05);输卵管中,卵泡期显著低于黄体期和妊娠期(P<0.05);子宫上,卵泡期显著高于黄体期和妊娠期(P<0.05)。免疫组织化学结果显示,RBM3在卵巢中的主要表达位置是卵泡膜、颗粒层和黄体细胞;在输卵管的表达位置是黏膜上皮;子宫上的表达主要以子宫内膜和子宫腺为主。本研究结果提示,RBM3可能参与牦牛个体发情周期和妊娠过程的调控以及妊娠识别等过程,为RBM3这一冷应激调节因子参与牦牛对高原环境的适应性生理机制方面的研究提供参考。  相似文献   

8.
试验旨在分析Bcl-2与Bax基因的序列特性,并分析其在母牦牛生殖轴上的表达特点,为探讨其在牦牛繁殖活动中的调控作用奠定基础。试验采集健康母牦牛与母黄牛下丘脑、垂体、卵巢、输卵管及子宫组织样品,通过RT-PCR扩增并克隆Bcl-2与Bax基因,并采用生物信息学软件进行序列分析;利用实时荧光定量PCR法检测Bcl-2与Bax基因在牦牛与黄牛不同组织中的表达差异。结果表明,牦牛Bcl-2编码区全长690 bp,编码229个氨基酸;与黄牛Bcl-2基因核苷酸序列同源性最高,为99.86%,其次是山羊、绵羊,同源性分别为98.41%、97.97%;系统进化树表明,牦牛与黄牛亲缘关系最近。牦牛Bax基因编码区全长579 bp,编码192个氨基酸,与黄牛、藏山羊和金堂黑山羊同源性较高,分别为99.83%、99.48%和99.48%,其次是绵羊、马、人,同源性分别为99.14%、95.34%、94.30%;系统进化树表明,牦牛与黄牛亲缘关系最近。Bcl-2和Bax蛋白不存在信号肽,均为酸性不稳定的疏水蛋白。Bcl-2与Bax基因在黄牛及牦牛下丘脑、垂体、卵巢、输卵管和子宫组织中均有表达,其中牦牛卵巢、子宫中Bcl-2基因表达量分别显著和极显著高于黄牛(P<0.05;P<0.01);牦牛子宫、输卵管中Bax基因表达量显著高于黄牛(P<0.05),牦牛卵巢中Bcl-2/Bax比值极显著高于黄牛(P<0.01),子宫和垂体中显著高于黄牛(P<0.05)。表明Bcl-2与Bax在动物进化中非常保守且在繁殖活动中起重要作用,牦牛卵巢、子宫、输卵管和垂体中的高表达量可能与牦牛处于极端恶劣环境的细胞抗凋亡作用有关。  相似文献   

9.
10.
The purpose of this study was to determine by immunohistochemistry the expression of estrogen and progesterone receptors in ovarian follicular structures from cows with cystic ovarian disease (COD) and to compare these with normal ovarian structures. Secondary, tertiary, atretic, and cystic follicles were evaluated. The follicular cysts of animals with COD presented a significantly higher expression of estrogen receptor alpha in all follicular layers than secondary, tertiary, and atretic follicles in both groups (P < .05). The intensity of estrogen receptor beta in the granulosa cell layer was stronger in tertiary than in secondary and atretic follicles in normal animals (P < .05) and in growing and cystic follicles in animals with COD (P < .05). Theca cells were scarcely stained in the 2 groups. Growing follicles and cysts from COD animals were less stained than tertiary follicles from normal animals (P < .05). Differences did not exist between the 2 groups with regard to the progesterone receptor. Ovaries of animals with COD exhibited altered estrogen receptors expression compared with that in normal animals.  相似文献   

