Changes in follicular vascularity during the first follicular wave in lactating cows

Increase in the blood supply to individual follicles appears to be associated with follicular growth rates and the ability to become the dominant follicle, while reduced thecal vascularity appears to be closely associated with follicular atresia. Therefore, this study aimed to determine the real-time changes in the vascularity of the follicle wall during the first follicular wave in cycling Holstein cows. Normally cycling and lactating cows (n=5) were examined by transrectal color Doppler ultrasonography (the sensitivity for velocity: > 2 mm/sec) to determine the changes in the vasculature of the follicle wall (presence or absence of blood flow) and the diameter of follicles. A new follicular wave and ovulation were induced by GnRH injection at 48 h after an injection of PGF2alpha analogue. The ovaries were scanned daily for 7 days after GnRH injection. Follicles >2.5 mm were classified into 3 groups by the changes in diameter as follows: 1) largest follicle, 2) second largest follicle, and 3) small follicles, which included all other follicles >2.5 mm. Before the follicle selection, there was no significant difference in the percentage of follicles with detectable blood flow between the subsequently determined largest and second largest follicles. After the follicle selection, the percentage of follicles with detectable blood flow significantly decreased among the second largest follicles. In addition, small follicles with detectable blood flow kept larger diameters than those without detectable blood flow from one day before the occurrence of follicle selection. It is likely that maintenance of follicle vasculature and appropriate blood supply to the larger follicles is essential for follicle dominance. In small follicles, the presence of blood flow within the wall also appears to be required for recruitment. Consequently, the data suggest that the change of the blood supply to an individual follicle closely relates to the dynamics of follicular growth in the first follicular wave in the cow.     

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.     

Black-hair follicular dysplasia in dogs

Black-hair follicular dysplasia in dogs of mixed breeding was delineated by hypotrichosis and dullness of most black regions of the coat. White regions remained full and lustrous. The abnormality was not grossly visible at birth but became evident after the 1st month of life. Among 15 offspring from 2 matings of an affected pair of dogs, each of 12 spotted pups developed the disorder in black regions; the remaining 3 were all white and appeared normal.     

Gonadotropin concentrations, follicular development, and luteal function in pituitary stalk-transfected ewes treated with bovine follicular fluid.

Two experiments, each arranged as a 2 x 2 factorial, were conducted in ewes to examine direct effects of bovine follicular fluid (bFF) on follicular development and luteal function and to further characterize follicular development and luteal function after pituitary stalk transection (SS). In Exp. 1, ewes were sham-operated or SS on d 6 of an estrous cycle and received 5 ml of saline or bFF three times daily on d 5 through 11 of the same cycle. In Exp. 2, all ewes were SS on d 6 of an estrous cycle and treated with saline or bFF three times daily on d 5 through 11 and with ovine FSH (60 micrograms; NIADDK-oFSH-16) or saline (1.2 ml) from d 7 to 11. In Exp. 2, ewes were ovariectomized on d 11 to assess effects of treatments on follicular development and luteal function. In both experiments, concentrations (ng/ml) of FSH on d 7 were suppressed (P less than or equal to .005) by bFF compared with saline (.50 +/- .17 vs 1.63 +/- .15) and remained suppressed (P less than or equal to .005) through d 11 (.46 +/- .12 vs 1.54 +/- .12). Replacement therapy (oFSH) restored concentrations of FSH. Concentrations of LH were not affected by bFF but were elevated (P less than or equal to .05) 1 d after SS (d 7; .88 +/- .09 vs .56 +/- .09) and remained elevated (P less than or equal to .05; 1.31 +/- .20 vs .65 +/- .11) from d 6 through 11. Concentrations of progesterone were unaffected by SS.(ABSTRACT TRUNCATED AT 250 WORDS)     

Observations on the size of the follicular hierarchy in the turkey hen and a case of arrested follicular growth

1. The phase of rapid development of ovarian follicles in turkeys fed ad libitum or given a restricted allocation of food from 6 to 30 weeks of age was determined by feeding a fat soluble dye and counting the number of rings of dye in the yolks of laid eggs. 2. The duration of rapid growth was similar in ad libitum and restricted birds and averaged 11.6 d. The maximum number of yellow follicles greater than 8 mm expected in the hierarchy of the turkey hen is 9 to 10 compared with 6 to 7 in the chicken. 3. Discontinuous growth was observed in a series of follicles from one bird. This is believed to be the first report of arrested follicular growth in the turkey.     

Ovine follicular fluid inhibits aromatase activity

“Within follicle” regulations may be important for the fine tuning of gonadotrophin action in ovarian follicles. While numerous growth factors, steroids or proteins which are present in follicular fluid have been shown to have the ability of positively or negatively affecting follicle function, the net effet of follicular fluid of the dominant follicle on its function is unclear.

