Growth Rate Of Follicle Research Articles

Comparison of Bovine Small Antral Follicle Development in Two- and Three-Dimensional Culture Systems.

To compare the effects of two-(2D, microplate) and three-dimensional (3D, alginate) culture systems on the in vitro growth of small antral follicles in cattle, individual follicles were separately cultured in the two culture systems for 8 days. Half of the culture medium was replaced by fresh medium every 2 days; the former medium was used to assess the amount of follicular hormone secretion using ELISA. Individual follicle morphology, diameter, and survival rate were recorded every alternate day. The results showed that in 4 days, there was no significant difference between the two systems, except that the growth rate of follicles in 2D system was relatively faster. After 4 days, estradiol concentration in 3D system was higher than that in 2D system. However, progesterone concentration was lower than that in the 2D system. The survival rate and oocyte quality of follicles in 2D system were significantly lower than those in 3D system on day 8. The follicle diameter slightly increased (30-60 μm) in the entire process. Taken together, for in vitro culture of follicles within 4 days, the 2D culture system is more suitable. However, when the culture duration is >4 days, the 3D culture system is more suitable.

Progesterone Affects Clinic Oocyte Yields by Coordinating with FSH <i>via</i> the PI3K/AKT and MAPK Pathways

Background : Over recent years, progestin has been incorporated into the novel ovary stimulation regimens as an alternative for GnRH analogs to prevent premature luteinizing hormone surge, and has become widely used in treatment for infertility. However, the influence of progesterone administration in terms of clinical outcomes and the mechanisms involved remains poorly understood. Methods: The clinic outcomes from patients undergoing ovary stimulation for in vitro fertilization were analyzed. The mouse ovary stimulation model and follicle culture system were used to evaluate the effect of progesterone on oocyte yield, follicle development, granulosa cell proliferation, hormone secretion. Phospho-Specific Protein Microarray, western blot and RT-PCR were employed to explored the signal mechanism. Findings: We present the evidence from patients suggesting that progesterone decreased the oocyte yields, that was rescued by the higher dose of human menopausal gonadotropin. Using both in vivo and in vitro mouse models, we demonstrated that the administration of progesterone inhibited the proliferation of granulosa cells and therefore reduced the growth rate of follicles. The action of progesterone was mediated by progesterone receptor membrane component-1 and rescued by FSH. Using a phosphorylation array, we identified the PI3K/AKT and MAPK are pivotal pathways that integrated the action of progesterone into the FSH signaling network via the downregulation of cyclin D2 and c-fos in granulosa cells. Interpretation: Our findings highlight the mechanism by which progesterone coordinates with FSH to regulate follicle growth and subsequently influence oocyte outcomes in clinic. These findings advance our understanding of the alterations of ovary response to gonadotrophins during progestin primed ovarian stimulation and gives rise to the opportunity for manipulating individual oocyte yields by signaling regulation. Funding Statement: This study was financially supported by the Natural Science Foundation of Shanghai (Grant number 19ZR1429300, awarded to HL), research grants from the National Natural Science Foundation of China (Grant number 81871163, awarded to QL; Grant number 81771533, awarded to YK; Grant number 81671520, awarded to QC), and the National Key Project (Grant number 2018YFC1003000). Declaration of Interests: The authors have declared that no conflict of interest exists. Ethics Approval Statement: Approval for human retrospective analysis was obtained from the institutional Ethics Committee of Shanghai Ninth People’s Hospital affiliated to Shanghai JiaoTong University’s School of Medicine. Animal studies were approved by the institutional Ethics Committees of Care and Use of Experimental Animals of Shanghai Jiaotong University.

Differential follicle stimulating hormone glycosylation modulates follicle growth and survival rates

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Intrafollicular lipopolysaccharide injection delays ovulation in cows

The aim of this study was to evaluate effects of intra-follicular (i.f.) treatment with lipopolysaccharide (LPS) on follicular and luteal development in cows. There were 18 non-lactating cows assigned to two groups to address this aim: control group (n = 9), which received an i.f. injection of saline; and LPS group (n = 9), which received an i.f. injection of 1 μg of LPS per mL of follicular fluid. Cows were treated with an intravaginal P4 releasing device (IVD) and estradiol benzoate on D0. On D4 and D5 cows were treated with cloprostenol sodium and on D7 the IVD was removed. At 12 h after IVD removal, cows were administered the i.f. injection of LPS or saline. After administration of these treatments, follicular development was evaluated every 12 h until ovulation. The LPS treatment increased blood flow in pre-ovulatory follicles (P = 0.05). Follicle growth was reduced by LPS injection (P < 0.02) resulting a longer period to the time of ovulation for cows in the LPS than control group (P = 0.03). The percentage of cows having ovulations was less for the LPS than control group (P = 0.03). The diameter of the CL, CL blood flow and P4 concentrations 5 and 12 days after ovulation did not differ between groups (P> 0.05). In conclusion, intra-follicular treatment with LPS resulted in a decreased rate of follicle growth, delayed timing of ovulations and a lesser number of cows having ovulations.

