Relationship between chromatin configuration and in vitro maturation ability in guinea pig oocytes

It has been reported that immature, equine oocytes can be maintained in meiotic arrest at 24°C. To evaluate a commercial equine embryo holding medium for storage of equine oocyte at 24°C and to determine the effect of holding on maturation kinetics, cumulus‐oocyte complexes (COC) were recovered from slaughtered mares and placed in Syngro® Embryo Holding Solution at 22–25°C for 18–20 h (OH Group) or placed directly in DMEM-F12-based in vitro maturation (IVM) conditions (D-Mat Group) at 5% CO2 in air at 38.5°C. Maturation rate (metaphase II percentage; MII%) was assessed (presence of polar body under stereomicroscope) after denudation at 22, 24, and 28 h. After assessment, the denuded oocytes that were considered immature were placed back in IVM, reassessed at 24 and 28 h, and MII% was compared with that of oocytes remaining uninterrupted in IVM for 24 and 28 h. One-way ANOVA was used to compare dependent variable in different groups using PROC ANOVA (SAS, version 9.2, SAS Institute, Cary, NC, USA). A random selection of mature oocytes from both groups were fertilised using intracytoplasmic sperm injection (ICSI). A total of 250 injected oocytes were cultured in DMEM-F12 with 10% FCS. Blastocyst rates in OH and D-mat groups were similar (7.1% v. 6.3%). At 22 h, significantly more oocytes reached the MII stage in the OH group than in the D-Mat group, but MII% was similar in both groups at 24 and 28 h (Table 1). Denuded, immature oocytes reached similar maturation rate as the undenuded oocytes in the same group. Our data suggest that oocytes can be held in Syngro® Embryo Holding Solution at 22–25°C for 18–20 h without compromising oocyte developmental competence. Overnight holding of oocytes accelerates maturation with similar maturation rate at 22, 24, and 28 h of IVM in the OH group. Denudation of immature oocytes after 22 h of IVM and returning the denuded oocytes to IVM does not affect the progression of maturation. In subsequent experiment, overnight held oocytes were fixed and stained (Hoechst 33342) and MII% was evaluated after 20, 22, and 28 h of IVM. Chromatin configuration confirmed that stored oocytes reach the MII stage at 22 h. Maturation rates were significantly lower at 20 h, suggesting that 22 h of IVM is required for stored oocytes. Table 1.Maturation rates (% in MII stage) at 22, 24, and 28 h of IVM for equine oocytes held in Syngro® Embryo Holding Medium before IVM (OH) and oocytes placed directly in IVM (D-Mat) Thanks to I. Lemahieu and P. Van Damme. Study was supported by the Special Research Fund at UGent.

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132 BLASTOCYST DEVELOPMENT OF EQUINE OOCYTES WITH LOW MEIOTIC COMPETENCE HELD IN ROSCOVITINE BEFORE IN VITRO MATURATION

Reproduction Fertility and Development ◽

10.1071/rdv17n2ab132 ◽

2005 ◽

Vol 17 (2) ◽

pp. 216

Author(s):

Y.H. Choi ◽

L.B. Love ◽

D.D. Varner ◽

K. Hinrichs

Keyword(s):

