Anatomicohistological characteristics of the tubular genital organs of the female collared peccary (Tayassu tajacu) from North-eastern Amazon

The present study examines anatomical and histological characteristics of tubular genital organs and its relationships with the reproductive state of 24 wild adult collared peccary (Tayassu tajacu) females. The tunica mucosa of the uterine tube presents a pseudostratified, intermittently ciliated columnar epithelium. The epithelial secretory cells of pregnant females and females in the luteal phase of the oestrous cycle became taller than the ciliated cells and showed abundant apical secretory blebs, whereas secretory cells of females in the follicular phase showed abundant mucous secretory activity (periodic acid-Schiff positive cells). The uterus is composed of two narrow and convoluted uterine horns, separated by the velum uteri, a small uterine body and a long and muscular cervix. The endometrial lining of both uterine horns and body is a monostratified, columnar ciliated epithelium. Pregnant females and females in luteal phase showed a more developed hyperplasia of the endometrial simple tubular glands than females in the follicular phase. The cervix presents interdigitated rows of mucosal prominences that project into the lumen, structures similar to pulvini cervicali, occluding the cervical canal. In pregnant females, the endocervical canal was filled by a viscous cervical secretion. Females in follicular phase presented a thicker vaginal epithelium than pregnant females and females in luteal phase. The present study suggests that the collared peccary female showed different histological features of the uterine tubes, uterus and vagina in accordance with the reproductive state of the females.     

Anatomicohistological Characteristics of the Tubular Genital Organs of the Female Red Brocket Deer (Mazama americana) in the Peruvian Amazon

This study examined the anatomical and histological characteristics of tubular genital organs of 51 adult female red brocket deer in the wild in different reproductive stages, collected by rural hunters in the north‐eastern Peruvian Amazon. The infundibulum was characterized by a large diameter and the presence of a highly folded and ciliated epithelium, and the isthmus has a growing secretor epithelium and a thicker muscular layer. Whereas ciliated cells are more frequent in the infundibulum, epithelial secretory cells showing abundant apical secretory blebs are more frequent in the isthmus. In non‐pregnant females in luteal phase, the endometrium transforms from a proliferative to a secretory type, showing a significant proliferation of endometrial uterine glands. The red brocket deer has four large circular folds in the cervix. The epithelium of the cervix is composed primarily of secretory cells. In pregnant females, the lumen of the endocervical canal is occupied by abundant mucous secretion. All pregnant females had one embryo or fetus, with a fetal sex ratio of 54.0% females to 46.0% males. This species has a cotyledonary, syndesmochorial and partially deciduate placenta, with 6–7 dome‐shaped caruncles per female. The red brocket deer does not present a true cornification of the vaginal epithelial cells, and no vaginal epithelial pattern was determined according the reproductive state of the female.     

Serial Transrectal Ultrasonography for Monitoring the Reproductive Activity of the Asiatic Black Bear (Ursus thibetanus ussuricus)

This study evaluated the structural changes in the reproductive tract of Asiatic black bears using serial transrectal ultrasonography. In addition, the ultrasonographic observations were compared with the results of vaginal cytology and hormonal analyses. The collection of blood for hormonal analysis, vaginal cytology and transrectal ultrasonography was performed in two bears (Bears 1 and 2) from June 2011 to August 2013 without mating and in a third bear (Bear 3) from April to December 2012, allowing natural mating. Serial ultrasonographic observations showed cyclic changes in ovarian structures (e.g. emergence of small follicles, growth and ovulation of dominant follicles and corpus luteum (CL) formation) during the reproductive cycles of the three bears. The diameter of the uterine horns remained similar throughout the reproductive cycle in Bears 1 and 2, and it remained similar from April until October, but an enlargement containing foetuses was observed in Bear 3 in December. The ultrasonographic observations were consistent with the data obtained through vaginal cytology and progesterone analysis during the reproductive cycle. An average of 4.0 (±0.4) dominant follicles was observed during the oestrous stage (May‐August), during which the superficial cells accounted for >90% of the total vaginal cells. In addition, the detection of an average of 2.6 (±0.2) CL was associated with increased plasma progesterone concentrations (3.0 ± 0.4 ng/ml) between June and December (near hibernation). In conclusion, serial transrectal ultrasonography demonstrated yearly oestrous (ovulation) cycles via follicular dynamics and CL formation on ovaries, accordingly with vaginal cytology and hormonal level in the Asiatic black bear.     

