The rôle of the germinal vesicle in oöcyte maturation in anurans as revealed by the removal and transplantation of nuclei

Development ◽  
1964 ◽  
Vol 12 (4) ◽  
pp. 851-873
Author(s):  
T. A Dettlaff ◽  
L. A. Nikitina ◽  
O. G. Stroeva
Keyword(s):  
Embryonic Development ◽  
Sea Water ◽  
Germinal Vesicle ◽  
Maturation Stages ◽  
Cytoplasmic Maturation ◽  
Cerebratulus Lacteus

A Study of nucleo-cytoplasmic interactions in pre-embryonic development, at maturation stages in particular, is of great interest. During this period oöplasmic segregation takes place, specific properties of the cortex arise, and the cytoplasm obtains information from the oöcyte nucleus which promotes the development of the embryo during all the changes related to fertilization, cleavage and blastulation (Wilson, 1925; Raven, 1961; Brachet, 1957, 1960; Briggs & King, 1959; Neyfakh, 1962). The first data on the rôle of the oöcyte nucleus (germinal vesicle or GV) in cytoplasmic maturation were obtained by Delage (1899, 1901) on the starfish Asterias glacialis, and by Wilson (1903) on the nemertine Cerebratulus lacteus. In both species oöcytes are expelled by the female before the GV dissolves, and mature in sea water. This allowed Delage and Wilson to experiment with oöcytes at different maturation stages.

Action of actinomycin and puromycin upon frog oocyte maturation

Development ◽  
1966 ◽  
Vol 16 (1) ◽  
pp. 183-195
Author(s):  
T. A. Dettlaff
Keyword(s):  
Oocyte Maturation ◽  
Germinal Vesicle ◽  
Cell Types ◽  
Follicle Cells ◽  
Oocyte Nucleus ◽  
Frog Oocyte ◽  
Cytoplasmic Maturation ◽  
Gonadotropic Hormones ◽  
Cerebratulus Lacteus

In amphibians, as well as in other vertebrates, occytes start to mature under the action of hypophyseal gonadotropic hormones. (In this paper the term ‘maturation’ implies the transformation into ripe eggs of oocytes that have finished growing.) In the course of maturation the oocytes themselves and the follicle cells surrounding them undergo changes; the changes of these two cell types are not causally connected, their coincidence in time is easily broken in unfavourable conditions (Wright, 1945; Tchou-Su & Wang Yu-lan, 1958). As shown earlier (Dettlaff, Nikitina & Stroeva, 1964), hypophyseal gonadotropic hormones affect amphibian oocytes through the oocyte nucleus, the germinal vesicle. As a result of their action the nuclear sap of the germinal vesicle acquires the ability to induce cytoplasmic maturation (the property revealed by Delage on the oocytes of Asterias glacialis, 1899, 1901, and by Wilson on those of Cerebratulus lacteus, 1903).

320 MPF ACTIVITY AND DEVELOPMENTAL COMPETENCE IN DIFFERENT SIZES OF PREPUBERTAL GOAT OOCYTES

2006 ◽  
Vol 18 (2) ◽  
pp. 267 ◽  
Author(s):  
B. Anguita ◽  
A. R. Jimenez-Macedo ◽  
D. Izquierdo ◽  
M. T. Paramio
Keyword(s):  
Embryonic Development ◽  
Kinase Activity ◽  
Developmental Competence ◽  
Kinase Assay ◽  
Oocyte Diameter ◽  
Cdc2 Kinase ◽  
Cell Extracts ◽  
Morphological Criteria ◽  
Blastocyst Rate ◽  
Cytoplasmic Maturation

