In this study, we investigated the number of follicles, oocyte recovery rate and oocyte competence after in vitro maturation according to the size of follicle. And equine oocyte competence after in vitro maturation was investigated in terms of the diameter of follicle with criteria of maturation: nuclear stage after Hoechst staining. The average number of follicles per ovary with middle size (11-20 mm, 2.68) was higher than those of small (5-10 mm, 0.74) and large size follicle (> 21 mm, 1.63), therefore medium follicle (53.1%) had higher proportion than other size of follicles. The average numbers of follicle per ovary was 5.05. The rate of oocyte recovery in small (54.5%) and middle follicle (50%) was higher than that in large follicle (40.9%). After culture for 48 h in Medium 199, 50%, 45.5%, and 44.4% of oocytes from the follicles with diameters of 5-10, 11-20, > 21 mm, respectively reached the metaphase II stage. This is the first report showing number of follicle, oocyte recovery rate according to follicular size, and in vitro oocyte maturation in Jeju mare in Korea. To fulfill in vitro equine embryo production, further studies such as the seasonal effect, in vitro fertilization etc is need.
Ultrasonographic studies were conducted on eight Murrah buffaloes daily from day 6 postpartum (pp) onwards till day 77 pp to monitor changes in the cervix, uterine horn and ovarian follicular growth and development. The mean size of horn and cervix on day six ($9.07{\pm}0.74$ and $8.58{\pm}0.00cm$) decreased significantly to $4.09{\pm}0.09$ and $3.56{\pm}0.08cm$ by day 27 pp, respectively. Follicles in 50% of the buffaloes ovulated within 24 to 54 days pp and the size of the largest follicle on different days increased to more than 5 mm. The remaining 50 percent of animals ovulated after 65 days postpartum. Large size follicles (>8.5 mm) appeared in six out of eight buffaloes between 10 to 30 days pp and five animals had ovulated during early postpartum period. Waves pattern of follicular growth was observed during early postpartum period. Ovulatory follicles growth rate was more than the anovulatory follicles and increase in size was more as compared to the subordinate follicle. Anovulatory follicles persisted for longer period. Mean size of large follicle was more from day 6 to 41 pp and again from 50 to 65 pp in cyclic animals. Second large follicle were large during early postpartum (18days), thereafter, its size was more in acyclic animals. Small follicles population was less in cyclic animals upto day 50 postpartum. Mean medium size follicle growth pattern did not differ in cyclic and acyclic groups. Large size follicle number was more in cyclic group (5/8) during 14 to 20 days postpartum. Presence of large follicles (>8.5 mm) showed initiation of ovarian activity.
In order to study the mechanism of follicular atresia, the follicles of the porcine ovary were isolated according to the presence or absence of the corpus luteum and their size, and then classified to the normal? or atretic?follicle on the morphological observation such as the transparency, the vascularization of follicle, the nuclear phase of oocyte, and the homogeneity of the granulosa cell layer. The viability of granulosa cells was examined. The concentrations of progesterone ($P_4$), testosterone (T), and estradiol-17 beta ($E_2$) in each follicular fluid were estimated by the radioimmunoassay. The viability of granulosa cells in the atretic follicle was much lower than that of the normal one. The concentration of each steroid hormone increased as the follicular size was increased, was not different in quantity between the normal- and the atretic follicle of which diameter was below 3mm, and were much higher in the atretic follicle than those in the normal one of which diameter was above 7mm. The ratio of the concentration of E2 to T in the large atretic follicle valued higher than that in the normal one, but smaller in the small and medium atretic follicle than that in the normal one. The present study suggests that the mechanism of atresia of the large follicle may be different from that of the small and the medium follicle and that the amount of steroid hormones regarded as the one of the criteria for the atretic follicles.
The mammalian skin contains hair follicles, which are epidermal appendages that undergo periodic cycles and exhibit mini-organ features, such as discrete stem cell compartments and different cellular components. Wound-induced hair follicle neogenesis (WIHN) is the remarkable ability to regenerate hair follicles after large-scale wounding and occurs in several adult mammals. WIHN is comparable to embryonic hair follicle development in its processes. Researchers are beginning to identify the stem cells that, in response to wounding, develop into neogenic hair follicles, as well as to understand the functions of immune cells, mesenchymal cells, and several signaling pathways that are essential for this process. WIHN represents a promising therapeutic approach to the reprogramming of cellular states for promoting hair follicle regeneration and preventing scar formation. In the scope of this review, we investigate the contribution of several cell types and molecular mechanisms to WIHN.
