Exogenous progesterone enhances ova and embryo quality following superstimulation of the first follicular wave in Nelore (Bos indicus) donors

Fertility of lactating dairy cows is associated with reduced progesterone (P4) concentration compared with nonlactating animals. The objective of the current study was to determine whether P4 during growth of the first follicular wave (FFW) affects embryo quality. Lactating Holstein cows at 33±3 days post partum were allocated to one of three treatments. Cows in the FFW and FFW with P4 (FFWP) treatments started the superstimulation protocol on day 1 of the estrous cycle and second follicular wave (SFW) cows started the superstimulation protocol on estrous cycle day 7. Cows were superstimulated with 400 mg of NIH-FSH-P1 (FSH) given twice daily for 5 days, two prostaglandin F2α (PGF2α) injections given with the ninth and tenth injections of FSH, GNRH given 48 h after the first PGF2α injection, and timed insemination 12 and 24 h after the GNRH injection. Cows in the FFWP treatment received two intravaginal P4 inserts during the superstimulation. Embryos were recovered 6.5 days after artificial insemination and excellent/good and fair embryos were frozen and transferred. Blood was sampled daily from estrous cycle day 0 until insemination from donor cows. During the superstimulation protocol, P4 was (P<0.01) greatest for SFW cows followed by FFWP and FFW cows respectively. The percentage of embryos–oocytes from SFW and FFWP cows classified as excellent/good and fair embryos was (P=0.02) greater than those of FFW cows. Pregnancy per embryo transfer was not (P≥0.73) affected by embryo donor treatment. Reduced embryo quality of cows induced to ovulate the follicles from the first follicular wave is a consequence of reduced P4 during follicle growth.

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273 OVUM PICKUP AT DIFFERENT TIMES OF A SYNCHRONIZED FOLLICULAR WAVE DID NOT AFFECT IVP IN BOS INDICUS, BOS TAURUS, OR BUBALUS BUBALIS

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab273 ◽

2010 ◽

Vol 22 (1) ◽

pp. 293 ◽

Cited By ~ 5

Author(s):

L. U. Gimenes ◽

M. L. Ferraz ◽

A. Araujo ◽

P. Fantinato Neto ◽

M. R. Chiarati ◽

...

Keyword(s):

Mixed Model ◽

Bos Taurus ◽

Bos Indicus ◽

Cumulus Cell ◽

Bubalus Bubalis ◽

Genetic Group ◽

Cleavage Rate ◽

Follicular Wave ◽

Counting Data ◽

Initial Hypothesis

One important factor in the success of ovum pickup (OPU)/IVP in Bos taurus is the follicular status at OPU concerning the dominance period (Hendriksen et al. 2000 Theriogenology 53, 11-20). The hypothesis of the present study is that OPU performed after follicle deviation, when follicles show a mild level of atresia, improves competence for IVP in Nelore (NE), Holstein (HO), and buffaloes (BU). Objectives were to determine effects of OPU done at different times of synchronized follicular wave (1, 3, or 5 d after expected emergence) and of genetic group (NE, HO, and BU) on IVP. A total of 27 heifers (9 of each genetic group) were maintained in contemporary nutritional and environmental conditions during experiment, in a cross-over design, performed in 6 replicates. Recovered oocytes with at least one cumulus cell layer were matured in TCM-199 supplemented with 10% of FCS plus 50 μM of cysteamin and 0.3 mM of cystine, at 38.5°C with 5% CO2 in air for 24 h. IVF was done with 2 × 106 spermatozoa per mL of NE (for bovine oocytes) or BU semen (for BU oocytes), for 20 h at the same incubator conditions of IVM. After IVF, presumptive zygotes were denuded and cultured in SOF under the same previous atmosphere conditions. Medium was changed 3 d after IVF when cleavage rate (CR) was assessed. Blastocyst (BR) and hatching rates (HR) were evaluated 7 and 9 days after IVF, respectively. About 50% of hatched blastocysts were fixed until nuclei counting. Data were analyzed by ANOVA using the Proc Mixed model. No effects of interaction or time of synchronization were observed in any of the variables. Concerning genetic group, NE had better results than HO and BU (mean ± SEM / heifer / replicate), respectively, for visualized follicles (41.0a ± 2.1, 22.1b ± 1.3, 18.8b ± 0.9), total oocytes (37.1a ± 2.5, 15.4b ± 1.2, 14.8b ± 1.0), oocytes at IVM (30.8a ± 2.4, 10.7b ± 1.0, 7.9b ± 0.7), oocytes at IVC (18.7a ± 0.8, 8.0b ± 0.5, 7.5b ± 0.4), cleaved embryos (15.4a ± 0.7, 4.6b ± 0.4, 4.4b ± 0.3),CR(81.8a, 59.1b, 62.3b), blastocysts on Day 7 (5.1a ± 0.6, 1.0b ± 0.2, 0.6b ± 0.1), BR (25.8a, 13.6b, 9.1b), and hatched blastocysts on Day 9 (2.6a ± 0.4, 0.3b ± 0.1, 0.3b ± 0.1). Recovery rate and HR were greater for NE (89.4 and 50.6%, respectively) than for HO (73.3 and 23.2%), but neither differed from BU (82.8 and 31.9%). Also, the percentage of viable was greater for NE (83.0) than for HO (66.9) and BU (53.1). No effects were observed for nuclei counting (NE = 176.6 ± 5.3, HO = 168.9 ± 13.7 and BU = 206.1 ± 23.1). Results demonstrate that Nelore had a better efficiency for IVP than Holstein and buffaloes. OPU performed at different times of synchronized follicular wave did not influence IVP, conversely to the initial hypothesis of this study. FAPESP (06/59550-6, 07/04782-2), Tortuga Cia Zootecnica®, Santa Adele and São Caetano Farms, LMMD, PCAPS, HOVET (Dr. Ubiraem Schalch), VRA, VNP (Prof. Dr. Francisco de Palma Rennó).

