Structural modifications induced by an in vitro maturation process in zona pellucida glycoproteins of bovine oocytes. A Raman microspectroscopy analysis

RSC Advances ◽  
2016 ◽  
Vol 6 (86) ◽  
pp. 83429-83437 ◽  
Author(s):  
G. Rizo ◽  
M. Roldán-Olarte ◽  
D. C. Miceli ◽  
L. E. Jiménez ◽  
R. M. S. Álvarez
Keyword(s):  
Zona Pellucida ◽  
In Vitro Maturation ◽  
Raman Microspectroscopy ◽  
Maturation Process ◽  
Structural Modifications ◽  
Bovine Oocytes

Raman microspectroscopy is useful for discrimination between immature and in vitro matured bovine oocytes. Modifications in the glycoproteins of the zona pellucida exerted by the maturation methods might influence the process of in vitro production.

Zona pellucida birefringence correlates with developmental capacity of bovine oocytes classified by maturational environment, COC morphology and G6PDH activity

2012 ◽  
Vol 24 (4) ◽  
pp. 568 ◽  
Author(s):  
Eva Held ◽  
Eva-Maria Mertens ◽  
Abdollah Mohammadi-Sangcheshmeh ◽  
Dessie Salilew-Wondim ◽  
Urban Besenfelder ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Zona Pellucida ◽  
In Vitro Maturation ◽  
Immature Oocytes ◽  
Developmental Capacity ◽  
Bovine Oocytes ◽  
Scanning Electron

In the present study we aimed to analyse structural changes during in vitro maturation of the bovine zona pellucida (ZP) by scanning electron microscopy (SEM) ands zona pellucida birefringence (ZPB). Here we show that alterations during in vitro maturation invasively analysed by SEM are reflected in ZPB. In vivo-matured oocytes displayed significantly lower birefringence parameters and significantly higher blastocyst rates compared with in vitro-derived oocytes (39.1% vs 21.6%). The same was observed for in vitro-matured oocytes with cumulus–oocyte complex (COC) Quality 1 (Q1) compared with Q3-COCs with respect to zona birefringence and developmental capacity. Immature oocytes with Q1-COCs displayed higher ZPB values and a higher developmental capacity to the blastocyst stage (27.7% vs 16.9%) compared with immature Q3-COCs. Considering in vitro-matured oocytes, only those with Q1-COC showed a trend for ZPB similar to in vivo-matured oocytes. Therefore, a decreasing trend for ZPB during in vitro maturation seems to be typical for high-quality oocytes and successful cytoplasmic maturation. In accordance, fully-grown immature oocytes reached significantly higher blastocyst rates (32.0% vs 11.5%) and lower ZPB values compared with still-growing ones. In conclusion, we successfully evaluated the applicability of zona imaging to bovine oocytes: alterations during in vitro maturation invasively analysed by scanning electron microscopy were reflected in the birefringence of the zona pellucida of bovine oocytes affecting developmental capacity at the same value. Therefore ZPB measurement by live zona imaging has potential to become a new tool to assess correctness of in vitro maturation and to predict developmental competence.

160 EFFECT OF cAMP MODULATORS ON IN VITRO MATURATION OF BOVINE OOCYTES

2014 ◽  
Vol 26 (1) ◽  
pp. 193 ◽  
Author(s):  
S. E. Farmer ◽  
T. L. Adams ◽  
J. A. Sarmiento-Guzmán ◽  
C. L. Bailey ◽  
K. R. Bondioli
Keyword(s):  
In Vitro Maturation ◽  
Transvaginal Ultrasound ◽  
Inhibitory Effect ◽  
Maturation Process ◽  
In Vitro Production ◽  
Angus Cattle ◽  
Significant Difference ◽  
Chi Squared ◽  
Bovine Oocytes

The efficiency of in vitro production (IVP) has remained low due to an inadequate in vitro maturation (IVM) system. Thus far, the most promising methods of improving IVM utilise cAMP modulators to slow the maturation process by keeping cAMP high. This experiment compares standard IVM to an extended IVM as previously described by Albuz (2010 Hum. Reprod. 25, 2999–3011). The extended IVM consists of a 2-h pre-IVM phase with FSK (an adenylate cyclase activator) and 3-isobutyl-1-methylxanthine [IBMX, a nonspecific phosphodiesterase (PDE) inhibitor], followed by a 31-h IVM phase with cilostamide (a PDE3 inhibitor). 3-Isobutyl-1-methylxanthine inhibits all PDE in both the oocyte and CC, whereas cilostamide inhibits PDE3 only in the oocyte, allowing a gradual reversal of inhibition. Bovine oocytes (n = 363) were obtained by transvaginal ultrasound-guided aspiration of both Brahman and Angus cattle over 4 collection days. Oocytes from each cow were divided into 2 groups. The first group was placed in standard 23-h IVM medium composed of TCM-199 supplemented with 10% fetal bovine serum, sodium pyruvate, penicillin and streptomycin, glutamine, and FSH, and cultured in 5% CO2 at 39°C. A subset of oocytes was removed from maturation at 8, 13, 18, and 23 h. The second group was placed into a pre-IVM medium of HEPES-TALP supplemented with 100 μM FSK and 500 μM IBMX for 2 h at 39°C, and then moved into standard maturation medium supplemented with 20 μM cilostamide for 31 h (5% CO2, 39°C). Oocytes were sampled at 8, 13, 18, 23, 28, and 33 h. Oocytes were stained with 1% orcein and nuclear status was examined for each sample time (Table 1). Data were analysed using a chi-squared test. At 8 h, there was a significant difference (P < 0.001) between GV stage of IVM and extended IVM (7.1 v. 73.2%), and between metaphase I (MI) stage IVM and extended IVM (76.2 v. 4.9%). At the 23-h time sample, there was a significant difference between metaphase II (MII) IVM and MII extended IVM (78.9 v. 30.6%; P < 0.001). There was also a significant difference between MII oocytes at 23 h IVM and MII oocytes at 33 h extended IVM (78.9 v. 33.3%; P < 0.001). These results are consistent with the hypothesis that cAMP modulators slow the nuclear maturation process. However, results also suggest that the inhibitory effect of the cAMP modulators is not completely reversible. Oocytes appear to arrest at MI by 23-h extended IVM and do not progress to MII at the same rate as standard IVM. Table 1.Nuclear status of bovine oocytes after standard IVM and extended IVM with cAMP modulators

Development to the late morula or blastocyst stage following in vitro maturation and fertilization of bovine oocytes

1989 ◽  
Vol 18 (1-3) ◽  
pp. 139-148 ◽  
Author(s):  
Y. Fukui ◽  
M. Urakawa ◽  
C. Sasaki ◽  
N. Chikamatsu ◽  
H. Ono
Keyword(s):  
In Vitro Maturation ◽  
Blastocyst Stage ◽  
Bovine Oocytes
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