Comparison of estradiol benzoate and cypionate in Girolando cows submitted to a timed artificial insemination

The various studies on the manipulation of the estrous cycle culminated in the development of estrous and ovulation synchronization protocols, to the point of defining an optimum moment for artificial insemination. The induction of ovulation is a primordial part for the determination of the moment of the TAI, so to study which hormones and how they behave allows a better understanding of this physiological process and, to manipulate it efficiently. The aim of the present study was to compare the use of estradiol benzoate (EB) and cypionate (ECP) as ovulation inducer in Girolando cows submitted to Timed Artificial Insemination (TAI). Ovulations of 108 Girolando cows were synchronized and the initial day of treatment, recorded as D0, which was when the animals received an intravaginal device containing 1g of progestogen and an intramuscular application (im) of 2mg of EB. After 8 days (D8), the device was removed and 500μg of cloprostenol (im) was administered to all females. Then, the animals were randomly divided into two treatments: BG Group (n = 52) and CG Group (n = 56). The CG animals received 1mg of ECP (im) as the ovulation inducer at the time of device removal, while BG Group cows received 1mg of EB (im) 24 h later (D9). TAI occurred at D10. After device removal, ultrasound evaluations were performed every 12 h up to ovulation. The following reproductive parameters were evaluated: interval from intravaginal device removal to ovulation - IDO (hours); interval from ovulation to TAI-IOT (hours); diameter of the largest follicle at intravaginal device removal (mm); maximum diameter of the DF (mm) at D10; the dominant follicle growth rate (mm/day); synchronization rate - SR (%); ovulation rate - OR (%) and PR - pregnancy rate (%). Only the ovulation rate presented a statistical difference (p&lt;0.05). In conclusion, despite the ovulation rate difference, both estradiol esters administered were effective and presented similar pregnancy rates in Girolando cows submitted to TAI.

The Effect of Radiation Emitted by Cell Phone on The Gelatinolytic Activity of Matrix Metalloproteinase-2 and -9 of Mouse Pre-Antral Follicles during In Vitro Culture.

Objective The unfavorable effects of electromagnetic radiation (EMR) emitted by the cell phone on reproduction health are controversial. Metalloproteinases play a vital role in ovarian follicle development. This study was designed to investigate the effects of exposure to the cell phone on the gelatinolytic activity of in vitro cultured mouse pre-antral follicle. Materials and Methods In this experimental study, pre-antral follicles were isolated from ovaries of immature mice (n=16) and cultured with or without exposure to the cell phone in talking mode for 60 minutes. The gelatinolytic activity was evaluated through the zymography method, as well as the gene expression of matrix metalloproteinases (MMPs) namely MMP-2 and -9 and tissue inhibitors of metalloproteinases (TIMPs) namely, TIMP-1 and -2 by the real-time polymerase chain reaction (PCR) method. Also, in parallel, the development of pre-antral follicles was assessed. Results The maturation parameters of the cell phone-exposed pre-antral follicles were significantly lower compared with the control group (P<0.05). The gelatinolytic activity was significantly decreased in the cell phone-exposed pre- antral follicles compared with the control group (P<0.05). The relative mRNA expression of the MMP-2 gene was significantly (P<0.05) increased in the cell phone-exposed pre-antral follicles whereas the expression rate of the MMP-9 gene was considerably (P<0.05) reduced when compared with the control group. Conversely, the relative expression of the TIMP-1 was markedly (P<0.05) increased in the cell phone-exposed pre-antral follicles while the expression of the TIMP-2 was (P<0.05) significantly diminished in comparison with the control group. ConclusionExposure to the cell phone alters the growth and maturation rate of murine ovarian follicle through the changing in the expression of the MMP-2 and -9 genes, as well as the gelatinolytic activity.