Application of ultrasonography in the study of the reproductive system of tropical jennies (shape Equus asinus)

The use of high-frequency (5 MHz) ultrasonography was studied in 11 jennies (7 non-pregnant and 4 pregnant) to characterize the reproductive organs and follicular activities at different stages of reproduction. The result showed close similarity with the mare. The visibility of endometrial folds increased towards ovulation. A positive correlation (p < 0.001; τ = 0.79) was found between the score of the folds and the size of the dominant follicle. The diameter of the uterus and the size of the dominant follicle were significantly correlated (p < 0.001; τ = 0.80). In pregnant jennies, an embryonic vesicle was detectable at 14 days. Follicular growth was characterized by more than one wave. The smallest ovarian follicle was 2 mm and the largest 40 mm. Depending on the reproductive stage, up to 13 follicles were detected per ovary. After monitoring 84 cycles, a mean (±SD) diameter of 34.4 ± 3.6 mm (27.5–40.2 mm) of the preovulatory follicle and 67.85% incidence of single ovulation were found. The mean (±SD) interovulatory interval was 25.7 ± 6 days. This study proved that high-frequency ultrasonography is highly effective in characterizing the reproductive organs and follicular activity of jennies and could be useful in the reproductive management of donkeys.     

Luteal function and follicular growth following follicular aspiration during the peri-luteolysis period in Bos indicus and crossbred cattle

Follicular estradiol triggers luteolysis in cattle. Therefore, the control of follicle growth and steroidogenesis is expected to modulate luteal function and might be used as an anti‐luteolytic strategy to improve embryo survival. Objectives were to evaluate follicular dynamics, plasma concentrations of estradiol and luteal lifespan in Bos indicus and crossbred cows subjected to sequential follicular aspirations. From D13 to D25 of a synchronized cycle (ovulation = D1), Nelore or crossbred, non‐pregnant and non‐lactating cows were submitted to daily ultrasound‐guided aspiration of follicles >6 mm (n = 10) or to sham aspirations (n = 8). Diameter of the largest follicle on the day of luteolysis (7.4 ± 1.0 vs 9.7 ± 1.0 mm; mean ± SEM), number of days in which follicles >6 mm were present (2.3 ± 0.4 vs 4.6 ± 0.5 days) and daily mean diameter of the largest follicle between D15 and D19 (6.4 ± 0.2 vs 8.5 ± 0.3 mm) were smaller (p < 0.01) in the aspirated group compared with the control group, respectively. Aspiration tended to reduce (p < 0.10) plasma estradiol concentrations between D18 and D20 (2.95 ± 0.54 vs 4.30 ± 0.55 pg/ml). The luteal lifespan was similar (p > 0.10) between the groups (19.6 ± 0.4 days), whereas the oestrous cycle was longer (p < 0.01) in the aspirated group (31.4 ± 1.2 vs 21.2 ± 1.3 days). Hyperechogenic structures were present at the sites of aspiration and were associated with increase in concentration of progesterone between luteolysis and oestrus. It is concluded that follicular aspiration extended the oestrous cycle and decreased the average follicular diameter on the peri‐luteolysis period but failed to delay luteolysis.     