Developmental competence of prepubertal goat oocytes recovered from a slaughterhouse is low, probably due to an incomplete cytoplasmic maturation. Regulation of cytoplasmic maturation is still unknown, although maturation-promoting factor (MPF) is suggested to play an important role in this process. To better understand the role of MPF in cytoplasmic maturation, we have studied MPF kinase activity in oocytes with different developmental competence. Ovaries were obtained from a slaughterhouse, and oocytes were recovered by slicing and were selected according to morphological criteria. Some oocytes were denuded and classified in diameter groups (<110 μm, 110–125 μm, 125–135 μm, and >135 μm), placed in lysis buffer (50 mM Tris-HCl (pH 7.5), 0.5 M NaCl, 5 mM EDTA, 0.01% Brij35, 1 mM phenyl methyl sultonyl fluoride (PMSF), 0.05 mg/mL leupeptin, 50 mM 2-mercaptoethanol, 25 mM α-glycerophosphate, 1 mM Na-orthovanadate) and frozen in liquid N2. Cell extracts were stored at −80°C until use. The rest of oocytes were matured in vitro in medium TCM199 supplemented with hormones, 10% (DBS), and 400 μM cysteamine, for 27 h in 5% CO2 in air and 38.5°C. After IVM, a sample of oocytes were also denuded, classified by diameters, and frozen as described above. The rest of oocytes were used for IVF in mDM with spermatozoa capacitated with heparin and ionomicin. After 24 h, presumptive zygotes were cultured for 7 days in medium SOF in 5% CO2, 5% O2, and 90% N2 at 38.5°C. At 48 h post-insemination, we added 0.1 μL FBS per embryo. Embryonic development was evaluated with Hoechst staining after IVC. MPF kinase activity was detected using the MESACUP cdc2 kinase assay kit (MBL Woburn, MA, USA). Briefly, the oocyte extract corresponding to 10 oocytes was mixed with 10× reaction buffer (25 mM HEPES buffer (pH 7.5), 10 mM MgCl2) and 10% biotinylated MV peptide (SLYSSPGGAYC). We added 0.1 mM ATP to start the reaction. The mixture was incubated at 30°C for 30 min. The reaction was finished by adding 200 μL of PBS containing 50 mM EGTA. The phosphorylated MV peptide was detected by specific antibody using an ELISA procedure, and the OD was measured at 492 nm. Fisher's exact test was used to analyze IVC results, and ANOVA to analyze cdc2 kinase activity results. We considered differences statistically significant when P < 0.05. Results are shown in Table 1. We observed that embryonic cleavage and blastocyst rate increased with oocyte diameter. The MPF activity detected was higher in the largest oocytes after IVM. As a consequence, we could establish that oocytes with a higher MPF activity are more capable of maintaining embryonic development until the blastocyst stage, which may indicate the important role that MPF plays in cytoplasmic maturation. Table 1. Cleavage, blastocyst rate, and MPF kinase activity in different sizes of prepubertal goat oocytes

336 MEIOTIC DEVELOPMENT AND CORTICAL GRANULES DISTRIBUTION IN CANINE OOCYTES DURING IN VITRO MATURATION

2010 ◽  
Vol 22 (1) ◽  
pp. 324 ◽  
Author(s):  
M. De los Reyes ◽  
D. Luna ◽  
J. Palomino
Keyword(s):  
In Vitro Maturation ◽  
Germinal Vesicle ◽  
Fluorescein Isothiocyanate ◽  
Cumulus Cells ◽  
Homogeneous Distribution ◽  
Cortical Granules ◽  
Dapi Staining ◽  
Nuclear Maturation ◽  
Cytoplasmic Maturation

Low development of IVM canine oocytes could be in part attributed to an impaired cytoplasmic maturation. In mammalian oocytes, migration and the redistribution of cortical granules (CGs) around the periphery of the oocyte contribute to the inhibition of polyspermy and it is an important criterion to evaluate cytoplasmic maturation. The state of nuclear maturation and the distribution of CGs were evaluated in canine oocytes cultured for different periods in order to compare the synchrony of nuclear and cytoplasmic maturation during in vitro maturation. Bitch ovaries at different stages of the estrous cycle were obtained following ovariectomy. COCs with compact cumulus cells showing a homogeneous cytoplasm were selected for experiments. Thirty-six COCs were processed at immature stage, placed in PBS medium until evaluation. A total of 275 COCs were matured in vitro for 48, 72, and 96 h in TCM-199 with Earle’s salt supplemented with 25 mM Hepes, 10% FCS, 0.25 mM pyruvate, 10 IU mL-1 of hCG, 300 IU mL-1 penicillin, and 20 mg mL-1 streptomycin, at 38.5°C and 5% CO2. At each culture period, the oocytes were stained with Lens culinaris agglutinin (LCA), labeled with fluorescein isothiocyanate, and the CGs distributions were examined under a fluorescent microscope. The nuclear status of the denuded oocytes was determined by DAPI staining under a fluorescence microscope. For each treatment, at least four replicates were performed and the data was analyzed by ANOVA using Tukey’s test to determine the differences P < 0.05. Three types of CGs distribution were distinguished during canine oocyte maturation: (1) homogeneous distribution throughout the cytoplasm including the cortex; (2) heterogeneous (clusters) within the cytoplasm and (3) densely distributed beneath the oolemma. Nuclear stages were classified as immature or germinal vesicle (GV) stage; resumption of meiosis or germinal vesicle break down (GVBD); metaphase I to telophase I (MI toTel I); and mature or second metaphase (MII). The distribution patterns of GCs were different (P < 0.05) among oocytes cultured for different periods and the nuclear maturation status also differed between oocytes cultured for different intervals (P < 0.05). Most (>84%) of the immature oocytes at GV showed a uniform distribution of CGs throughout the cytoplasm. At 48 h of culture, CGs distribution was mainly Type 2 (25%) and 3 (61%) and the oocytes were at GVBD (33%) and MI-Tel I (33%) stages. Most nuclei of the type 3 oocytes were in the MI (40%) and MII (11%) stages, corresponding to those oocytes matured for 72 (88%) or 96 h (71%). These results indicate that canine oocytes migrate to the cortex during IVM and this process is not finished before 72 h of culture. In addition, although the re-distribution of the CGs occurred in parallel with nuclear maturation, the oocytes cannot always proceed to the MII stage; however, in such oocytes the CGs are distributed beneath the oolemma. Supported by Grant FONDECYT 1080618.