11.
Leptin is mainly synthesised in white adipose tissue. Besides its effects on body weight and metabolic homeostasis, leptin also has effects on puberty, sexual maturation and reproduction. In this study the relationship between leptin, IGF-1, oestradiol (E2) and progesterone levels were investigated in serum and follicular fluid from cows. This study included 72 healthy, Brown Swiss cows aged 4-5 years. Samples from the jugular vein and follicular fluids were collected. Phases of the oestrus cycle of cows were classified according to their serum progesterone levels (< 3.18 nmol/l, follicular phase and the others as luteal phase). Follicles were grouped as large (> or = 8 mm) or small (< 8 mm). Leptin, IGF-1, oestradiol and progesterone levels were measured from serum and follicular fluid. Leptin concentrations were found to be significantly higher in luteal-phase follicular fluid of small follicles (P < 0.05). These were classified as atretic follicles. There was a positive correlation between serum and follicular fluid leptin levels in the luteal phase. Serum leptin was found to have a positive correlation with follicular fluid progesterone level (P = 0.01) in the preovulatory follicles. The present study shows that there is a relationship between the concentration of leptin in follicular fluid and atresia in small follicles.  相似文献   

12.
旨在克隆获得牦牛StAR基因编码序列(CDS)并进行生物信息学分析,探究其mRNA组织表达特性。本研究以屠宰场采集的成年母牦牛心、肝、脾、肺、肾、卵巢、输卵管、子宫组织(n=5),不同年龄(胎牛、1岁、2岁)牦牛的卵巢(n=3),不同发情周期(卵泡期、黄体期)的牦牛卵巢(n=3),黄体期黄牛的卵巢(n=3)及实验室冻存的牦牛颗粒细胞为研究材料。以牦牛黄体期卵巢cDNA为模板,用逆转录PCR克隆StAR基因,并使用MEGA7.0和ExPASy-ProtParam等软件分析其生物信息学特性;采用实时荧光定量PCR技术分析牦牛StAR基因组织表达特性。结果发现,StAR基因CDS区长858 bp,编码285个氨基酸,StAR蛋白总体带正电荷,属于碱性亲水稳定蛋白,无跨膜结构及信号肽,主要存在于细胞质和线粒体; StAR基因具有较高的保守性,符合物种进化规律。牦牛StAR基因在卵巢表达水平最高(P<0.01),且2岁时卵巢表达水平极显著高于胎牛和1岁龄(P<0.01),黄体期卵巢表达水平极显著高于卵泡期(P<0.01);黄体期黄牛卵巢中StAR基因的表达量极显著高于牦牛(P<0.01);在颗粒细胞的体外培养过程中StAR基因表达量逐渐上升,在培养24 h时达到高峰(P<0.01),随后显著降低。综上所述,StAR基因序列较为保守,在牦牛卵巢组织中表达最高,且表达水平随年龄与卵巢周期而变化,提示StAR基因可能参与牦牛卵巢及黄体功能相关的繁殖调控。  相似文献   

13.
The present study was designed to examine the influence of gonadotrophins treatment on the ovarian morphology changes and plasma concentrations of steroid hormones in peripheral blood. The experiment was performed on sexually pubertal gilts (Large White x Landrace) of similar age (7-8 months) and body mass (100-110 kg) with two controlled subsequent estrous cycles. The animals were randomly divided into four groups: two control consisting of pigs with the luteal phase (n = 9, the 10th day of the estrous cycle) and the follicular phase (n = 6, the 20th day of the estrous cycle) and two experimental ones consisting of animals with both mentioned periods (n = 7 and n = 9) treated with gonadotrophins (PMSG and hCG). The gilts in the luteal phase were injected (s.c.) with gonadotrophins at a daily dose of PMSG 400 and hCG 200 IU from the 16th to the 27th day (the 6th day of the next estrous cycle). The gilts in the follicular phase, were injected with the same dose of gonadotrophins but from the 8th to the 19th day of the estrous cycle. Plasma concentrations of P4, A4, T, E1, E2 and metabolite of PGF2 alpha-PGFM were determined by radioimmunoassay (RIA) method. Injections of PMSG and hCG in both experimental groups produced several times enlarged: weight, size and volume of ovaries and alterations in a number of structural elements as compared with those found in the control animals. The morphological elements presented in ovaries: corpora haemorrhagica, corpora lutea, regular and atretic follicles and first of all cysts by distinctly differentiation thickness of the walls are characteristic for cystic ovarian degeneration. Plasma concentrations all determined hormones after gonadotrophins treatment in experimental groups were increased except E1 (insignificant decrease) in luteal phase as compared with those found in the control groups. Statistically significant increase (p < 0.001) in plasma concentrations of P4, A4, and T in both experimental groups and E2 (p < 0.001) in luteal phase were noted. In peripheral plasma concentrations increase of E1 and E2 in follicular phase of the estrous cycle were insignificant.  相似文献   

