scholarly journals Relationship of vascular perfusion of the wall of the preovulatory follicle to in vitro fertilisation and embryo development in heifers

Reproduction ◽  
2009 ◽  
Vol 137 (4) ◽  
pp. 689-697 ◽  
Author(s):  
M A R Siddiqui ◽  
E L Gastal ◽  
M O Gastal ◽  
M Almamun ◽  
M A Beg ◽  
...  
Keyword(s):  
Embryo Development ◽  
Follicular Fluid ◽  
Polar Body ◽  
Prostaglandin F2α ◽  
In Vitro Fertilisation ◽  
Preovulatory Follicle ◽  
Cleavage Rate ◽  
Vascular Perfusion ◽  
Cell Versus

The effect of the extent of vascular perfusion of the wall of the preovulatory follicle on in vitro cleavage rate of the recovered oocyte and embryo development to >8 cells was studied in 52 heifers. Heifers received a luteolytic dose of prostaglandin F2α (PGF2α) when the largest follicle was ≥11 mm. An ovulation-inducing injection of GnRH was given 36 h later (hour 0), and collection of follicular fluid and the oocyte was done at hour 26. Vascular perfusion of the follicular wall was assessed by colour Doppler ultrasonography at hours 0 and 26. Each of the recovered oocytes (41/52; 79%) was mature (extruded polar body). Cleavage and embryo development were assessed at 48 h and 120 h respectively, after in vitro fertilisation (IVF). The percentage of cleaved oocytes and >8 cell embryos was 80% (31/39) and 55% (17/31) respectively. Vascular perfusion of the follicular wall was greater (lower pulsatility index; P<0.001) for follicles that produced cleaved versus non-cleaved oocytes and greater (P<0.04) for follicles that produced >8 cell versus ≤8 cell embryos. Percentage of follicular wall with Doppler signals of blood flow was greater (P<0.001) for >8 cell versus ≤8 cell embryos. Follicular-fluid concentration of free IGF1 was lower for cleaved oocytes (P<0.001) and >8 cell embryos (P<0.05), and oestradiol was lower (P<0.05) for >8 cell embryos. Results supported the hypothesis that greater vascular perfusion of the wall of the preovulatory follicle was positively associated with IVF and embryo development.

Follicular fluid supplementation during in vitro maturation promotes sperm penetration in bovine oocytes by enhancing cumulus expansion and increasing mitochondrial activity in oocytes

2012 ◽  
Vol 24 (5) ◽  
pp. 743 ◽  
Author(s):  
Tamás Somfai ◽  
Yasushi Inaba ◽  
Shinya Watanabe ◽  
Masaya Geshi ◽  
Takashi Nagai
Keyword(s):  
Embryo Development ◽  
Follicular Fluid ◽  
Calf Serum ◽  
In Vitro Fertilisation ◽  
Mitochondrial Activity ◽  
Atp Content ◽  
Promoting Effect ◽  
Cumulus Expansion ◽  
Sperm Penetration

The aim of this study was to examine the effects of bovine follicular fluid (bFF) on mitochondrial activity in in vitro-matured (IVM) oocytes and to assess its importance for fertilisation and embryo development. Bovine follicular oocytes were subjected to IVM in medium supplemented either with polyvinylpyrrolidone, bovine serum albumin, calf serum or bFF. Nuclear maturation, cumulus expansion, mitochondrial distribution and ATP content in oocytes were compared between groups along with subsequent in vitro fertilisation (IVF) and embryo development. Compared with other supplements, bFF generated significantly enhanced re-distribution of active mitochondria in oocytes and this effect was associated with elevated intracellular ATP content. Furthermore, bFF significantly improved cumulus expansion, which was associated with improved fertilisation rates when cumulus-enclosed oocytes were subjected to IVF; however, its promoting effect was neutralised when denuded oocytes were inseminated. Elevating ATP content in oocytes by bFF did not affect maturation or embryo development but promoted fertilisation when mitochondrial electron transport was blocked in oocytes before IVF by Rotenone. In conclusion, supplementation of IVM medium with bFF promotes sperm penetration both by the improvement of cumulus expansion and by enhancing ATP levels in oocytes, which maintains their ability to be fertilised after mitochondrial stress.