雌性牦牛主要生殖器官中DBI的表达与定位分析

为加深对DBI基因功能的探究,揭示其在牦牛生殖生理过程中的作用.本试验采集健康母牦牛(3～6岁)繁殖周期不同阶段(卵泡期、黄体期和妊娠期)的卵巢、输卵管和子宫组织,共分为9组,每组设置3个生物学重复.采用实时荧光定量PCR (qRT-PCR)和蛋白免疫印迹技术(Western blotting,WB)检测牦牛DBI在繁...     

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.     

Ovarian and Hormonal Responses to Follicular Phase Administration of Investigational Metastin/Kisspeptin Analog,TAK‐683, in Goats

This study evaluated the effects of follicular phase administration of TAK‐683, an investigational metastin/kisspeptin analog, on follicular growth, ovulation, luteal function and reproductive hormones in goats. After confirmation of ovulation by transrectal ultrasonography (Day 0), PGF_2α (2 mg/head of dinoprost) was administered intramuscularly on Day 10 to induce luteal regression. At 12 h after PGF_2α administration, intravenous administration of vehicle or 35 nmol (50 μg)/head of TAK‐683 was performed in control (n = 4) and treatment (n = 4) groups, respectively. Blood samples were collected at 6‐h intervals for 96 h and then daily until the detection of subsequent ovulation (second ovulation). After the second ovulation, ultrasound examinations and blood sampling were performed every other day or daily until the subsequent ovulation (third ovulation). Mean concentrations of LH and FSH in the treatment group were significantly higher 6 h after TAK‐683 treatment than those in the control group (12.0 ± 10.7 vs 1.0 ± 0.7 ng/ml for LH, 47.5 ± 28.2 vs 15.1 ± 3.4 ng/ml for FSH, p < 0.05), whereas mean concentrations of oestradiol in the treatment group decreased immediately after treatment (p < 0.05) as compared with the control group. Ovulation tended to be delayed (n = 2) or occurred early (n = 1) in the treatment group as compared with the control group. For the second ovulation, ovulatory follicles in the treatment group were significantly smaller in maximal diameter than in the control group (3.8 ± 0.5 vs 5.4 ± 0.2 mm, p < 0.05, n = 3). Administration of TAK‐683 in the follicular phase stimulates gonadotropin secretion and may have resulted in ovulation of premature follicles in goats.     

Short‐Term Undernutrition Affects Final Development of Ovulatory Follicles in Sheep Synchronized for Ovulation

The objective of this study was to determine, in sheep, the effect of a short‐term undernutrition on growth dynamics and competence of pre‐ovulatory follicles. Synchronization of sexual cycles and induction of ovulation were performed, with progestagens and gonadotrophins, in 14 adult female sheep. Morphological characteristics and developmental competence of ovarian follicles to achieve ovulation were determined by imaging techniques (ultrasonography and laparoscopy) and blood sampling. All the animals ovulated and mean ovulation rates were similar between groups (2.0 ± 0.6 corpora lutea in control ewes and 2.2 ± 0.8 in undernourished sheep). However, nutritional restriction, even during a short period, was related to the presence of large follicles in static growing phase which, despite reaching ovulation, persisted static during the induced follicular phase and evidenced functional alterations as there was no inhibition of the development of subordinate follicles. Thus, this study suggests the existence of deleterious effects from short‐term undernutrition on functionality of pre‐ovulatory follicles, which can compromise fertility.     

Presence of Leptin and Its Receptor in the Hypothalamus,Uterus and Ovaries of Swine Females Culled with Distinct Ovarian Statuses and Parities