Monitoring follicular dynamics using ultrasonography in captive brown bears (Ursus arctos) during the breeding season

Artificial insemination (AI) may be a useful tool in the reproductive management of endangered animals, including bears. To establish an AI program for bears, we investigated follicular dynamics using weekly transrectal ultrasonography in six captive brown bears. Along with ultrasonography, we monitored plasma progesterone (P4) and estradiol-17β (E2) concentrations. Furthermore, two bears were administered a gonadotropin releasing hormone (GnRH) agonist to induce ovulation on the first day on which the largest follicle reached more than 10.0 mm in diameter. Brown bears showed two patterns of follicular development in the early and late periods of the breeding season. In the early period (May to mid-June), multiple follicular waves were observed; namely, many follicles developed, and the largest follicles grew to less than 6.0 mm in diameter then regressed. In the late period (mid-June to July), one or two follicles grew to greater than 6.0 mm in diameter and developed as dominant follicles. Moreover, the growth rate of the largest follicle in the late period was faster than that in the early period of the breeding season. One bear with a follicle of 13.1 mm ovulated spontaneously, and one bear ovulated when the follicle was 10.2 mm in diameter after GnRH agonist treatment. Plasma E2 concentrations increased and showed peaks five to seven days before the largest follicles reached their maximum size. Plasma P4 concentrations increased on the day the corpus luteum could be detected using ultrasonography. This is the first study that showed there are two patterns of follicular development in brown bears. Furthermore, the largest follicle reaching greater than 10.0 mm in diameter could be an indicator of the appearance of ovulatory follicles.

Mechanisms regulating follicular development and selection of the dominant follicle

Reproductive function is an integrated process encompassing both extra-ovarian signals, such as gonadotrophins, and intrafollicular factors, such as locally produced growth factors. Initiation of primordial follicle growth and the early stages of folliculogenesis can occur without gonadotrophins. However, in vivo and in vitro studies indicate that FSH may stimulate the rate of preantral follicle growth and that it can take only 3 months for a primordial follicle to reach the ovulatory stage. Antral follicle development from 2 and 4 mm in diameter in sheep and cattle, respectively, is gonadotrophin dependent. During the oestrous cycle a transient increase in circulating FSH precedes the recruitment of a group of follicles. Recruited follicles are characterized by induction of expression of mRNAs encoding a range of steroidogenic enzymes, gonadotrophin receptors and local regulatory factors. As follicles continue to mature, there is a transfer of dependency from FSH to LH, which may be part of the mechanism involved in selection of follicles for continued growth. The mechanism of selection of the ovulatory follicle seems to be linked to the timing of mRNA expression encoding LHr and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) in granulosa cells. Locally produced growth factors, such as the insulin-like growth factors (IGFs) and members of the transforming growth factor beta (TGFbeta) superfamily (inhibins, activins and bone morphogenetic proteins (BMPs)), work in concert with gonadotrophins throughout the follicular growth continuum. The roles of growth factors in follicular development and survival are dependent on gonadotrophin status and differentiation state, including morphology. In conclusion, it is the integration of extraovarian signals and intrafollicular factors that determine whether a follicle will continue to develop or be diverted into atretic pathways, as is the case for most of the follicles in monovulatory species, such as cattle.

Effect of oral administration of pioglitazone on follicular dynamics in Holstein dairy cows

This study investigated the effects of oral administration of pioglitazone (PGT), a specific and synthetic ligand of peroxisome proliferator-activated receptors gamma (PPARγ), on follicular dynamics and corpus luteum (CL) functionality in dairy cows. Cows exhibiting strong signs of estrus after 2 injections of PGF2α (given14 d apart) at d 30 postpartum (n = 28) were allotted to four groups (n = 7 cows/treatment) and orally received 6 mg PGT/kg body weight/day according to the following protocol: no PGT (control); PGT for 14 d from 7 d before expected estrus (10 d after 1st injection of PGF2α) to 7 d after observed estrus (PGT14); PGT for 21 d after observed estrus (PGT21); and PGT for 28 d, 7 d before expected estrus to 21 d after observed estrus (PGT28). During the first follicular wave, number of follicles (total and small) increased in PGT14 and PGT28 cows compared to the control group (P < 0.05). During the ovulatory wave, number of total and small follicles increased in PGT28 (P < 0.05) and PGT21 (P < 0.10) compared with PGT14 and control cows. Size of the largest follicle at first wave was greater in PGT28 (P < 0.05), PGT14 (P < 0.05) and PGT21 (P < 0.10) compared to the control cows. Maximal size of the ovulatory follicle was greater in PGT28 (P < 0.05) and PGT21 (P < 0.10) groups compared to the control group. Growth rate of the largest follicle at first wave was higher (P < 0.05) in PGT-treated cows, while growth rate of the ovulatory wave was higher in PGT28 and PGT21 groups, leading to shorter days from luteolysis to ovulation. Pioglitazone administration did not affect CL size, but increased progesterone (P4) concentration. The PGT14 and PGT28 cows had higher maximal plasma P4 concentration and shorter intervals to reach maximal plasma P4 compared to the control group. In conclusion, oral administration of PGT had some positive effects on follicular development and circulating P4 levels which may be conducive to better reproductive performance.