88 EFFECTS OF REMOVAL OF CUMULUS CELLS AND pb1 PRESENCE OF IN VITRO-MATURED BUFFALO OOCYTES PRIOR TO IVF ON CLEAVAGE RATE AND SUBSEQUENT EMBRYO DEVELOPMENT

2014 ◽  
Vol 26 (1) ◽  
pp. 158
Author(s):  
J.-H. Shang ◽  
H.-Y. Zheng ◽  
C.-Y. Yang ◽  
F.-X. Huang ◽  
B.-Z. Yang ◽  
...  
Keyword(s):  
Embryo Development ◽  
Natural Science ◽  
Polar Body ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Lower Efficiency ◽  
Significant Difference

The efficiency of oocyte maturation and embryo production in vitro in buffalo is relatively poor when compared with that in cattle. The percentage of oocytes selected by pb1 (the 1st polar body) presence for somatic cell nuclear transfer (SCNT) ranged from 50 to 70% in our laboratory, which meant that 30 to 50% oocytes have been abandoned. The present study was designed to identify the effect of cumulus cells removal and pb1 presence or absence before the IVF of matured buffalo oocytes on cleavage rate and subsequent embryo development and to try to reuse those oocytes without pb1 for embryo in vitro production. In vitro-matured oocytes enclosed with cumulus cells were randomly selected and denuded mechanically, then the denuded oocytes (DO) were divided into 3 groups by non-selection (pb1 ± ), selection of pb1 presence (pb1+) and absence (pb1–). Intact cumulus–oocyte complexes (COC, control) and pb1 ± , pb1+, and pb1– DO (treatments) were inseminated with motile buffalo sperm in Tyrode's medium for 24 h. The presumed zygotes were washed 3 times and transferred into 50-μL droplets of IVC medium (TCM 199 + 10% fetal bovine serum) and co-cultured with buffalo cumulus cells monolayer for more than 10 days to evaluate the developmental ability of embryos. Cleavage rate (CR) and blastocyst rate (BR) were assessed at 48 h and 240 h after fertilization (0 h). The results indicated that CR and BR for COC (61.69 ± 9.22% and 34.07 ± 7.61%) and pb1+ (66.59 ± 15.50% and 35.96 ± 10.87%) were significantly higher (P < 0.01) than those for pb1 ±  (49.11 ± 6.83% and 21.88 ± 8.17%) and pb1– (35.09 ± 9.17% and 13.16 ± 5.38%). In addition, there was a significant difference (P < 0.05) in the CR and BR between pb1 ±  and pb1– but no difference (P > 0.05) between COCs and pb1+ DO. These data show that removal of cumulus cells before IVF significantly reduces the overall developmental competence to cleavage and blastocyst stage and this negative effects mainly caused by the immature oocytes (indicated by the absence of pb1), but there was no effect on mature oocytes (presence of pb1). However, the oocytes without pb1 can still be used for in vitro embryo production even with lower efficiency when compared with intact COC. This research was supported by grants from the National Natural Science Foundation of China (31160456), the Natural Science Foundation of Guangxi, China (2011GXSFB018045, 2013GXNSFAA019075).

188 HAPLOID ACTIVATION OF BOVINE OOCYTES WITH IONOMYCIN AND SINGLE OR COMBINED ACTIVATING AGENTS

2016 ◽  
Vol 28 (2) ◽  
pp. 225 ◽  
Author(s):  
M. Suvá ◽  
N. G. Canel ◽  
D. F. Salamone
Keyword(s):  
Embryo Development ◽  
Polar Body ◽  
Reproductive Technologies ◽  
Mitogen Activated Protein Kinase ◽  
Cleavage Rate ◽  
Extrusion Rate ◽  
Second Polar Body ◽  
Polar Body Extrusion ◽  
Bovine Oocytes