Epithelial Cells ◽

In Vitro Maturation ◽

Developmental Competence ◽

Blastocyst Development ◽

Exact Test ◽

Nuclear Maturation ◽

Significant Difference ◽

Chromatin Configuration ◽

Meiotic Competence

At the time of recovery, immature equine oocytes may be separated into those with either expanded cumuli (Ex) or compact cumuli (Cp). The Cp oocytes originate from viable follicles but are largely juvenile, with low meiotic competence (20 to 30% maturation to MII), and possibly reduced developmental competence. We previously found that in Cp oocytes recovered immediately after slaughter, suppression of meiosis with roscovitine for 24 h before maturation increased embryo development at 4 days after intracytoplasmic sperm injection (ICSI; Franz et al. 2003 Reproduction 125, 693–700). The present study was conducted to evaluate the effect of roscovitine suppression on nuclear maturation and blastocyst formation of Cp oocytes recovered after transport of ovaries from the abattoir (i.e. recovered 5–9 h after slaughter). Compact oocytes recovered from transported ovaries were cultured in M199 with 10% FBS containing 66 μM roscovitine with or without an oil cover. After 16–18 or 24 h, oocytes were fixed to examine the chromatin configuration. Treatment for 16–18 h without oil resulted in the lowest rate of meiotic resumption (0%); thus this treatment was utilized in further studies. Resumption in other treatments ranged from 3 to 6%. Following roscovitine suppression, oocytes were cultured for 30 h in M199 with 10% FBS and 5 μU mL−1 FSH for maturation; control oocytes were cultured for 30 h in the same medium immediately after recovery. Mature oocytes were subjected to ICSI, then cultured in DMEM/F-12 with 10% FBS with or without co-culture with equine oviductal epithelial cells under mineral oil in 5% CO2 in air at 38.2°C, and then evaluated at 7.5 days. Progression to MII (82/376, 22%) after maturation of roscovitine-treated oocytes was similar to that for control oocytes (74/395, 19%). There was no significant difference in cleavage rates after ICSI (72–78%) among treatments. Development to blastocyst was highest in roscovitine-treated oocytes in DMEM/F-12 with co-culture (11/30, 37%); this was significantly higher than that of non-treated oocytes in DMEM/F-12 alone (5/36, 14%), but similar to that of non-treated/DMEM/F-12/co-culture (10/37, 27%) and roscovitine/DMEM/F-12 alone (8/39, 21%). These data indicate that roscovitine induces a fully reversible meiotic suppression in Cp equine oocytes recovered 5–9 h after slaughter, and that this suppression does not harm subsequent developmental competence. This treatment may be used to manipulate the time of onset of maturation of equine oocytes for ease of subsequent procedures. Co-culture with oviductal epithelial cells tended to increase blastocyst rate (P = 0.1, Fisher's exact test) in contrast to our previous findings with embryos from Ex oocytes (Choi et al. 2004 Biol. Reprod. 70, 1231–1238). Further work is needed to determine whether this is related to differences in intrinsic developmental competence between oocyte types. This work was supported by the Link Equine Research Endowment Fund (Texas A&M University).

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Toxicity effect of Auxemma oncocalyx fraction and its active principle oncocalyxone A on in vitro culture of caprine secondary follicles and in vitro oocyte maturation

Semina Ciências Agrárias ◽

10.5433/1679-0359.2017v38n3p1361 ◽

2017 ◽

Vol 38 (3) ◽

pp. 1361 ◽

Cited By ~ 3

Author(s):

Johanna Leiva-Revilla ◽

Jesús Cadenas ◽

Luis Alberto Vieira ◽

Claudio Cabral Campello ◽

Juliana Jales de Hollanda Celestino ◽

...

Keyword(s):

Growth Rate ◽

In Vitro Culture ◽

Oocyte Maturation ◽

In Vitro Maturation ◽

Cellular Proliferation ◽

Control Group ◽

Antral Follicles ◽

Chromatin Configuration ◽

In Vitro Oocyte Maturation

Crude extract of the heartwood of Auxemma oncocalyx (A. oncocalyx) and its main component i.e., Oncocalyxone A (onco A), have elevated antioxidant and anti-tumoral activity, but studies on the action of these drugs regarding folliculogenesis are lacking. The aim of this study was to evaluate the effect of A. oncocalyx and onco A on the in vitro culture of isolated secondary follicles and on the in vitro maturation of oocytes from caprine antral follicles grown in vivo. Isolated secondary follicles were randomly distributed in six groups; the non-cultured control was immediately fixed upon isolation. The remaining follicles were cultured for 7 days in ?-MEM+ alone (control) or supplemented with DMSO, doxorrubicin, A. oncocalyx or onco A. After culture, follicles were evaluated for antrum formation, growth rate, apoptosis (TUNEL) and cellular proliferation (PCNA), as well as gene expression of Bcl2 and Bax. Additionally, cumulus oocyte complexes (COCs) were aspirated and allocated into five treatments for in vitro maturation: control, cultured only in maturation base medium (TCM 199+); or supplemented with DMSO; DXR; A. oncocalyx or onco A. After in vitro maturation, oocyte chromatin configuration and viability were assessed. After 7 days of culture, there was a reduction (P < 0.05) in the percentage of morphologically intact follicles, antrum formation, growth rate and number of PCNA positive granulosa cells in DXR treatment compared to the other treatments. In the DXR treatment a higher percentage (P < 0.05) of TUNEL positive follicles and higher (P < 0.05) relative BAX:BCL2 mRNA ratio’s were observed. After in vitro maturation of the COCs DXR, A. oncocalyx and onco A treatments had a greater (P < 0.05) percentage of abnormal oocytes and a lower (P < 0.05) percentage of viable oocytes as compared with the control group. However, only DXR and onco A treatments increased (P < 0.05) the percentage of alive oocytes with abnormal chromatin configuration. There were no differences in maturation rates between the control group and DXR, A. oncocalyx and onco A treatments. In conclusion, under our culture conditions, A. oncocalyx and onco A do not exhibit a toxic effect on isolated secondary follicles and on maturation rates of COCs recovered from antral follicles, however, these drugs negatively affected the COCs viability. Thus, the use of culture biotechnologies as an in vitro secondary follicle culture and in vitro oocyte maturation toxicity testing are appropriated methods to evaluate the possible effects of drugs in folliculogesis.