The experiment was designed on 42 non pregnant Black Bengal goat. Out of which 18 were subjected to a superovulatory treatment comprising of eCG and hCG for embryo transfer study. The remaining 24 goats received no treatment and served as control for parameter studied as well as recipient for embryo transfer studies. Important biochemical constituents such as acid and alkaline phosphatase, total protein and cholesterol and inorganic phosphorus were estimated in the follicular fluid of control and treated group and the values were separately recorded for small medium and large size follicle. The results indicated a significant effect on acid phosphotase activity due to size of follicle. The value increased progressively from small to medium and from medium to large follicles. Alkaline phosphotase activity showed reverse trend. Alkaline phosphotase decreased progressively as size increased. The concentration of inorganic phosphorus did not reveal any significant difference between the control and treatment groups and also between the different size follicles. The concentration of protein decreased significantly from small to medium and from medium to large, although no difference was observed between the control and treatment groups. The concentration of Cholesterol in the follicular fluid indicated a significant increase from small to medium and to large follicle. Here also no difference was observed due to treatment. Similar in the composition of follicular fluid in the respect of above mentioned constituents indicated no of super ovulatory treatment on follicular fluid composition.
The purpose of this study was to determine the relationship between conception rate and other parameters (body condition score; BCS, progesterone concentrations and follicle size) before estrus induction with CIDR(intravaginal progesterone-releasing controlled internal drug release). The conception rate in cows with < 2.75, 2.75 to 3.25, and 3.25 <, BCS regardless of AI (artificial insemination) time was 46.6%, 63.3%, and 46.6% at CIDR insertion, respectively. The conception rate regardless of BCS was 54.9% in cows inseminated based on detected estrus, and 48.7% in cows inseminated at 72 to 80 hours (timed artificial insemination, TAI) after removal of CIDR. The conception rate regardless of AI time was 40.0% in cows with low progesterone concentrations (less than 1.0 ng/ml), and 56.6% in cows with high progesterone concentrations (more than 1.0 ng/ml) at CIDR injection. The conception rate regardless of progesterone concentrations was 53.8% in cows inseminated based on detected estrus, and 38.0% in cows of TAI after removal of CIDR. The conception rate regardless of AI time was 43.3% in cows with small follicle (less than 5 mm), 53.3% in cows between 5 mm to 10 mm of follicle, and 63.3% in cows with large folliclc (more than 10 mm) at CIDR injection, respectively. The conception rate regardless of follicle size was 58.4% in cows inseminated based on detected estrus, and 45.9% in cows of TAI after removal of CIDR. These results indicated that if the cows with BCS 2.75 to 3.25, active corpus luteum, and/or large dominant follicle (more than 10 mm) are used for estrus induction, the conception rate will be greater.
The ovaries of 178 Holstein heifers or cows (heifer; 41, 1 parity; 72, 2$\leq$ parity; 65) on Day 6 or 7 (Day 0=day of estrus) were examined by transrectal ultrasonography. Diameter of corpus luteum (CL) and large follicle ( $\geq$ 10 mm), and luteal tissue area were determined by ultrasound system with a 5 MHB rectal probe. Blood samples were taken to progesterone analysis. After selection of recipients, frozen Holstein embryos were thawed and directly transferred to recipients non-surgically. The diameter of CL and luteal tissue area was greater (P<0.01) on Day 7 than on Day 6 in heifers, 1 parity or 2 $\leq$ parity cows, respectively, although progesterone concentrations were not different. The presence of fluid-filled luteal cavitied or multiple CL (2 or more) did not affect serum progesterone concentration. A large follicles were observed in 67.4% of heifers or cows and the average diameter was 14.1 mm. Greater luteal tissue area attributed higher pregnancy in heifers, but not in cows, although there were no difference on pregnancy rate according to progesterone concentration in heifers or cows. The pregnancy rate of recipients contained a large follicle at embryo transfer was lower than that of recipients not contained. These results show ultrasonic assessment of ovaries in Holstein recipients is a reliable tool to determine the follicle and CL for recipient selection.