A bioassay measuring aromatase activity of follicular walls was used (1) to check whether follicular fluid from dominant follicles can alter aromatase activity (2), to check how follicle size, atresia and specific gonadotrophins alter the effects of follicular fluid (3), to identify the nature (steroid or protein) of the active compound(s), and (4) to check whether the inhibition is specific of aromatase. Dominant follicular fluid had the ability to reduce aromatase activity. This effect was dose dependent and was obvious whether or not a protease inhibitor was added to the incubation medium. There was no difference in the magnitude of the inhibitory effect of follicular fluid when FSH (2 ng/ml) or no FSH was added to the incubation medium. LH, however, could potentialise the inhibitory effects of follicular fluid. Dominant follicular fluid was more potent to inhibit aromatase than follicular fluid from atretic follicles. Medium conditioned by granulosa cells, but not by theca cells could inhibit aromatase activity when added to the incubation medium. Charcoal treatment of dominant follicular fluid did not remove its inhibitory potential. Fractionation of dominant follicular fluid by a desalting column demonstrated that the inhibition was related to a compound(s) > 10 kDa. Finally, the effect of dominant follicular fluid on aromatase appears specific of this enzyme as follicular fluid does not affect androgen output by thecal shells or progesterone output by luteal cells.

Further research is required to check whether the activity observed in dominant follicular fluid is related to compounds known to affect aromatase activity (inhibin, mullerian inhibiting substance, heat shock protein 90, superoxyde dismutase) or to another peptide/protein.     

Relationship between ovarian weight and follicular population in heifers

The size of the ovary varies substantially among cattle. This variation may influence the potential of the ovary to produce follicles. In the present study, we examined whether a relationship exists between the weight of the ovary and the number of antral follicles >or=1 mm. Paired ovaries were obtained from Holstein x Japanese Black F1 heifers. Follicles were classified into three size categories (small: 1.0-<5.0 mm, medium: 5.0-<8.5 mm and large: >or=8.5 mm), and the number of follicles in each category was recorded. Large variations in the weight of ovaries and the number of follicles were observed among animals. Significant positive correlations (r>or=0.4, P<0.001) were found between the weight of intact ovaries and the number of follicles in all three categories for the ovary contralateral to CL (OCC) and in the small follicles for the ovary ipsilateral to CL (OIC). Significant positive correlations (r>0.4, P<0.0001) were also observed between the weight of ovaries devoid of CL and follicles and the number of small and medium follicles in both OIC and OCC, indicating that the correlation is not due to the increase in ovarian weight associated with the increase in follicular number. Paired ovaries contained a similar number of small and medium follicles, and significant positive correlations were observed between them (r>0.6, P<0.0001). There were significant positive correlations between the weight of OCC and the number of small and medium follicles in paired ovaries (r>0.4, P<0.0001). These results suggest that 1) the weight of an ovary reflects the potential of the ovary to produce antral follicles, and 2) a rough estimation of follicular population might be possible by using the weight of the ovary contralateral to CL in heifers.     

Ovarian follicular dynamics in Caspian mares

The characteristics of the major follicular waves (primary and secondary) throughout estrous cycle were studied in 7 healthy Caspian mares (age, 4-15 years; weight, 198.6 ± 0.9 kg) during the breeding season. Ovarian follicular dynamics were monitored by using an ultrasound scanner equipped with a 5-MHz, B-mode, linear-array, rectal transducer throughout 2 complete estrous cycles. The diameters of antral follicles (5 mm) were measured, averaging the narrowest and widest dimensions. To detect follicular wave emergence, the diameter profile of the 3 largest follicles per ovary of each mare was determined without considering day-to-day identity of follicles but with maintenance of distinction between left and right ovaries. The primary waves originated on day 6.4 ± 0.81 (ovulation = day 0) when the mean diameter of ovarian follicles was 9.6 ± 1.05 mm. Divergence between the dominant preovulatory follicle and subordinate follicles occurred on day 13.4 ± 0.81, when the dominant follicle was 18.1 ± 2.67 mm in diameter. The intervals from emergence to divergence and from divergence to ovulation were 7 ± 0.68 and 8.7 ± 0.68 days, respectively. Secondary major follicular waves were not observed during this study. In conclusion, only 1 major follicular wave was detected in a Caspian mare, confirming the data previously described in other equine breeds. It is also indicated that the occurrence of 1 major follicular wave per cycle is a more common phenomena in equine species.     