Haploid activation of bovine oocytes is important for reproductive technologies such as intracytoplasmic sperm injection (ICSI) or somatic cell nuclear transfer (SCNT). Nevertheless, it is still a highly inefficient procedure. The aim of this work was to combine different activation drugs, known to have different targets along the activation cascade, to find a more effective activation protocol. Cumulus-oocyte complexes (COC) were aspirated from slaughtered ovaries and in vitro-matured (IVM) for 22 h. Oocytes were activated with 5 µM ionomycin (IO) for 4 min and then randomly allocated into 1 of the following treatments: 50 µM roscovitine (ROSC), 10 µg mL–1 cycloheximide (CHX), ROSC and 10 µM PD0325901 (ROSC/PD), or CHX and PD (CHX/PD) for 5 h; 15 µM dehydroleucodine (DHL) or DHL and ROSC (DHL/ROSC) for 3 h; DHL and CHX for 3 h followed by 2 h with CHX; 5-min exposure to 7% ethanol 4 h post-IO (ET); or ET followed by ROSC (ET-ROSC). Controls were IO followed by 3 h of exposure to 1.9 mM 6-DMAP with or without a previous 3-h culture in TCM-199 (3 h in DMAP and DMAP, respectively). Embryos were cultured in SOF medium. Pronuclear formation (PN) and second polar body extrusion (2PB) were assessed by 5 µg mL–1 propidium iodide oocyte staining, 17 h after IO. Activation was defined as the presence of at least 1 PN, and 2PB extrusion rate was calculated regardless of the nuclear stage. Data were analysed by Fisher’s Test (P < 0.05). Activation (Table 1) was similar in all groups, with the exception of ROSC/PD and ET-ROSC that were the highest and DHL that was the lowest. Although ROSC or CHX seemed to improve 2PB rate when combined with DHL, cleavage decreased significantly, suggesting DHL itself, or its combination with these drugs, negatively affects embryo development. Group ET showed activation rates comparable to other treatments, but it was not reflected on cleavage, suggesting that ET induces PN formation but it might be inefficient to trigger embryo development. Nevertheless, this observation was not made for ET-ROSC, as it showed a higher cleavage rate than ET and ROSC alone. The mitogen-activated protein kinase (MAPK) inhibitor PD showed different effects when combined with ROSC or CHX, despite that they both act on the mammary fat pad (MPF). In ROSC/PD, a slight improvement was observed on activation and cleavage rates compared with ROSC. Group CHX/PD resulted in a slightly higher 2PB percentage, but a lower activation percentage that derived in a significantly lower cleavage than CHX. In conclusion, ROSC and CHX were the most effective single treatments for haploid activation. Moreover, some combined treatments, namely DHL/ROSC and DHL/CHX, proved to be as effective or better at 2PB extrusion rate, which is the defining feature in haploid activation. Table 1.Activation, second polar body extrusion (2PB) and cleavage of bovine oocytes activated with ionomycin followed by single or combined activating agents1

313 EFFECT OF DURATION OF THE GROWING PHASE OF OVULATORY FOLLICLES IN SUPERSTIMULATED HEIFERS ON OOCYTE COMPETENCE AFTER IN VITRO FERTILIZATION

2013 ◽  
Vol 25 (1) ◽  
pp. 303
Author(s):  
F. C. F. Dias ◽  
D. Dadarwal ◽  
M. Honparkhe ◽  
G. P. Adams ◽  
R. J. Mapletoft ◽  
...  
Keyword(s):  
Embryo Development ◽  
Transvaginal Ultrasound ◽  
Animal Health ◽  
Prostaglandin F2α ◽  
Ovarian Response ◽  
Oocyte Quality ◽  
Cleavage Rate ◽  
Vitro Fertilization ◽  
Oocyte Competence