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155 CHARACTERIZATION OF LIPID DROPLETS IN BOS INDICUS AND BOS TAURUS EMBRYOS

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab155 ◽

2013 ◽

Vol 25 (1) ◽

pp. 226 ◽

Cited By ~ 1

Author(s):

E. P. López-Damián ◽

T. Fiordelisio ◽

M. A. Lammoglia ◽

M. Alarcón ◽

M. Asprón ◽

...

Keyword(s):

Lipid Droplets ◽

Embryo Quality ◽

Developmental Stages ◽

Bos Taurus ◽

Inner Cell Mass ◽

Cell Mass ◽

Bos Indicus ◽

Nile Red ◽

Blastocyst Stage ◽

Transfer Program

Accurate evaluation of bovine embryos for assessing developmental stage and quality is critical to the success of any embryo transfer program. However, this evaluation process has been reported to be highly subjective in Bos indicus (BI) and can vary as much as 23% compared with that of Bos taurus (BT). These differences in assessment may be related to the quantity of lipid droplets (LD) within the embryo, which has been shown to have a negative effect in cryopreserving embryos. The aim of the present study was to characterize the number and size of LD in different developmental stages of fresh embryos from BI and BT and to compare LD across the three different embryo quality grades (1 = excellent or good, 2 = fair, and 3 = poor). Nonsurgical embryo collection was performed 7 days post-insemination in 10 BI and 10 BT females. Forty-eight embryos were evaluated for stage and grade using stereoscopic microscopy, processed for transmission electron microscopy, and stained with Nile red. Digitalized images were analyzed with ImageJ (National Institutes of Health, Bethesda, MD, USA), contour of lipid droplets were designed, and values of perimeter, area, and fluorescence intensity were assessed. Nonparametric statistical analysis (Mann–Whitney) was utilized. There was no difference in LD number for BT or BI for morulae and blastocyst; however, BI morulae presented larger LD compared with blastocyst stage embryos (286 µm2 v. 223 µm2; P < 0.05). Likewise, BI TF cells had more LD compared with inner cell mass (ICM) cells (48 v. 36; P < 0.05). BT TF cells exhibited larger LD compared with ICM cells (149 µm2 v. 128 µm2; P < 0.05), while BI embryos exhibited a larger area of LD in the ICM compared with the TF (591 µm2 v. 472 µm2; P < 0.05). In all embryos, BI contained more lipid droplets than BT (78 v. 49; P < 0.05). Across all quality grades (good, fair, and poor) there was no difference in the number of LD in BT embryos; however, BI grade-3 embryos presented more LD than grade-1 (36 v. 25). BT embryos LD were larger than BI LD (907 µm2 v. 625 µm2; P < 0.05). Fluorescence images showed higher arbitrary units of fluorescence (auf) for LD in BI. Compared with BT embryos (386 auf v. 280 auf; P < 0.05). These results suggest that BI embryos contain more and smaller LD than BT embryos and the LD described for BI embryo quality grade 1 are larger than those of quality grades 2 and 3, and even though the number of LD in morulae and blastocyst stage embryos are not different LD size is reduced as development occurs. Research funding provided by UNAM-DGAPA-PAPIIT IN200810.