Concurrent follicular dysplasia and interface dermatitis in Boxer dogs

Recurrent or persistent follicular dysplasia and interface dermatitis are described in nine Boxers. Data on age, sex, seasonality of alopecia and histopathological features of the follicular dysplasia in these nine Boxers are comparable with those described in previous reports. The interface dermatitis was characterized by multifocal annular crusted lesions confined to the areas of follicular dysplasia. The inflammatory lesions were neither pruritic nor painful and affected dogs were otherwise healthy. Histopathologically the clinically inflammatory lesions were characterized as an interface dermatitis. Immunohistochemical studies failed to demonstrate immunoglobulins or complement at the basement membrane zone or within blood vessel walls. In dogs with recurrent or persistent disease, the follicular dysplasia and interface dermatitis ran identical, concurrent courses of spontaneous remission and recurrence, or persistence, respectively. One dog with persistent disease was treated successfully with tetracycline and niacinamide for the interface dermatitis, and melatonin for the follicular dysplasia. Although the aetiopathogenesis of this newly described condition and the relationship between the two histological reaction patterns are not known, photoperiod and genetic predisposition appear to play a role.     

牛卵泡卵母细胞的体外成熟和冷冻保存

在简化牛卵母细胞体外成熟的基础上,观察了保护剂、平衡温度、预平衡方法、解冻方法以及冷冻方法对牛卵泡卵母细胞冷冻的影响。结果表明:在体外成熟培养液中添加26.2 mmol/L的NaHCO3和对屠宰场卵巢进行选择更能促进牛卵母细胞的体外成熟;无论是程序冷冻还是玻璃化冷冻,乙二醇(EG)与甘油(GLY)相比更能促进牛卵泡卵母细胞的冷冻效果;在程序冷冻中,冷冻液中添加0.1 mol/L蔗糖比添加0.3mol/L的蔗糖更能促进牛卵泡卵母细胞的冷冻;在玻璃化冷冻中,37℃和25℃的平衡温度比4℃更适合牛卵泡卵母细胞的冷冻;在10%、5%、1%的预平衡浓度之间,5%的预平衡浓度即可达到预平衡的效果;在解冻时,多步脱除保护剂更能保护冷冻的卵母细胞;比较了几种最小样本量(minimum size sample,MSS)玻璃化冷冻方法,解冻成熟培养后的成熟率,拉细毛细玻璃管冷冻法为(41.67±3.19)%,极显著高于未拉细毛细玻璃管冷冻法(glassmicropipette,GMP)的(30.19±1.93)%和固体表面冷冻法(solid surface vitrification,SSV)的(28.33±2.89)%(P<0.01)。     

Thyroid follicular adenocarcinoma in a ferret

A 5-year-old male castrated ferret was presented to the Washington State University College of Veterinary Medicine for evaluation of progressive hair loss and a large, rapidly growing ventral neck mass. The patient had been diagnosed previously with an insulinoma, which was managed medically. Fine-needle aspirates of the neck mass were performed. The cytologic results were most consistent with epithelial neoplasia, likely a carcinoma; thyroid origin was considered likely based on tumor location and cell morphology. The tumor grew rapidly, and the owners elected euthanasia 1 week after examination. At necropsy, a circumscribed, ovoid mass disrupted the right cervical musculature next to the right lobe of the thyroid gland. Histopathologic evaluation revealed an infiltrative mass consisting of cuboidal cells arranged in solid sheets and irregular follicles enclosing colloid. The cells were large, with prominent nucleoli, and had a high mitotic rate. The histopathologic diagnosis was consistent with thyroid follicular adenocarcinoma. Immunochemical findings confirmed thyroglobulin production by neoplastic cells, but to a lesser extent than in normal ferret thyroid tissue. To our knowledge, this is the first case of thyroid follicular adenocarcinoma to be reported in a ferret, with only 1 other case of thyroid carcinoma, a C-cell carcinoma, described previously.     

卵泡抑制素主动和被动免疫及基因免疫研究进展

抑制素是由动物卵巢颗粒细胞或睾丸间质细胞分泌的糖蛋白激素 ,对垂体FSH的分泌具有抑制作用。用抑制素或其片断主动或被动免疫可以提高家畜的排卵率和产仔数 ;用抑制素基因免疫可望得到类似效果。本文通过分析抑制素的理化特性、生物学作用及作用机制、主动和被动免疫对家畜繁殖力的影响 ,阐述了抑制素基因免疫的原理与方法及应用前景     

Effect of charcoal-extracted porcine follicular fluid and 17 beta-estradiol on follicular growth and plasma gonadotropins in gilts fed a progesterone agonist, altrenogest