We tested the hypotheses that extending the duration of follicular growth by superstimulation increases oocyte competence, and that FSH starvation at the end of superstimulatory treatment decreases oocyte competence. Heifers were allocated randomly to short FSH duration (n = 8), FSH starvation (n = 8), or long FSH duration (n = 8) groups. Five to 8 days after ovulation, transvaginal ultrasound-guided follicle ablation was done to synchronize follicle wave emergence, and a progesterone-releasing device (CIDR; Pfizer Animal Health, New York, NY, USA) was placed intravaginally. Short FSH and FSH starvation groups were given 8 doses of FSH (Folltropin-V; Bioniche Animal Health Inc., Belleville, ON, Canada) IM, whereas the long FSH group was given 14 doses of FSH at 12-h intervals, starting from the day of wave emergence (Day 0). Prostaglandin F2α (PGF) was administered twice, 12 h apart, on Day 3 in the short FSH group and on Day 6 in the other 2 groups. In all heifers, the CIDR was removed at the time of the second PGF treatment; pLH (Lutropin-V; Bioniche Animal Health Inc.) was given IM 24 h after CIDR removal, and cumulus–oocyte complexes (COC) were collected 24 h after pLH treatment. The COC were matured in vitro (6 h) and fertilized (IVF), and the embryos were cultured for 10 days. At 12 h after pLH, the long FSH group had a greater number of ≥9 mm follicles than the FSH starvation and short FSH groups (25.4 ± 5.3 v. 11.0 ± 2.1 and 10.6 ± 2.3, respectively; P < 0.03). The long FSH group also had more expanded COC than the FSH starvation group (P < 0.001), but did not differ from the short FSH group (93, 54, and 74%, respectively). The FSH starvation group had a greater proportion (P < 0.0001) of partially expanded COC (32%) and poor quality oocytes (70%) than did the long (1 and 33%) and short (4 and 45%) FSH groups; oocyte quality did not differ between long and short FSH groups. At 48 h after IVF, the cleavage rate was lower in the FSH starvation group compared with the short and long FSH groups (35, 54, and 56%, respectively; P = 0.003). After 9 days in culture, embryo development (morula + blastocyst) in the FSH starvation group was lower than that in the long FSH group, (18 v. 37%; P = 0.04), but did not differ from that in the short FSH group (25%). After removal of the data of one heifer in the FSH starvation group that produced 52% of total embryos in that group (outlier), the Day 9 blastocyst rate was lower in the FSH starvation group than in the short and long FSH groups (2% v. 14 and 21%, respectively; P = 0.02). In conclusion, extending the standard superstimulation protocol by 3 days enhanced ovarian response to FSH treatment, but did not improve oocyte competence, whereas a period of FSH starvation after FSH treatment compromised oocyte quality and embryo development.

PSV-5 Relationship between sire conception rate, sperm motility, sperm SERPINA5 relative concentration, and in vitro produced embryos in dairy bulls

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 310-310
Author(s):  
Saulo Menegatti Zoca ◽  
Julie Walker ◽  
Taylor Andrews ◽  
Adalaide C Kline ◽  
Jerica J Rich ◽  
...  
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Relative Concentration ◽  
Early Embryo ◽  
Conception Rate ◽  
Cleavage Rate ◽  
Blastocyst Rate ◽  
Positive Correlations ◽  
Sire Conception Rate

Abstract Sire conception rate (SCR) is a field measure of fertility among bulls, but it can be influenced by several factors (Sperm transport, sperm-egg binding, early embryo development, etc). The objective of this study was to evaluate the relationship between SCR, sperm motility, SERPINA5 concentrations, and in vitro embryo development. Measurements were performed in 19 bulls with SCR values ranging from -7.7 to 4.45. For each bull, an aliquot of frozen-thawed semen was used for analyses of total (TMOT) and progressive (PROG) motility. Remaining semen was fixed with 2% formaldehyde, and concentration of SERPINA5 was determined by immunolocalization (antibody SERPINA5/Dylight405; PA5-79976-Invitrogen / ab201798-Abcam). Mean fluorescence intensity was determined in ~200 sperm heads/bull. Approximately 149 oocytes/bull were fertilized in vitro for embryo development analysis (cleavage and blastocyst rates). Statistical procedures were performed in SAS (9.4) using the procedures CORR for correlations (SCR, TMOT, PROG, SERPINA5, cleavage and blastocyst) and GLIMMIX for comparison of “field-fertility” (SCR divided in HIGH or LOW) and “field-embryo-fertility” (LOW-SCR sires were divided based on blastocyst rate (HIGH or LOW) resulting in two classifications; LOW-HIGH≥31% and LOW-LOW≤26%, respectively). There were positive correlations (P &lt; 0.05) between cleavage-blastocyst (r=0.50), SERPINA5-cleavage (r=0.48), and TMOT-PROG (r=0.76). Sire SCR was not associated with SERPINA5, TMOT, PROG, cleavage and blastocyst rate (P &gt; 0.52). Among LOW-SCR sires, LOW-LOW sires (-4.83±0.60) tended to have a better SCR score than LOW-HIGH (-6.18±0.42) sires (P = 0.08), but there were no differences (P &gt; 0.43) between LOW-HIGH, LOW-LOW, and HIGH sires for SERPINA5, TMOT, PROG, and cleavage. In conclusion, some LOW SCR sires have good embryo development indicating a different mechanism for their low SCR; however, these differences in SCR could not be explained by TMOT, PROG, SERPINA5, cleavage and blastocyst. There were, however, positive correlations between cleavage-blastocyst rate, and SERPINA5-cleavage rate.