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347 COULD THE DIFFERENTIAL EXPRESSION OF LUTEINIZING HORMONE RECEPTOR ISOFORMS EXPLAIN THE VARIABILITY IN SUPEROVULATORY RESPONSES IN CATTLE?

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab347 ◽

2015 ◽

Vol 27 (1) ◽

pp. 261

Author(s):

S. Wohlres-Viana ◽

E. K. N. Arashiro ◽

J. G. V. Grazia ◽

L. S. A. Camargo ◽

M. A. Machado ◽

...

Keyword(s):

Rna Extraction ◽

Bos Indicus ◽

Luteinizing Hormone Receptor ◽

Poor Responders ◽

Embryo Production ◽

Lh Receptor ◽

Follicular Wave ◽

Follicular Aspiration ◽

Receptor Isoforms

Embryo production in vivo is highly variable among donors. The Gir breed (Bos indicus) is well known to show a low embryo production after superovulation (2.5 to 3.5 viable embryos per flush), and a high variance in superovulatory responses, which makes this breed an interesting model to study this trait. The aim of this study was to evaluate the expression pattern of LHR isoforms in Gir heifers previously characterised as good (10.3 ± 1.2 embryos/flush, N = 5) or poor (1.1 ± 0.3 embryos/flush, N = 5) responders to superovulation protocols. In both groups, an adapted ultrasound-guided follicular aspiration system (Arashiro et al. 2012 Reprod. Fertil. Dev. 24, 175) was used to collect granulosa cells (GC) from 8-mm follicles growing in either a synchronized but not stimulated follicular wave (FW) or in the fourth day of superovulation (SOV), induced with 200 UI of FSHp (Pluset, Serono). The recovered follicular fluid was centrifuged and the cells were washed with NaCl 0.9% saline and kept in RNA Later (Ambion, Austin, TX, USA). Total RNA extraction was performed using the commercial RNeasy Micro Kit (Qiagen, Valencia, CA, USA). The RNA samples were quantified and reverse transcribed using the commercial Superscript III kit (Invitrogen, Carlsbad, CA, USA). Complementary DNA samples were amplified through real-time PCR, using a LH receptor primer – not selective for LHR isoforms (total LHR) – and 4 sets of isoform selective primers (S1, S10, S10+11, and S11). All samples were previously tested for theca cell contamination through detection of CYP17A1 gene, and those showing contamination were excluded. The β-actin gene was used as endogenous control. Analyses were performed using the REST software and the expression values are shown as mean ± s.e.m. For comparisons between good and poor responders, the first was set as 1.00. For comparisons between FW and SOV, FW was set as 1.00. In the good responder group, there was no difference (P > 0.05) in total LHR expression among GC samples from FW and SOV. However, the S10+11 isoform was down-regulated (0.4 ± 0.1; P < 0.01) after SOV. In the poor responders group, total LHR expression was down-regulated (0.2 ± 0.1; P < 0.01) after SOV, but there was no difference in the expression of isoforms (P > 0.05). Contrasting the response groups (good and poor), total LHR (15.1 ± 7.6; P < 0.001), and the isoforms S10 (5.7 ± 2.7; P < 0.01), S10+11 (1.9 ± 0.6; P < 0.01), and S11 (5.1 ± 2.5; P < 0.01) were up-regulated in FW of poor responders, but there was no difference (P > 0.05) in any LHR form during SOV. We concluded that 1) LHR expression is different between heifers characterised as good or poor responders to superovulation; 2) superovulation modulates the LHR expression and reduces the original differences observed in unstimulated cycles; 3) diminished expression of total LHR, but not in the isoforms, in poor responders heifers could suggest a reduction in the expression of full-length LHR, with possible consequences to ovulatory capability after superovulation.Financial support was provided by CNPq Project 477701 and Fapemig PPM 0067/11.

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Effect of superstimulatory treatments on the expression of genes related to ovulatory capacity, oocyte competence and embryo development in cattle

Reproduction Fertility and Development ◽

10.1071/rd12271 ◽

2013 ◽

Vol 25 (1) ◽

pp. 17 ◽

Cited By ~ 16

Author(s):

Ciro M. Barros ◽

Rafael A. Satrapa ◽

Anthony C. S. Castilho ◽

Patrícia K. Fontes ◽

Eduardo M. Razza ◽

...