14.
Cystic ovarian disease (COD) is an important cause of infertility in cattle, and ACTH has been involved in regulatory mechanisms related to ovarian function associated with ovulation, steroidogenesis, and luteal function. Here, we examined the localization of 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) and 11βHSD2 proteins in the ovary of healthy cows and animals with spontaneous and ACTH-induced COD and the in vitro response of the follicular wall exposed to ACTH. After stimulation by ACTH, we documented changes in 11βHSD expression and cortisol secretion by the follicular wall of large antral and follicular cysts. Follicular cysts showed a higher constitutive expression of both enzymes, whereas ACTH induced an increase in 11βHSD1 in tertiary follicles and follicular cysts and a decrease in 11βHSD2 in follicular cysts. Moderate expression of 11βHSD1 was observed by immunohistochemistry in granulosa of control animals, with an increase (P < 0.05) from primary to secondary, tertiary, and atretic follicles. The level of immunostaining in theca interna was lower than that in granulosa. The expression of 11βHSD2 was lower in the granulosa of primary follicles than in that of secondary, tertiary, and atretic follicles and was lower in the theca interna than in the granulosa. In ACTH-induced and spontaneously occurring follicular cysts, differences from controls were observed only in the expression of 11βHSD1 in the granulosa, being higher (P < 0.05) than in tertiary follicles. These findings indicate that follicular cysts may be exposed to high local concentrations of active glucocorticoids and indicate a local role for cortisol in COD pathogenesis and in regulatory mechanisms of ovarian function.  相似文献   

15.
16.
Dihydrotestosterone (DHT) induces follicular atresia under experimental conditions. However, whether it causes any antagonistic effect under natural condition is not known. In the present study, we investigated concentrations of DHT in follicular fluid and correlated them with concentrations of estradiol-17beta (E2) and its androgen substrates, androstenedione (A4) and testosterone (T), in healthy and atretic follicles of sheep. Merino ewes were treated twice with PGF2alpha (PG) to synchronize estrus. The ovaries were recovered at 14 days after the second PG (luteal phase) or 24h after the third PG given 14 days after the second PG (follicular phase). Follicles were dissected and their size and appearance were recorded. Follicular fluid was collected from follicles larger than 3.5mm and concentrations of E2, progesterone (P4), A4, T and DHT were determined by RIA. The inhibitory effect of DHT on conversion of T to E2 was tested in cultured granulosa cells. Appreciable levels of DHT were observed in the follicular fluid of ovine preovulatory follicles. The levels of DHT were much lower than those of E2, A4 and T, irrespective of physiological conditions of follicles. No difference was found in DHT concentration between healthy and atretic follicles. Dihydrotestosterone marginally inhibited aromatization of T in granulosa cells but this effect was only observed when the levels of DHT were 10 times higher than that of T in culture medium. These results indicate that DHT is present in ovine preovulatory follicles but its levels are not sufficient to exert any antagonistic effect on follicular development.  相似文献   