scholarly journals Significance of disorders of proteomic profile of follicular fluid for predicting the effectiveness of in vitro fertilisation in women with infertility

2018 ◽  
Vol 99 (3) ◽  
pp. 496-503
Author(s):  
O S Zolotykh ◽  
S V Lomteva ◽  
K Yu Sagamonova
Keyword(s):  
Assisted Reproductive Technology ◽  
Follicular Fluid ◽  
Molecular Mechanisms ◽  
Reproductive Technologies ◽  
Reproductive Technology ◽  
In Vitro Fertilisation ◽  
Proteomic Profile ◽  
Technology Programs ◽  
Group 2

Aim. To study the proteomic profile of follicular fluid in patients with infertility in assisted reproductive technology programs. Methods. The study included women with infertility included in assisted reproductive technology programs: 15 women who had in vitro fertilisation which resulted in pregnancy (group 1) and 16 women with a negative result of this program (group 2). Fractionation of the follicular fluid samples was performed using the sets of special magnetic beads. Proteomic profiling was performed by tandem MALDI-mass-spectrometry. The anti-Müllerian hormone level was measured by ELISA. Results. The study revealed differences in the detectability of follicular fluid proteins with different regulatory properties in patients of groups 1 and 2. With the negative outcome of in vitro fertilisation, expression of a number of proteins involved in the processes of folliculogenesis, ovulation, selection of the dominant follicle, as well as proteins necessary for the development of the zygote and blastula was reduced in females' follicular fluid. Increased expression in women from group 2 was registered for proteins enhancing proteolytic reactions, cell apoptosis, including oocytes, which disrupt the positive action of activin and damage structural and functional state of mitochondria. A definite relationship was found between the level of anti-Müllerian hormone and rate of detection of a number of proteins, in particular protocadherin-2α, cystatin C, betaglycan, prostatic acid phosphatase, and dermicidin. Conclusion. The revealed changes in proteomic profile of the follicular fluid obviously play an important role in the molecular mechanisms that determine the effectiveness of assisted reproductive technologies; the identified differentially expressed proteins can serve as objective markers for predicting the outcomes of in vitro fertilisation.

scholarly journals Improvement of quality of oocytes collected in the autumn by enhanced removal of impaired follicles from previously heat-stressed cows

Reproduction ◽  
2001 ◽  
pp. 737-744 ◽  
Author(s):  
Z Roth ◽  
A Arav ◽  
A Bor ◽  
Y Zeron ◽  
R Braw-Tal ◽  
...  
Keyword(s):  
Heat Stress ◽  
Embryo Development ◽  
Treated Group ◽  
Oocyte Quality ◽  
Control Group ◽  
Delayed Effect ◽  
Cleavage Rate ◽  
Lactating Cows ◽  
Summer Heat