Keyword(s):

Mrna Expression ◽

Granulosa Cells ◽

Embryo Quality ◽

Bos Taurus ◽

Bos Indicus ◽

Treatment Protocol ◽

Cumulus Cells ◽

Fixed Time ◽

Treatment Protocols ◽

Oocyte Competence

Multiple ovulation (superovulation) and embryo transfer has been used extensively in cattle. In the past decade, superstimulatory treatment protocols that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-time AI (FTAI), have been developed for zebu (Bos indicus) and European (Bos taurus) breeds of cattle. There is evidence that additional stimulus with LH (through the administration of exogenous LH or equine chorionic gonadotrophin (eCG)) on the last day of the superstimulatory treatment protocol, called the ‘P-36 protocol’ for FTAI, can increase embryo yield compared with conventional protocols that are based on the detection of oestrus. However, inconsistent results with the use of hormones that stimulate LH receptors (LHR) have prompted further studies on the roles of LH and its receptors in ovulatory capacity (acquisition of LHR in granulosa cells), oocyte competence and embryo quality in superstimulated cattle. Recent experiments have shown that superstimulation with FSH increases mRNA expression of LHR and angiotensin AT2 receptors in granulosa cells of follicles >8 mm in diameter. In addition, FSH decreases mRNA expression of growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) in oocytes, but increases the expression of both in cumulus cells, without diminishing the capacity of cumulus–oocyte complexes to generate blastocysts. Although these results indicate that superstimulation with FSH is not detrimental to oocyte competence, supplementary studies are warranted to investigate the effects of superstimulation on embryo quality and viability. In addition, experiments comparing the cellular and/or molecular effects of adding eCG to the P-36 treatment protocol are being conducted to elucidate the effects of superstimulatory protocols on the yield of viable embryos.

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Effect of combined exogenous progesterone with luteotrophic support via equine chorionic gonadotrophin (eCG) on corpus luteum development, circulating progesterone concentrations and embryo development in cattle

Reproduction Fertility and Development ◽

10.1071/rd14019 ◽

2016 ◽

Vol 28 (3) ◽

pp. 269 ◽

Cited By ~ 11

Author(s):

L. O'Hara ◽

N. Forde ◽

P. Duffy ◽

F. Randi ◽

A. K. Kelly ◽

...

Keyword(s):

Corpus Luteum ◽

Embryo Quality ◽

Single Injection ◽

Chorionic Gonadotrophin ◽

Serum Progesterone ◽

Conceptus Development ◽

Luteal Tissue ◽

Exogenous Progesterone ◽

Intravaginal Device

The aim was to examine the effect of a single intramuscular (i.m.) injection of equine chorionic gonadotrophin (eCG) on Day 3 after oestrus on corpus luteum (CL) development, circulating progesterone and conceptus development in cross-bred beef heifers. In Experiment 1, heifers received: (1) saline, or a single i.m. injection of eCG on Day 3 at (2) 250 IU (3) 500 IU (4) 750 IU or (5) 1000 IU. Administration of eCG resulted in increased luteal tissue area and progesterone and oestradiol concentrations compared with controls. In Experiment 2, heifers received (1) a progesterone-releasing intravaginal device (PRID Delta) from Day 3 to 5 or (2) a PRID Delta from Day 3 to 5 plus a single injection of 750 IU eCG on Day 3. In vitro-produced blastocysts (n = 10 per recipient) were transferred on Day 7 and heifers were slaughtered on Day 14 to assess conceptus development. Administration of eCG reduced the number of short cycles (6.3% vs 31.3%) and increased mean luteal tissue weight (P = 0.02). Insertion of a PRID Delta on Day 3 resulted in an elevation (P < 0.05) in serum progesterone until removal on Day 5. Administration of eCG at the time of PRID Delta insertion resulted in higher progesterone levels (P < 0.05) from Day 10 onwards. Conceptus dimensions were not affected. In conclusion, a single injection of eCG on Day 3 increased CL size and progesterone concentrations and, when given in conjunction with a progesterone-releasing device, appeared to reduce the number of short cycles, presumably due to its luteotrophic nature. The implications of the elevated oestradiol concentrations for embryo quality require further study.

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Comparison of three doses of estradiol benzoate for synchronization of follicular wave emergence in suckled Bos indicus beef cows

Animal Reproduction ◽

10.1590/1984-3143-ar2021-0016 ◽

2021 ◽

Vol 18 (3) ◽

Author(s):

Amanda Guimarães Silva ◽

Leonardo Marin Ferreira Pinto ◽

Nadark de Amorim Silva ◽

Ana Clara Degan Mattos ◽

Pablo Henrique Ambrósio ◽

...