Leptin acts on energy metabolism, affecting reproductive functions through activation of its receptors in the brain and in reproductive organs. This study compared the presence of leptin and its receptor (ObR‐b) in hypothalamus neurons, endometrial glands and oocytes of culled swine females across ovarian statuses and parities. Immunohistochemistry was done in samples of uterus, ovaries and hypothalamus from 28 culled females, using polyclonal antibodies antileptin and ObR‐b. Immunolabelling was compared for sows categorized by parity at culling (0, 1, 2–4 and <4) and ovarian status (luteal and follicular phases of the oestrous cycle and with cysts). Immunolabelling for leptin and ObR‐b in neurons and oocytes was weaker in females with cysts (p < 0.05) than in those at the follicular phase. In endometrial glands, leptin immunolabelling was less intense in females with cysts (p < 0.05), but immunolabelling for ObR‐b was similar across ovarian statuses (p > 0.05). In sows culled with 2–4 parities, leptin immunolabelling in neurons and endometrial glands was more intense than in nulliparous females (p < 0.05). In comparison with sows culled at greater parities, ObR‐b immunolabelling for nulliparous females was less intense in endometrial glands and in oocytes (p < 0.05), but more intense in neurons (p < 0.05). Thus, in swine, the presence of leptin and ObR‐b varies across parities and is more intense in the uterus, ovaries and hypothalamus of females that were cycling before culling than in those having cystic ovaries.     

Clinical relevance of pre‐ovulatory follicular temperature in heat‐stressed lactating dairy cows

Temperature gradients in female reproductive tissues seem to influence the success of key processes such as ovulation and fertilization. The objective of this study was to investigate whether pre‐ovulatory follicles are cooler than neighbouring uterine tissue and deep rectal temperatures in lactating dairy cows under heat stress conditions. Temperatures within the pre‐ovulatory follicle, on the uterine adjacent surface and 20 cm deep within rectum, were measured using fine thermistor probes within 45 min after sunrise (dawn). Cows were selected from synchronized groups for fixed‐time insemination during the warm period of the year. Five cows under direct sun radiation and 11 cows in the shade were included in the study. None of the cows in the sun area ovulated within 24 hr, whereas 10 of the 11 cows in the sun area ovulated. Four of the 10 ovulating cows became pregnant. In the ovulating cows, follicular temperatures were 0.74 and 1.54°C significantly cooler than uterine surface and rectal temperatures, respectively, whereas temperatures in the uterine area were 0.80°C significantly cooler than rectal temperatures. No significant differences among temperatures were found in non‐ovulating cows. Follicular size was similar for ovulating and non‐ovulating cows. Environmental temperatures in the shade area were 6.4°C significantly lower than those in the sun area. Results of this study indicate that pre‐ovulatory follicles are cooler than neighbouring uterine tissue and deep rectal temperatures and those temperature gradients were not found in cows suffering ovulation failure.     

Selected Ovarian Ultrasonographic Characteristics During Vernal Transition are Useful to Estimate Time of First Ovulation of the Year

It is important to get mares pregnant as early as possible after vernal transition and thus, identification signs of impending 1st ovulation of the year are warranted. To identify clinical indicators of an approaching first ovulation of the year, mares were teased with a stallion for oestrous detection starting January 3 and subjected to ultrasonographic examination. Day of first appearance of uterus oedema, follicular wall invagination, intrafollicular echogenicity, double contour of the follicle wall, increase in granulosa thickness, follicular wall hyperechogenicity and appearance of pear‐shaped follicles was registered, as well as follicle diameter and number. Seventy per cent of the mares had anovulatory oestrous periods of 4.6 ± 3.6 days, with an interoestroual interval of 12.5 ± 12.2 days. Number of anovulatory oestruses per mare was 2.4 ± 2.3. Uterine oedema occurred in 77% of the mares, 32.4 ± 25.6 days before ovulation. Invagination of the follicular wall appeared in 44.4% of the animals, 24.5 ± 18.4 days before ovulation. Intrafollicular echogenicity was seen in all mares and double contour of the follicle was seen in 77% of the animals. Both last two characteristics appeared 1–72 days before ovulation. Increased thickness of the granulosa occurred in 66% of the mares, 1–19 days before ovulation. Pear‐shaped follicles and follicular wall hyperechogenicity were detected 3 or less days before the first ovulation, in 44.4% and 55.5% of mares, respectively. Mean number of follicles >15 mm decreased at least 16 days before ovulation. We concluded that no isolated characteristic was a reliable indicator. However, increase in granulosa thickness, formation of a pear‐shaped follicle and follicular wall hyperechogenicity, associated with the reduction of the number of follicles >15 mm in diameter to <3, resulted in the first ovulation of the year in 44–67% of the transitional mares, 1–19 days after the characteristics appeared.     