The fertility of dairy cows decreases during the summer and remains low during the cooler autumn although the animals are no longer under heat stress. The aim of this study was to characterize a delayed effect of summer heat stress on oocyte quality in the autumn and to improve oocyte quality by enhanced removal of follicles damaged during the previous summer. Lactating cows (n = 16) were subjected to heat stress during the summer. In autumn, ovarian follicles (3-7 mm in diameter) were aspirated by an ultrasound-guided procedure during four consecutive oestrous cycles. Follicles were aspirated from control cows on day 4 and from treated cows on days 4, 7, 11 and 15 of each oestrous cycle. All cows received PGF(2alpha) and GnRH injections on days 19 and 21, respectively, and maintained cyclicity, as indicated by plasma progesterone concentrations. On day 4 of each cycle, the oocytes recovered were examined morphologically, matured and activated in vitro, and cultured for 8 days. In cycle 1 (early October) both groups showed low percentages of grade 1 oocytes, cleavage, four- and eight-cell embryos, morulae and parthenogenetic blastocysts. Subsequently, the number of grade 1 oocytes increased earlier (cycle 2) in treated than in control cows (cycle 3; P < 0.05). The cleavage rate in the control group remained relatively low throughout (32-58%), whereas in the treated group it increased from 40% (cycle 1) to 75% (cycles 3 and 4; P < 0.05). The number at each stage of embryo development increased slightly but remained low throughout in the control group, whereas in the treated group significant (P < 0.05) increases of all stages were observed in cycles 3 and 4. The results show a delayed effect of summer heat stress on oocyte quality and embryo development in the autumn. Enhanced removal of the impaired cohort of follicles led to earlier emergence of healthy follicles and high quality oocytes in the autumn.

scholarly journals Influence of nutrition on the effectiveness of superovulation programmes in ewes: effect on oocyte quality and post-fertilization development

Reproduction ◽  
2003 ◽  
pp. 543-553 ◽  
Author(s):  
JM Lozano ◽  
P Lonergan ◽  
MP Boland ◽  
D O'Callaghan
Keyword(s):  
Embryo Development ◽  
Control Diet ◽  
Early Embryo ◽  
Low Energy ◽  
Lower Percentage ◽  
Uterine Tissue ◽  
Cleavage Rate ◽  
Ad Libitum

Two experiments were carried out to study the effect of nutrition on embryo development in two periods in superovulated ewes (Expt 1) and on oocyte developmental capacity during the late follicular phase (Expt 2). In Expt 1, a lower superovulation response in terms of animals ovulating (P < 0.05), ovulation rate per ewe ovulating (P = 0.1) and number of good quality embryos per animal treated (P < 0.07) was noted in ewes fed an ad libitum diet compared with ewes offered control (1.5 times the daily maintenance energy requirements, 1.5 x M) or low energy (0.5 x M) diets. Nutrition also modified the morphological and functional quality of the oocytes and embryos recovered. Thus, 92% of day 4 embryos recovered from ewes offered the control diet were classified as good embryos, compared with 70 and 82% of those recovered from ewes offered the ad libitum and low diets, respectively (P < 0.05). Ewes offered the ad libitum diet had a greater percentage of poorly developed embryos compared with ewes offered the control or low diets (P < 0.05). Ewes fed the low diet tended to have more non-fertilized oocytes than ewes offered the control diet (P = 0.09). Diet of recipient ewes to which good quality embryos were transferred on day 4 did not affect embryo quality, when assessed 12 days later (day 16 of pregnancy). However, recipient diet affected prostaglandin F(2alpha) (PGF(2alpha)) production in vitro, and uterine tissue that originated from recipient ewes on the low diet secreted more PGF(2alpha) relative to uterine tissue that originated from recipients on the control diet (P < 0.05). In Expt 2, fewer total (P < 0.05) and good quality (P < 0.01) oocytes and a lower percentage of good quality oocytes (P < 0.01) were obtained from superovulated ewes offered the ad libitum diet compared with ewes offered the low diet. In addition, cleavage rate tended to be higher (51 versus 35%, P = 0.09) in ewes offered the low diet compared with ewes offered the ad libitum diet. In conclusion, changes in diet can affect the quality of the oocyte and embryo in superovulated sheep. A lower superovulation response and a decrease in the quality of oocytes and embryos indicate that ad libitum diets are highly detrimental for superovulatory programmes when compared with low and control diets. In addition, the results from the present study indicate that a low energy diet during early embryo development increased the uterine production in vitro of PGF(2alpha) which could lead to a poor uterine environment thereby compromising the development of the embryo.

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