Evaluación ultrasonográfica en ganado Brown Swiss sometido a un protocolo de sincronización de celo en el altiplano peruano

Se evaluó la dinámica folicular mediante ultrasonografía en animales Brown Swiss sometidos a un protocolo de sincronización de celo con progestágenos en el altiplano peruano. Se utilizaron 27 hembras Brown Swiss distribuidas en tres grupos de nueve animales, según su estado fisiológico: vacas en producción (&lt;120 y &gt;260 días de lactancia) y vaquillas. Para la sincronización, en el día 0 se aplicó el dispositivo intravaginal a base de progesterona y se inyectó 2 mg benzoato de estradiol, en el día 8 se extrajo el dispositivo y se aplicó 0.15 mg cloprostenol y 400 UI eCG, y en el día 10 se aplicó 1.05 mg GnRH. La inseminación artificial a tiempo fijo se hizo 12 h después de la aplicación del GnRH. La evaluación ecográfica se hizo los días 8 y 10 para determinar la tasa de crecimiento y diámetro máximo del folículo dominante en los tres grupos. La tasa de crecimiento folicular fue de 2.3 ± 1.5, 2.4 ± 1.8 y 2.3 ± 1.8 mm/día para los grupos de vacas en producción (&lt;120 y &gt;260 días de lactancia) y vaquillas, sin diferencia significativa entre grupos. El diámetro del folículo dominante fue de 14.5 ± 2.1 mm en vacas con &lt;120 días de lactancia, 16.6 ± 3.1 mm en vacas con &gt;260 días de lactancia y 12.1 ± 2.0 mm en vaquillas, siendo significativamente menor en vaquillas (p&lt;0.05). La tasa de preñez fue de 44.4, 33.3 y 66.7% para vacas en &lt;120 días y &gt;260 días de lactancia y vaquillas, respectivamente, sin diferencia significativa entre grupos.

118 A retrospective analysis of follicular dynamics, LH, estradiol-17β, and progesterone in prostaglandin F2α-induced estrus of Beetal goats

The objective of the study was to reveal the factors associated with variable interval to ovulation following a single dose of prostaglandin F2α (PGF2) in Beetal goats. In experiment 1, the extent of ovulation time was demonstrated after synchronizing Beetal goats (n=38) by using a single injection of PGF2 based on corpus luteum detection by B-mode transrectal ultrasonography. Ovulations were detected every 6h from PGF2 until ovulation. To explore the changes in follicular dynamics, endocrine profiles and behavioural events due to the variation in the interval to ovulation, another set of goats (n=13; experiment 2) were synchronized by a single dose of PGF2 given at random days of the luteal phase. Follicular dynamics were monitored every 12h from PGF2 until ovulation and oestrus was detected by apronized bucks every 6h from PGF2 treatment. Blood sampling (3mL) for oestradiol-17β and progesterone was done every 12h from PGF2 until ovulation and every 2h for LH assay by radioimmunoassay. Data were analysed by Student’s t-test, general linear models, and Pearson coefficient correlation (SPSS, version 20.0, SPSS Inc., Chicago, IL, USA). Results revealed that ovulation time varied from 60 to 96h after PGF2, and 57% of ovulations occurred by 72h after PGF2 (experiment 1). Based on this observation, goats (n=13) in second experiment were retrospectively divided into early and late ovulators (i.e. 72 and 84h following PGF2, respectively). Onset of oestrus, peak oestradiol-17β concentration, and LH surge after PGF2 were first observed in early than late ovulators (P&amp;lt;0.05). The goats having greater follicle diameter and smaller corpus luteum diameter at the time of PGF2 ovulated earlier than late ovulators (5.4±0.2v. 4.3±0.2mm and 10±0.6v. 11.8±0.3mm, respectively; P&amp;lt;0.05). Likewise, plasma progesterone concentration tended to be lower (P=0.087) in early than late ovulators. However, interval from onset of oestrus to peak LH and ovulation, duration of standing oestrus, interval from LH peak to ovulation, diameter and growth rate of preovulatory follicles, and number of ovulations were similar (P&amp;gt;0.05) between early and late ovulators. In conclusion, the size of dominant follicle and corpus luteum at the time of PGF2 determines the interval to ovulation following a single dose of PGF2 during the luteal phase.

122 Effect of gonadotropin-releasing hormone plus 7-day progesterone (CIDR) with or without eCG on follicular dynamics, estrus response, and pregnancy rate in anestrous Beetal goats during nonbreeding season under subtropical conditions