Current developments in IVF and animal cloning have resulted in increasing demand for large quantities of oocytes and ovarian follicles at specific stages of development. These medical and scientific needs may be met by developing an optimal culture system for preantral follicles. In this study, we investigated the growth of porcine preantral follicle cultures in different media and in the presence and absence of serum. Follicles were manually dissected from ovaries obtained from prepubertal gilts at a local slaughterhouse, and cultured for 3 days in M199 or NCSU23 medium supplemented with porcine FSH, transferrin, L-ascorbic acid and insulin. Follicle diameters were measured on day 1 and 3 of culture. In Experiment 1, the effect of supplementing culture medium with fetal calf serum (FCS) on porcine preantral follicle growth was examined. In the group of cultures supplemented with FCS, follicle diameter after 3 days of culture, survival rate and antrum formation rate in the FCS group were significantly higher than those of the control group. In Experiment 2, the effects of culture medium (M199 and NCSU23) on follicle growth were compared. Follicle diameters were increased in the M199 group, compared with those in NCSU23 (p<0.05), but we observed no significant differences in survival and antrum formation rates between cultures grown in the two media. In conclusion, supplementation of the culture medium with serum enhances preantral follicle growth and antrum formation, and M199 is superior to NUSU23 for porcine preantral follicle culture in vitro.
The ultrastructure of oocytes during oogenesis and oocyte degeneration associated with follicle cells in female Sinonovacula constricta(Lamarck, 1818) were investigated by electron microscope observations. Ovarian follicles are surrounded by a matrix of vesicular connective tissue cells(VCT cells). VCT cells contain large quantities of glycogen particles and several lipid droplets in their cytoplasm. It is suggested that VCT cells act as a source of nutrients for vitellogenesis during oogenesis. In early vitellogenic oocytes, several coated vesicles, which appear at the basal region of the oocyte, lead to the formation of membrane-bound vesicles via endocytosis. The uptake of nutritive materials in coated vesicles formed by endocytosis appears through the formation of coated pits on the oolemma during vitellogenesis. During the late stage of oogenesis, yolk precursors(yolk granules), mitochondria and lipid droplets are present in the cytoplasm of late vitellogenic oocytes. In particular, proteinaceous yolk granules containing several different components are intermingles and form immature yolk granules. In the mature oocyte, small immature yolk granules are intermingled and form large mature yolk granules. Vitellogenesis occurs through a process of autosynthesis, involving combined activity of the Golgi complex, mitochondria and rough endoplasmic reticulum in the cytoplasm of vitellogenic oocytes. The process of heterosynthesis is where extraovarian precursors are incorporated into oocytes by endocytosis at the basal region of early vitellogenic oocytes before the formation of the vitelline coat. Follicle cells appear to play an important role in vitellogenesis and oocyte degeneration. The functions of attached follicle cells to the oocyte during oocyte degeneration are phagocytosis and digestion of phagosomes originating from oocyte degeneration. After digestion of phagosomes, it is assumed that the function of follicle cells can permit a transfer of yolk precursors necessary for vitellogenesis and allows for the accumulation of glycogen and lipid during oocyte degeneration, which can be employed by vitellogenic oocytes. Follicle cells of S. constricta may possess a lysosomal system for induction of oocyte breakdown and might resorb phagosomes in the cytoplasm for nutrient accumulation during oocyte degeneration.
As a result of intensive breeding, litter size has considerably increased in pig production over the last three decades. This has resulted in an increase in farrowing complications. Prolonged farrowing will shorten the window for suckling colostrum and reduce the chances for high-quality colostrum intake. Studies also agree that increasing litter sizes concomitantly resulted in decreased piglet birth weight and increased within-litter birth weight variations. Birth weight, however, is one of the critical factors affecting the prognosis of colostrum intake, and piglet growth, welfare, and survival. Litters of uneven birth weight distribution will suffer and lead to increased piglet mortality before weaning. The proper management is key to handle the situation. Feeding strategies before farrowing, management routines during parturition (e.g., drying and moving piglets to the udder and cross-fostering) and feeding an energy source to piglets after birth may be beneficial management tools with large litters. Insulin-like growth factor 1 (IGF-1)-driven recovery from energy losses during lactation appears critical for supporting follicle development, the viability of oocytes and embryos, and, eventually, litter uniformity. This paper explores certain management routines for neonatal piglets that can lead to the optimization of their colostrum intake and thereby their survival in large litters. In addition, this paper reviews the evidence concerning nutritional factors, particularly lactation feeding that may reduce the loss of sow body reserves, affecting the growth of the next oocyte generation. In conclusion, decreasing birth weight and compromised immunity are subjects warranting investigation in the search for novel management tools. Furthermore, to increase litter uniformity, more focus should be placed on nutritional factors that affect IGF-1-driven follicle development before ovulation.
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