Keyword(s):

Bos Indicus ◽

Beef Cows ◽

Estradiol Benzoate ◽

Follicular Wave

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Follicular dynamic and repeatability of follicular wave development in Peranakan Ongole (PO) cattle

Jurnal Ilmu Ternak dan Veteriner ◽

10.14334/jitv.v21i1.1349 ◽

2016 ◽

Vol 21 (1) ◽

pp. 26 ◽

Cited By ~ 1

Author(s):

Muhammad Imron ◽

Iman Supriatna ◽

. Amrozi ◽

Mohamad Agus Setiadi

Keyword(s):

Bos Taurus ◽

Research Result ◽

Bos Indicus ◽

Follicular Development ◽

Wave Pattern ◽

Follicular Wave ◽

Follicle Diameter ◽

Wave Patterns ◽

The Difference ◽

Follicular Dynamics

<p class="abstrak2">Superovulation treatment on PO cattle (Bos indicus) was less responsive compared to Bos taurus breed. It might due to the difference of their follicular dynamic. This study was conducted to investigate the follicular dynamics and its repeatability in PO cattle. Follicular dynamics observations conducted on 9 cows trough ultrasound scanning every day. Observations of wave patterns repeatability were performed in 6 cows which its wave pattern already known on the next consecutive IOI. Research result indicated that PO cattle had 3 (66%) and 4-waves (34%) pattern. The first wave of 3 and 4-waves pattern emerged on day -0.4+0.9 and 1.4+1.1 respectively. The second wave of 3 and 4-wave pattern emerged on day 9.8+1.5 and 7.4+1.9 respectively. The pattern of 3 waves has a longer follicle dominant duration (11.6+1.5 day) in the first wave of estrous cycle, compared with 4 waves pattern (10+2.92 and 7+1.00 day respectively). The growth rate of dominant follicle was not different significantly between the 3 and 4-waves pattern (0.87+0.23 and 0.94+0.25 mm/day respectively). Similarly, ovulatory follicle diameter between 3 and 4-waves pattern was also not different significantly (12.24+12.34 and 12.30+12.23 mm respectively). Observation of wave patterns repeatability in 6 PO cows indicated that PO cattle had high repeatability in follicular wave pattern (0.88) and the number of growing follicle was 0.91. This study resulted data for dynamic of follicular development, wave pattern, its repeatability which be expected to design the protocol of superovulation treatment or other reproduction technologies based on follicular dynamic to improve its result in PO cattle.</p><p> </p>

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PSX-6 The appropriate phase of follicle size and progesterone concentration could control ovulation of ruminant animal

Journal of Animal Science ◽

10.1093/jas/skz258.924 ◽

2019 ◽

Vol 97 (Supplement_3) ◽

pp. 469-469

Author(s):

Junyou LI ◽

Chunxiang Piao ◽

Min Ma ◽

Masayoshi Kuwahara

Keyword(s):

Follicular Development ◽

Progesterone Concentration ◽

Estrous Cycles ◽

Follicular Waves ◽

Follicular Wave ◽

Ruminant Animal ◽

Prostaglandin F2a ◽

Normal Body Weight ◽

Follicle Size ◽

Exogenous Progesterone

Abstract Ovarian follicular development was monitored daily by transrectal ultrasonographic examination in 20 female goats for 2 successive estrous cycles (40 cycles). These animals exhibited ovulatory cycles with a mean length of 20.8 ± 1.1 days. The mean number of follicles ovulated was 2.3 ± 0.9 and their diameter before ovulation was 6.9 ± 1.1 mm. Among 40 estrous cycles examined, 9 (23 %) displayed 1 latent follicular wave and the remaining 31 (77 %) cycles displayed 2 latent follicular waves. Three and 4 days after the second spontaneous ovulation (day 0 = ovulation), when diameter of dominant follicles of the first latent wave exceeded 5.5 mm, 20 goats were treated with a luteolytic dose of prostaglandin F2a (PGF2a) and randomly assigned to one of three groups. This treatment immediately decreased plasma levels of progesterone and increased those of estradiol, resulting in ovulation 2 or 3 days later. One group of goats (n = 5) treated with PGF2a at the first latent follicular wave and was mated with fertile male goat, resulting in 4 pregnancies. For the second group of goats (n = 5), 4 successive short estrous cycles were induced. A total of 35 short ovulatory cycles were induced at a mean interval of 6.5 ± 1.2 days with a range of 4–9 days. Five of goats were allowed to mate following 4 consecutive short ovulatory cycles; normal oestrous behavior was observed at each estrus. The goats were successfully mated, became pregnant and delivered kids of normal body weight and length. The remaining goats (n = 6), in addition to PGF2a treatment, received exogenous progesterone supplement which allowed latent follicular development to occur, but prevented ovulation until the effect of progesterone supplement subsided. Thus, it was concluded that appropriate phase of follicle size and progesterone concentration could control ovulation of ruminant animal.

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