Oestrus induction is a challenging task during the nonbreeding season in goats. Therefore, our objective was to evaluate the efficacy of gonadotropin-releasing hormone (GnRH) plus a 7-day CIDR-based protocol with or without eCG for the induction of oestrus and fertility in acyclic Beetal goats during the nonbreeding season under subtropical conditions. Based on the absence of corpus luteum via ultrasonography, goats (n=56; mean weight=32.8±0.5 kg; body condition scre=2.5±0.1) were inserted with a CIDR (300mg progesterone) along with GnRH (Day 0), and randomly treated either with eCG (400 IU; n=35) or without eCG (control; n=21) on Day 7. The CIDR was removed on Day 7 and all does were administered single dose of prostaglandin F2α (75µg cloprostenol; Fatro, Italy). Following CIDR insertion, ovarian follicular dynamics was monitored daily through a 7.5-MHz transrectal transducer (Honda HS 1500, Japan) until ovulation in a subset of eCG (n=11) and control goats (n=10). Oestrus was detected at every 12h after CIDR removal with aproned bucks and does were bred naturally. Pregnancy was detected 25 days after breeding by transrectal ultrasound. Data were analysed by t-test, ANOVA, and Chi-squared test using SPSS (version 20.0, SPSS Inc., Chicago, IL, USA). Oestrus occurred more often following CIDR removal in eCG v. control goats (94v. 57%; P&amp;lt;0.05). Similarly, onset of oestrus was earlier (36.0±2.1v. 54.2±4.7 h; P&amp;lt;0.05) and the duration of oestrus was longer in eCG v. control goats (i.e. 30.2±1.3v. 22.1±2.3 h; P&amp;lt;0.05). However, diameter of preovulatory follicles and interval to ovulation after CIDR removal were similar between eCG and control goats (P&amp;gt;0.05). In contrast, growth rate of preovulatory follicle after CIDR removal until ovulation in eCG goats was greater, and ovulation rate was higher than control goats (0.7±0.1v. 0.3±0.1 mm/day; 2.2±0.2v. 1.4±0.2, respectively; P&amp;lt;0.05). In eCG goats, the dominant follicle grew rapidly between Days 8 to 9 (P&amp;lt;0.05), whereas within control goats the dominant follicle grew at consistent rate (P&amp;gt;0.05). After CIDR removal, percentage of ovulating goats in eCG was higher as compared to the control group (100v. 44%; P&amp;lt;0.05). Within control goats, size of the dominant follicle in ovulatory goats was greater at the time of CIDR removal than anovulatory goats (5.7±0.3v. 5.0±0.15 mm; P&amp;lt;0.05), whereas wave emergence tended to be earlier after CIDR insertion in anovulatory goats (4.2±1v. 2.6±0.4 days; P=0.09). Pregnancy rate 25 days postbreeding did not differ between eCG and control goats [i.e. 49% (16/33) v. 25% (3/12); odds ratio 2.9/1; P&amp;gt;0.05]. In conclusion, the eCG protocol resulted in higher oestrus response, ovulation rate, and 3 times higher pregnancy rate in anestrous goats during the nonbreeding season.

121 Ovulatory response to gonadotropin-releasing hormone relative to day and diameter of dominant follicle during the first follicular wave in Beetal goats

In Beetal goats under subtropical conditions, ovulatory response of first-wave dominant follicle based on diameter and day of oestrous cycle is unknown to date. The objective of this study was to evaluate the ovulatory response to gonadotropin-releasing hormone (GnRH) relative to day and diameter of dominant follicle during the first follicular wave in Beetal goats. Fifty goats were synchronized using a single dose of prostaglandin F2α (75 µg; D+ cloprostenol, Fatro, Italy), and scanned every 6 hours for ovulation using a 7.5-MHz trans-rectal transducer (Honda HS: 1500, Japan). Following ovulation (Day 0), goats (n=40; 10/day) were given GnRH (25µg lecirelin, Fatro, Italy) on Days 2, 4, 6, and 8 of oestrous cycle. Follicular dynamics was monitored daily from D 0 until GnRH, and every 4 hours after GnRH until ovulation. Ovulation frequency relative to day and dominant follicular diameter was analysed by binary logistic regression, whereas diameter of dominant follicle at time of GnRH, ovulation time, time of wave emergence and growth rate of dominant follicle following GnRH were analysed by one-way ANOVA and independent sample t-test (SPSS 20, IBM Corp., Armonk, NY, USA). Eighty percent of the goats ovulated within 74.7±1.3h after prostaglandin F2α. Overall, 37% (14/38) of the goats ovulated following GnRH within 12.0±1.5h. Ovulations (%) following GnRH were higher (P=0.03) on Day 4 than 2 (60 v. 10%; i.e. 14 times higher than Day 2) and decreased gradually from Day 4 to 8 (P&amp;gt;0.05). Similarly, ovulations following GnRH were higher (P=0.00) in first-wave dominant follicles having diameter 6.1-7.0mm than those having less than 6.1mm (88v. 23%; i.e. 24 times higher than &amp;lt;6.1mm). Ovulations (%) decreased as the diameter of dominant follicle increased from 6.1-7.0mm (P&amp;gt;0.05). Diameters of ovulatory follicles at time of GnRH between Day 2, 4, 6, and 8 were not different (P&amp;gt;0.05). Regardless of day of GnRH treatment, diameter of ovulatory v. anovulatory follicles differed (6.44±0.03v. 5.1±0.4 mm; P=0.00). Overall, growth rate of dominant follicles following GnRH was 0.2±0.11mm and did not differ between ovulatory and anovulatory goats (0.29±0.13v. 0.13±0.15 mm; P&amp;gt;0.05). Interval to the emergence of a new wave following GnRH was prolonged for Day 4 compared to Day 2, 6, and 8 (i.e. 22v. 8, 11, and 12 h; respectively, P&amp;lt;0.05). In conclusion, GnRH treatment at Day 4 resulted in maximum ovulations as the first-wave dominant follicle reached a size of 6.1 to 7.0 mm.