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In vitro maturation of primate oocytes retrieved at the time of cesarean section

Journal of the Society for Gynecologic Investigation ◽

10.1016/s1071-5576(97)86351-4 ◽

1998 ◽

Vol 5 (1) ◽

pp. 106A-106A

Author(s):

R BRZYSKI ◽

M LELAND ◽

C EDDY

Keyword(s):

Cesarean Section ◽

In Vitro Maturation

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Effect of isoflavone compounds on the in vitro maturation of sheep oocytes

Planta Medica ◽

10.1055/s-0035-1565604 ◽

2015 ◽

Vol 81 (16) ◽

Author(s):

AA Amir ◽

GB Martin ◽

JM Kelly ◽

DO Kleemann ◽

Z Durmic ◽

...

Keyword(s):

In Vitro Maturation

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Inhibition of Human and Animal Platelet Adhesiveness to Glass Bead Columns by Adenosine, Dipyridamole, Chlorpromazine and Acetylsalicylic Acid

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648055 ◽

1976 ◽

Vol 36 (02) ◽

pp. 401-410 ◽

Cited By ~ 6

Author(s):

Buichi Fujttani ◽

Toshimichi Tsuboi ◽

Kazuko Takeno ◽

Kouichi Yoshida ◽

Masanao Shimizu

Keyword(s):

Guinea Pig ◽

Acetylsalicylic Acid ◽

Guinea Pigs ◽

Glass Bead ◽

Animal Species ◽

Inhibitory Effect ◽

Rabbit Platelet ◽

Platelet Adhesiveness

SummaryThe differences among human, rabbit and guinea-pig platelet adhesiveness as for inhibitions by adenosine, dipyridamole, chlorpromazine and acetylsalicylic acid are described, and the influence of measurement conditions on platelet adhesiveness is also reported. Platelet adhesiveness of human and animal species decreased with an increase of heparin concentrations and an increase of flow rate of blood passing through a glass bead column. Human and rabbit platelet adhesiveness was inhibited in vitro by adenosine, dipyridamole and chlorpromazine, but not by acetylsalicylic acid. On the other hand, guinea-pig platelet adhesiveness was inhibited by the four drugs including acetylsalicylic acid. In in vivo study, adenosine, dipyridamole and chlorpromazine inhibited platelet adhesiveness in rabbits and guinea-pigs. Acetylsalicylic acid showed the inhibitory effect in guinea-pigs, but not in rabbits.

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INHIBITION OF LIPOLYTIC ACTIVITY OF SYNTHETIC β1–24 AND β1–39 CORTICOTROPHIN BY ANTI-ACTH ANTISERUM

Acta Endocrinologica ◽

10.1530/acta.0.0510088 ◽

1966 ◽

Vol 51 (1) ◽

pp. 88-94 ◽

Cited By ~ 1

Author(s):

A. Villanueva ◽

S. J. H. Ashcroft ◽

J. P. Felber

Keyword(s):

Guinea Pig ◽

Lipolytic Activity ◽

Peptide Hormones ◽

Fat Pad ◽

Double Antibody ◽

Epididymal Fat ◽

Antibody Complex ◽

Radio Immunoassay ◽

Antigen Antibody

ABSTRACT The synthetic ACTH peptides β1–39 and β1–24 stimulated lipolysis as determined by the rat epididymal fat pad in vitro. The stimulating effect of these peptides was diminished by prior incubation of the peptides with antibodies produced by the guinea-pig against ACTH. The stimulating effect of these hormones was also diminished by the double antibody system used in the radio-immunoassay of ACTH and other peptide hormones, in which incubation with antiserum is followed by precipitation of the antigen-antibody complex by rabbit anti-guinea-pig-γ-globulin.

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