Immunolocalization of angiogenic growth factors in the ovine uterus during the oestrus cycle and in response to Steroids

The vascular changes associated with endometrial maturation in preparation for embryo implantation depend on numerous growth factors, known to regulate key angiogenic events. Primarily, the vascular endothelial growth factor (VEGF) family promotes vascular growth, whilst the angiopoietins maintain blood vessel integrity. The aim was to analyse protein levels of VEGFA ligand and receptors, Angiopoietin‐1 and 2 (ANG1/2) and endothelial cell receptor tyrosine kinase (TIE‐2) in the ovine endometrium in the follicular and luteal phases of the oestrus cycle and in response to ovarian steroids. VEGFA and its receptors were localized in both vascular cells and non‐vascular epithelium (glandular and luminal epithelium) and stroma cells. VEGFA and VEGFR2 proteins were elevated in vascular cells in follicular phase endometrium, compared to luteal phase, most significantly in response to oestradiol. VEGFR1 was expressed by epithelial cells and endothelial cells and was stimulated in response to oestradiol. In contrast, Ang‐1 and Ang‐2 proteins were elevated in luteal phase endometrium compared to follicular phase, and in response to progesterone, evident in vascular smooth muscle cells and glands which surround TIE‐2‐expressing blood vessels. Our findings indicate that VEGFA is stimulated by oestradiol, most predominantly in follicular phase endometrium, and Ang‐1 and 2 are stimulated by progesterone and were increased during the luteal phase of the oestrus cycle, during the time of vascular maturation.     

产蛋期皖西白鹅生殖系统的形态学观察

研究采用大体解剖法、石蜡切片法、HE染色法等对20只产蛋期的皖西白鹅母鹅生殖系统的形态结构和组织结构进行了观察,结果如下:皖西白鹅母鹅的生殖系统包括卵巢和输卵管两部分,仅左侧发育正常,右侧早已退化,卵巢表面被覆生殖上皮,下方是白膜,其实质由皮质和髓质组成,皮质内含有不同发育阶段的卵泡和萎缩卵泡,大卵泡突出了卵巢表面,卵泡无卵泡腔,也无卵泡液,排卵后不形成黄体。输卵管分为漏斗部、蛋白分泌部、峡部、子宫部和阴道部5个部分,各段均由黏膜层、肌层和外膜构成,黏膜上皮有纤毛,固有层内有腺体和淋巴组织,无黏膜肌层,肌层由内环外纵两层平滑肌组成,外膜为浆膜。     

Long‐Term Effect of Deslorelin Implant on Ovarian Pre‐Antral Follicles and Uterine Histology in Female Rats

The objective of the present study was to investigate the inhibitory effects of long‐term deslorelin implant administration on the ovarian and uterine structures of female rats. A total of 16 non‐pregnant female rats were randomly assigned to two groups, each consisting of eight animals. Animals in the implant group (DESL) received subcutaneously (s.c.) a single deslorelin implant (4.7 mg), an analogue of GnRH, while no treatment was applied to the control group (CON). A single adult male rat was introduced into the cages of both the DESL and CON females after 6 weeks of implant administration. After 1 year of implant administration, all animals were killed and follicular structures and volumes of ovaries and uterus were examined using stereological methods. Stereological observations showed that the mean ovarian total volume of the DESL group (0.28 ± 0.07 cm3) was lower than that of the CON group (1.55 ± 0.23 cm3) (p < 0.001). On the other hand, the total number of pre‐antral follicles in the ovaries of DESL (555.32 ± 151.47) females were significantly lower than the control group (1162.96 ± 189.19) (p < 0.001). In the DESL group, the mean volumes of epithelium, endometrium, myometrium and total volume of the uterus were significantly (p < 0.001) lower than in the control groups. In conclusion, these findings indicate that the long‐term deslorelin implant (i) interferes with the normal cyclicity of female rats and (ii) affects the pre‐antral follicle population. Further studies will be required to determine the effects of long‐term deslorelin treatment on the pre‐antral follicle numbers and future fertility in other species.     