Follicle blood flow and FSH concentration associated with variations in characteristics of follicle selection in heifers

Selection of the dominant follicle during a follicular wave is manifested by diameter deviation. At deviation (day 0), growth rate continues for the future dominant follicle (F1) and begins to decrease for the largest subordinate follicle (F2). The percentage of color-Doppler blood-flow signals in the wall of F1 and F2 and the temporality between FSH concentration and F1 and F2 diameter were determined daily in waves 1 and 2 in 24 Holstein heifers. Diameter and blood flow were compared among classes of deviation: (1) conventional (F2 ≥ 7.0 mm on day 0), (2) F2-undersized (F2 < 7.0 mm on day 0), and (3) F1,F2-switched (F2 larger than F1 on days −1 or 0). A class-by-day interaction for diameter of F2 (P < 0.004) and for blood-flow percentage of F2 (P < 0.02) represented greatest values on days −1 or 0 in the switched class and greater values in the conventional than undersized class. Changes were similar between diameter and blood flow in F1 and F2 before deviation. Blood flow in F2 decreased sooner than diameter after deviation indicating that a decrease in vascular perfusion preceded a decrease in diameter. Relationships between F1 and FSH in conventional deviation were similar between waves 1 and 2 for (1) growth rate of F1 on days −1 to 0, (2) interval from emergence of F1 at 4 mm to deviation, and (3) decrease in FSH on days −2 to 0. Relationships between F2 diameter and FSH were dissimilar between classes and between waves 1 and 2 indicating other hormones or factors are also involved in the complex control of F2. For example, the growth rate of F2 was greater (P < 0.05) for conventional than undersized class during wave 1 but similar between classes during wave 2. The FSH surge 2 was similar in profile and prominence between classes but the interval from the FSH peak of surge 2 to deviation was shorter (P < 0.05) in the undersized class (1.5 ± 0.3 d) than in the conventional class (2.3 ± 0.3 d). This was a novel finding and accounted for some of the dissimilarities in growth rate of follicles between classes in wave 2. Results did not support the hypothesis that the extent of blood flow in the wall of future dominant and largest subordinate follicles before deviation is an earlier indicator of follicle destiny than diameter. Results supported the hypothesis that follicle dynamics and FSH concentrations before deviation are temporally associated within conventional and undersized deviation classes but the temporality differs between classes.

In vitro culture of isolated preantral and antral follicles of goats using human recombinant FSH: Concentration-dependent and stage-specific effect

The present study aimed to investigate a concentration-response curve of human recombinant FSH (hrFSH) for in vitro culture of isolated preantral and early antral follicles of goats. Isolated follicles were cultured for 18 days using the following treatments: basic culture medium (control); or control medium supplemented with 10, 50, and 100 mIU/mL of hrFSH. At the end of the culture, cumulus-oocyte complexes were recovered and subjected to in vitro maturation. The following endpoints were evaluated: follicle morphology, growth rate and antrum formation, oocyte viability and meiotic stage, and estradiol production, as well as relative expression of FSH receptor (FSHR), and steroidogenic enzyme (3β-HSD, CYP17, and CYP19A1) genes. In antral follicles, the FSH addition at 50 mIU/mL increased follicular diameter and growth rate, percentage of fully developed oocytes, and oocyte diameter (P < 0.05), and tended to increase the percentage of MII oocytes when compared to the control (P = 0.07). With preantral follicles, FSH addition at 100 mIU/mL increased relative abundance of mRNA for CYP19A1 when compared to the control (P < 0.05). At the same FSH concentrations of 100 and 50 mIU/mL, there was a greater relatively abundance of mRNA for 3β-HSD and CYP17 in preantral than in antral follicles (P < 0.05). For preantral and antral follicle comparisons when the same treatments were imposed, there were greater concentrations of estradiol for antral follicles (P < 0.05). In conclusion, hrFSH enhanced in a concentration-dependent manner the in vitro development of caprine antral follicles; however, there was no positive effect in the culture of preantral follicles.