Thirty-four gilts in two experiments were fed altrenogest for 18 d to block spontaneous growth of ovulatory follicles after luteolysis. They were injected with estradiol or charcoal-extracted porcine follicular fluid (pFF) to determine 1) whether gonadotropin secretion could be depressed and 2) whether exposure to reduced levels of gonadotropins would result in decreased numbers of medium follicles (3 to 6 mm in diameter). Gilts in Exp. 1 received treatments in a 2 X 2 X 2 factorial arrangement starting 48 h before the last feeding of altrenogest. Corn oil or estradiol (2 micrograms/kg body weight), 5 ml of charcoal-extracted porcine serum (pS) or pFF were injected im four times at 8-h intervals and gilts were sacrificed 24 or 96 h after last feeding of altrenogest. In Exp. 2, gilts received one of four treatments consisting of 1) pS, injected iv nine times at 8-h intervals starting 48 h before the last feeding of altrenogest; 2) pFF, with injection protocol the same as for pS; 3) estradiol injected im three times and 4) four times at 8-h intervals starting 0 and 24 h, respectively, before the last feeding of altrenogest. Compared with pS or corn oil, estradiol increased (P less than .001) plasma estrogen and decreased (P less than .05) plasma luteinizing hormone (LH) without a significant effect on plasma follicle stimulating hormone (FSH). Estradiol, compared with corn oil, decreased (P less than .01) the number of medium follicles from 24.8 to 0/gilt and decreased (P less than .05) the weight of ovarian follicular fluid from 4.2 to 2.1 g/gilt at 72 h after the first injection. Five milliliters of pFF had no significant effect on plasma gonadotropins or number of medium follicles. However, 20 ml of pFF, compared with pS, decreased (P less than .05) plasma FSH from 45 ng/ml to 9 ng/ml 32 h after the first injection, had no effect on plasma LH, decreased (P less than .01) the number of medium follicles from 29.2 to 2.2/gilt and decreased (P less than .01) follicular fluid weight from 3.9 to 1.6 g/gilt by 72 h after the first injection. These results indicate that estradiol or a non-steroidal component of follicular origin can decrease secretion of gonadotropins and suppress recruitment of medium follicles in the pig.     

Changes in follicular fluid steroids, insulin-like growth factors (IGF) and IGF-binding protein concentration, and proteolytic activity during equine follicular development

The objective of the present study was to evaluate changes in equine follicular fluid insulin-like growth factor binding protein (IGFBP) proteolytic activity as well as steroid, IGF, and IGFBP concentrations during follicular development in the mare. Mares (n = 14) were classified as either in the follicular phase (n = 8) or luteal phase (n = 6). Follicles (n = 92) were categorized as small (6 to 15 mm; n = 54), medium (16 to 25 mm; n = 23), or large (> 25 mm; n = 15), and follicular fluid was collected. Estradiol and androstenedione levels in follicular fluid were greater (P < 0.05), and IGFBP-3 concentrations tended to be greater (P < 0.10) in large than in small or medium follicles, whereas IGFBP-2, -4, and -5 levels were less (P < 0.05) in large than in small or medium follicles. Estradiol and androstenedione concentrations were negatively correlated (P < 0.01) with IGFBP-2, -4, and -5 but not IGFBP-3 concentrations. To evaluate proteolysis of IGFBP, follicular fluid was incubated with human 125I-labeled IGFBP-2, -3, and -5 and protein separated by 12% SDS-PAGE. Follicular fluid caused little or no proteolysis of 125I-lableled IGFBP-2 or -3, and the small amount of proteolysis of IGFBP-2 and -3 did not differ (P > 0.10) among follicle classes. However, more 125I-labeled IGFBP-5 was cleaved (P < 0.05) by follicular fluid from large follicles collected during the follicular phase than large follicles during the luteal phase, and small or medium follicles from follicular and luteal phase mares indicating that a protease to IGFBP-5 exists in estrogen-dominant equine follicles. This IGFBP-5 protease was inhibited by kallikrein/serine protease and metalloprotease inhibitors. We conclude that the tendency of estrogen-dominant follicles of mares to have greater levels of IGFBP-3 and lesser levels of IGFBP-2 does not appear to be due to differences in proteolysis, whereas changes in IGFBP-5 levels are likely due to changes in activity of a serine protease or metalloprotease. Changes in IGFBP may alter levels of bioavailable IGF that stimulate steroidogenesis and mitogenesis in developing mare follicles.     

Black-hair follicular dysplasia in a Holstein cow

A color-linked hypotrichosis was documented in an adult, black-and-white, Holstein cow. Skin biopsies from black-haired areas showed varying degrees of follicular dysplasia, while the white-haired areas were normal.