IFN-γ在胚泡植入期大鼠卵巢的分布

为研究IFN-γ对妊娠的免疫调控机制,本文采用免疫组化SP法,对大鼠胚泡植入期卵巢组织内IFN-γ的分布进行了研究。结果表明,IFN-γ在不同发育阶段卵泡的颗粒细胞中均有分布,原始卵泡、初级卵泡、次级卵泡和成熟卵泡的卵母细胞呈中等阳性反应,在原始卵泡、初级卵泡及次级卵泡的卵泡内膜细胞呈IFN-γ强阳性反应,成熟卵泡内膜细胞则呈弱阳性反应;在卵巢组织基质细胞、血管内皮细胞、粒性黄体细胞、生殖上皮也出现强阳性反应,卵泡液呈阳性着色。结果表明,妊娠有机体生理剂量的IFN-γ通过影响卵巢功能参与了胚泡的植入,与妊娠的建立和维持密切相关。     

Caspase-9在肉用绵羊主要生殖器官中的分布研究

通过试验研究凋亡因子Caspase-9在肉用绵羊主要生殖器官中的表达,并探讨其生理意义.利用免疫组织化学的方法对Caspase-9在肉用绵羊主要生殖器官中的表达进行研究.研究结果显示,Caspase-9只在细胞质中表达,其在卵巢中主要分布于原始卵泡、初级卵泡及颗粒性黄体细胞,而卵泡膜性黄体细胞中未见表达;黄体期,主要表达于子宫的浅层腺体腺上皮细胞、子宫内膜上皮细胞、子宫颈黏膜固有层的淋巴小结、输卵管黏膜上皮分泌细胞和纤毛细胞、峡部的浆液性腺的腺上皮细胞;卵泡期,子宫和输卵管各个部位阳性反应不明显.在正常生理情况下,Caspase-9参与了肉用绵羊主要生殖器官周期性变化的调控和子宫黏膜免疫,对生殖器官功能的稳定发挥起着重要作用.     

Endometrial Phospholipase A2 Activity During the Oestrous Cycle and Early Pregnancy in Mares

The aim of this study was to determine phospholipase A2 (PLA2) kinetics and activity in the mare’s endometrium during the oestrous cycle and early pregnancy. Phospholipase A2 is responsible for the liberation of arachidonic acid from phospholipids, which is the first limiting step in prostaglandins synthesis. Phospholipase A2 activity was measured using an assay based on the liberation of oleic acid from 1‐palmitoyl‐2‐[¹⁴C] oleoyl phosphatidylcholine. The enzyme was shown to be calcium dependent, to have an optimum pH of 8 and an apparent Michaelis constant of 127 μm . Enzyme activity was low in the endometrium of early luteal phase tissue but increased significantly (p < 0.001) during the late luteal phase (5.39 ± 0.16; 3.48 ± 0.33, 6.85 ± 0.59, and 9.96 thinsp;± thinsp;1.23 thinsp;nmol oleic acid released/mg protein at oestrus, and Days 3, 8 and 14 after ovulation, respectively). The mean PLA2 activity in endometrial tissue from pregnant mares (4.23 ± 0.74) was significantly lower (p < 0.01) than from cyclic animals during late dioestrus (9.96 ± 1.23). The results indicate that PLA2 activity in equine endometrium changes with the stage of the oestrous cycle and thus may be influenced by systemic hormone concentrations. The inhibitory effects of conceptus products on secretion of prostaglandin during early pregnancy were associated with a competitive inhibitor that decreased endometrial PLA2 activity.