17.
A three-dimensional model has not been clearly established for the porcine oviductal mucosa. The oviducts of 12 cyclic sows were examined by Scanning Electron Microscopy to study the structure and nature of the mucosal arrangement of the oviduct. Epithelial cyclic changes were also studied. The oviductal infundibulum is an asymmetric funnel-shaped structure surrounding the ostium, in which a wide and a narrow side can be distinguished. The mucosa is more complex in the narrow side, showing numerous and tortuous longitudinal primary folds, while the mucosa becomes simpler in the wide side. Around the ostium abdominale wide secondary folds form cul-de-sacs, with their opening pointing in ovarian direction. Areas between folds throughout the lumen of the oviduct show a high-degree of complexity. Inter-fold spaces are occupied by a system of irregular grooves and pockets, with the presence of basal crypts in the caudal oviduct. Marked variations were observed in the oviductal epithelium depending on the oviductal segment, basal or apical areas of the folds, and phase of the oestrous cycle. Cyclical changes were observed in the infundibulum and ampulla, so that prominent and numerous ciliated cells lined apical areas of the folds in the follicular phase; whereas secretary cells were predominant throughout all areas of epithelial surface in the luteal phase. The oestrous cycle phase appeared do not affect the epithelial population cells of the caudal segments of the oviduct: ciliated and secretory cells uniformly lined apical and basal areas of the folds. The topography of the oviduct provides a complex system of regulation, which may influence not only the passage of cells, but also movement of fluid within the oviductal canal.  相似文献   

18.
Apoptosis is the cellular mechanism of ovarian follicular atresia. The major downstream effector of this phenomenon in many tissues is caspase-3 but little is known about its role in pig ovarian apoptosis. In the present study, we detected the localization of caspase-3 in parallel with nuclear fragmentation (TUNEL) on healthy and early atretic antral follicles. In healthy antral follicles caspase-3 and TUNEL positivity were occasionally recorded within theca layer. The incidence of DNA fragmentation, as indicated also by the biochemical detection, increased mainly in the granulosa layer of early atretic follicles. Quantitative analysis revealed, besides, that atresia was accompanied by a higher incidence of caspase-3 (57.20 +/- 20.05 versus 3.64 +/- 0.61 positive cells in atretic versus healthy follicles, respectively; P < 0.05), of TUNEL positivity (20.13 +/- 9.33 versus 0.42 +/- 0.12; P < 0.05) and simultaneous immunostaining for caspase-3 and TUNEL (15.02 +/- 6.95 versus 0.31 +/- 0.05; P < 0.05) in the granulosa layer. In detached granulosa cells isolated from the follicular fluid of early atretic follicles a further significantly increase was recorded in the percentage of TUNEL positivity and in the incidence of cells that showed colocalization of caspase-3 activity and DNA fragmentation. Granulosa cells of early atretic follicles exhibited a higher positivity for caspase-3 localized in the cytoplasm and occasionally in the nucleus area of granulosa cells. These results indicate that capsase-3 was involved and precociously activated during the process of atresia. Finally, the progressively higher incidence of TUNEL positivity and of double immunostaining in atretic cells collected within the follicular fluid seems to indicate that proteases activity leads only tardily in a detectable DNA fragmentation.  相似文献   

19.
促性腺激素受体在雌性水牛生殖器官的表达定位研究   总被引:2,自引:1,他引:1  
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。  相似文献   

20.
本研究旨在探讨神经生长因子(nerve growth factor,NGF)在母牛卵泡期及黄体期子宫、卵巢、输卵管3种组织中的表达情况。采用FQ-PCR方法和Western blotting技术对成年母牛卵泡期及黄体期的子宫、卵巢、输卵管中的NGF mRNA和蛋白质表达进行了检测。结果表明,NGF mRNA和蛋白质在卵泡期和黄体期的子宫、卵巢、输卵管中均有表达,且卵泡期子宫中NGF mRNA表达量显著高于卵泡期卵巢和黄体期输卵管中NGF mRNA表达量(P0.05),其它组织间无显著差异。  相似文献   

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