Influence of Follicular Growth Stimulation on Fertility of Beef Cows Treated with eCG or FSH in FTAI Protocols

Background: The pregnancy rate in fixed-time insemination (FTAI) programs in beef cattle is affected by several factors, including the size of ovulatory follicle (OF). Induced ovulation of small OF (≤11 mm) reduces the chance of pregnancy after FTAI when compared to ovulation of larger follicles (≥12 mm). Hormonal treatments to enhance the follicle growth have been widely used in comercial beef cattle systems, aiming to improve the development of a dominant follicle. The objective of this experiment was to evaluate the effect of treatment with eCG or FSH at the time of progesterone implant removal on the diameter of OF, final follicle growth and pregnancy rate in beef cows.Materials, Methods &amp; Results: Three hundred and seven suckled beef cows with body condition score 2.80 ± 0.24 (1 to 5 scale) and 45 to 70 days postpartum were synchronized to FTAI. On day 0, intravaginal implants containing 1 g of progesterone, were inserted and administered 2 mg of estradiol benzoate (EB), i.m. On day 7, intravaginal implants were removed and 150 µg of D-cloprostenol were administered i.m. At this time, three groups were assigned according to the treatments: eCG (400 UI eCG, i.m.), FSH (25 UI FSH, i.m.) and Control (1 mL Saline, i.m.). On day 8, all cows were injected with 1mg EB i.m. and were inseminated 52 to 56 h after the implant removal. Transrectal ultrasonography exams of ovaries were performed at the time of implant removal and before FTAI to determine the diameter of the largest follicle and the follicular daily growth. Pregnancy exams were conducted by transretal ultrasonography 30 days after FTAI. Statistical analyzes were performed by Chi-square test for pregnancy rate. Ovarian diameters and follicular daily growth were analyzed by ANOVA with means compared by Tukey test considering significant P value &lt; 0.05. The diameter of the largest follicle measured on day 7 (10.07, 9.64 and 10.36 mm) and on day 9 (12.73, 11.53 and 12.57 mm), to eCG, FSH and Control respectively, did not differ within treatments (P &gt; 0.05). Final follicular growth was greater (P &lt; 0.05) on eCG group (1.33 mm/day) compared to Control (1.1 mm/day) and FSH (0.94 mm/day). The pregnancy rates of FTAI were 56.6% (60/106), 52% (51/98) and 50.4% (52/103) to eCG, FSH and Control, respectively, with no statistic difference (P &gt; 0.05).Discussion: Suckled beef cows managed under grazing systems frequently have reduced follicular development and longer postpartum anestrous. In this situation, protocols to FTAI combining progesterone, estradiol and eCG or FSH at the time of progesterone device removal are encouraged to improve final follicular growth, ovulation and to increase conception rate. The hypotesis of similar follicular growth and pregnancy rate in suckled beef cows treated with eCG or FSH was not confirmed in the field trial. eCG treated cows had greater follicular growth, follicular diameter at day 9 and higher pregnancy rate compared to FSH and Control cows. FSH produced similar follicular growth rates to Controls and both were significantly smaller than observed in cows receiving eCG. Also FSH supplementation failed to enhance pregnancy rate compared to Control cows. The longer half-life of eCG and its greater LH-like effects could explain the difference in final follicular stimulation when compared to FSH in FTAI protocols. Thus, administration of a single dose of FSH at progesterone device removal failed to be an alternative to eCG in FTAI synchronization programs.

Ultrasonographic guided follicular dynamics in Gaddi goats during non-breeding season

Goat rearing is an important component of the agrarian society of Himachal Pradesh, India. Gaddi goats constitute about three-fourth of the total goat population of the state and are reared by approximately 35% traditional Gaddi families as migratory flocks on sub-mountainous pastures during winter and high altitude alpine pastures during summers. Out of season breeding is practiced by shepherds which however, needs to be validated with follicular dynamics and endocrine estimations. Adult, healthy, non-pregnant Gaddi does (11), aged 3.74±0.28 years, weighing 24.27±0.70 kg were investigated during conventional non-breeding season of long days (May-June 2016) at University Livestock Farm of CSK Himachal Pradesh Krishi Vishvavidyalya, Palampur for follicular wave characteristics. The follicular wave pattern comprised either three (81.8%, 9) or four (18.2%, 2) waves, the average being 3.18±0.12 during non-breeding seasons with none of these being ovulatory. The data combined for all waves during non-breeding season revealed an average inter wave interval of 5.66±0.28 days, the average growth rate and maximum size of dominant follicles was 0.65±0.03 mm per day and 7.66±0.10 mm, respectively. The dominant follicle (DF) persisted for lesser duration in majority of four wave animals than three wave animals with second wave exhibiting significantly lower (10.0±3.0 vs 13.78±0.57) duration of persistence of DF. Significant difference in the average daily count of small, medium and large follicles were observed in present study. The average daily progesterone concentration during the period of investigation was 0.30±0.04 ng/ml. In conclusion, out of seasonal breeding, could not be supported by repeated ovarian ultrasonography examination in Gaddi goats during long days revealing recurrent follicular waves without ovulation.

Role of progesterone concentrations during early follicular development in beef cattle: II. Ovulatory follicle growth and pregnancy rates

Two experiments were conducted to investigate the role of relatively lesser and greater progesterone (P4) concentrations during early follicular development on ovulatory follicle growth and pregnancy rate in beef cattle. In Experiment 1, time of ovulation was synchronized with the 5 d CO-Synch + CIDR (Controlled Internal Drug Release) program in multiparous cows (n = 241). Six days after the 2nd GnRH injection of the pre-synchronization program (d 0), ablation of follicles ≥ 5 mm in the ovaries was performed and cows were assigned to receive either a previously used CIDR and 2x-25 mg PGF2α doses 8 h apart (LoP4), or a new CIDR (HiP4). On d 5, CIDR were removed from all cows, 2x-25 mg PGF2α were administered, and estrous detection tail paint was applied. Timed artificial insemination (TAI) was performed on d 8. On d 5, P4 concentrations were greater (P < 0.01) in the HiP4 (4.9 ± 0.13 ng/mL) than LoP4 (1.0 ± 0.06 ng/mL) treatment group. Conversely, d 5 estradiol (E2) concentrations and follicular diameter were greater (P < 0.01) in the LoP4 (5.0 ± 0.23 pg/mL and 8.9 ± 0.20 mm) than HiP4 (1.5 ± 0.12 pg/mL and 7.4 ± 0.15 mm) treatment group. Follicular diameter at TAI (12.0 ± 0.12 mm, Table 1) and TAI pregnancy rate did not differ (P > 0.10) between treatment groups. In Experiment 2, a new follicular wave was induced with estradiol benzoate on d -7, and cows (n = 275) were assigned on d 0 to receive 25 mg PGF2α and either have the CIDR replaced with a new CIDR (HiP4) or the used CIDR was left in place (LoP4).Furthermore, all cows received GnRH on d 0. The CIDRs were removed from all cows on d 5 and two doses of -25 mg PGF2α were administered. Estrous detection combined with AI 12 h later (Estrus-AI) was performed for 60 h after CIDR removal with TAI coupled with GnRH administration at 72 h if estrus was not detected. The concentrations of P4 on d 5 were greater (P < 0.01) in the HiP4 (2.8 ± 0.10 ng/ml) than LoP4 (1.7 ± 0.05 ng/mL) treatment group. For cows that were detected in estrus after PGF2α administration, estrous response (83.5%) and interval to estrus (55.0 ± 0.5 h) did not differ between treatment groups. Pregnancy rate (combined Estrus-AI and TAI) that resulted from breeding at the time of the synchronized time of estrus was similar between treatment groups (HiP4: 77.1%; LoP4: 82.3%). In conclusion, differences in P4 concentrations during early follicular development do not effect pregnancy rate in beef cows when the cows are inseminated at the time of a synchronized estrus if the cows have similar intervals of proestrus.

Follicle Deviation and Growth Rate in Mules

Follicle Deviation and Growth Rate in Mules

Spatial Distribution of Preantral Follicles in the Equine Ovary

The aim of this study was to evaluate effects of intra-follicular (i.f.) treatment with lipopolysaccharide (LPS) on follicular and luteal development in cows. There were 18 non-lactating cows assigned to two groups to address this aim: control group (n = 9), which received an i.f. injection of saline; and LPS group (n = 9), which received an i.f. injection of 1 μg of LPS per mL of follicular fluid. Cows were treated with an intravaginal P4 releasing device (IVD) and estradiol benzoate on D0. On D4 and D5 cows were treated with cloprostenol sodium and on D7 the IVD was removed. At 12 h after IVD removal, cows were administered the i.f. injection of LPS or saline. After administration of these treatments, follicular development was evaluated every 12 h until ovulation. The LPS treatment increased blood flow in pre-ovulatory follicles (P = 0.05). Follicle growth was reduced by LPS injection (P < 0.02) resulting a longer period to the time of ovulation for cows in the LPS than control group (P = 0.03). The percentage of cows having ovulations was less for the LPS than control group (P = 0.03). The diameter of the CL, CL blood flow and P4 concentrations 5 and 12 days after ovulation did not differ between groups (P> 0.05). In conclusion, intra-follicular treatment with LPS resulted in a decreased rate of follicle growth, delayed timing of ovulations and a lesser number of cows having ovulations.