scholarly journals Enucleolation and nucleolus transfer in mammalian oocytes and zygotes

2019 ◽  
Vol 63 (3-4-5) ◽  
pp. 253-258
Author(s):  
Michal Benc ◽  
Josef Jr. Fulka ◽  
Franti&scaron;ek Strej&ccaron;ek ◽  
Martin Morovi&ccaron; ◽  
Matej Murín ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Oocyte Maturation ◽  
Real Function ◽  
Morphological Structure ◽  
Germinal Vesicle ◽  
Early Embryonic Development ◽  
The Real ◽  
Germinal Vesicles

The oocyte GV/GVs (germinal vesicle/germinal vesicles) and zygot PN/PNs (pronucleus/pronuclei) of some mammals contain clearly visible nucleoli which exhibit an atypical morphological structure. These nucleoli (NCLs) can be relatively easily manipulated, i.e. removed from GVs/PNs or eventually transferred into another oocyte/zygote. Thus, with the help of micromanipulation techniques it was possible to uncover the real function(s) they play in processes of oocyte maturation and early embryonic development. The purpose of our review is to describe briefly the micromanipulation techniques that can be used for oocyte/zygote nucleoli manipulation. Moreover, we present some examples of results that were obtained in nucleolus manipulation experiments.

scholarly journals Porcine follicular fluid derived from > 8 mm sized follicles improves oocyte maturation and embryo development during in vitro maturation of pigs

Zygote ◽  
2020 ◽  
pp. 1-6
Author(s):  
Ji-Eun Park ◽  
Sang-Hee Lee ◽  
Yong Hwangbo ◽  
Choon-Keun Park
Keyword(s):  
Embryonic Development ◽  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Developmental Competence ◽  
Control Group ◽  
Cumulus Expansion ◽  
Treatment Groups ◽  
Nuclear Maturation ◽  
Cytoplasmic Maturation

Summary The aim of the present study was to investigate the effects of porcine follicular fluid (pFF) from large-sized (LFF; >8 mm in diameter) and medium-sized (MFF; 3–6 mm in diameter) follicles on the maturation and developmental competence of porcine oocytes. Cumulus–oocyte complexes (COCs) were collected from follicles 3–6 mm in diameter. The collected COCs were incubated for 22 h with LFF or MFF (in vitro maturation (IVM)-I stage) and were incubated subsequently for 22 h with LFF or MFF (IVM-II stage). Cumulus expansion was confirmed after the IVM-I stage and nuclear maturation was evaluated after the IVM-II stage. Intracellular glutathione (GSH) and reactive oxygen species (ROS) levels were measured and embryonic development was evaluated. Relative cumulus expansion and GSH levels were higher in the LFF group compared with in the MFF group after the IVM-I stage (P < 0.05). After the IVM-II stage, the numbers of oocytes in metaphase-II were increased in the LFF group and GSH content was higher in all of the LFF treatment groups compared with in the MFF treatment groups during both IVM stages (P < 0.05). ROS levels were reduced by LFF treatment regardless of IVM stage (P < 0.05). Blastocyst formation and the total numbers of cells in blastocysts were increased in all LFF treatment groups compared with the control group (P < 0.05). These results suggested that pFF from large follicles at the IVM stage could improve nucleic and cytoplasmic maturation status and further embryonic development through reducing ROS levels and enhancing responsiveness to gonadotropins.

Quantitative analysis of the intensity of zona pellucida birefringence of oocytes during IVF cycles

2010 ◽  
Vol 22 (8) ◽  
pp. 1167 ◽  
Author(s):  
Junping Cheng ◽  
Li Huang ◽  
Bing He ◽  
Fenghua Lu ◽  
Xiaoli Wang ◽  
...  
Keyword(s):  
Retrospective Study ◽  
Zona Pellucida ◽  
Embryo Quality ◽  
Imaging System ◽  
Germinal Vesicle ◽  
Clinical Results ◽  
Maturation Stage ◽  
System A ◽  
Maturation Stages ◽  
Cycle Outcome

To determine whether the age of the female patient, oocyte maturation stage, pronuclei, embryonic quality and/or cycle outcome are related to the intensity of the zona pellucida birefringence (ZPB) in oocytes during IVF procedures, a retrospective study was conducted on 214 IVF cycles using a polarisation imaging system. A negative correlation was found between ZPB intensity and the age of female patients (r = –0.44; P < 0.0001). For oocytes at different maturation stages, the ZPB score was higher at the germinal vesicle stage (20.77 ± 1.88) than at MI (10.99 ± 1.35; P < 0.001), and higher at MI than at MII (7.91 ± 0.65; P < 0.05). For the pronuclei (PN) of fertilised oocytes, the ZPB score of oocytes with 3PN was significantly higher than that of oocytes with 1PN or 0PN (7.94 ± 0.78 v. 5.57 ± 1.51 and 4.45 ± 0.85, respectively; P < 0.001). However, there were no differences in ZPB scores between oocytes with 2PN and 3PN. Neither embryo quality on Day 3 nor the clinical results could be predicted on the basis of the ZPB score (P > 0.05).

Coordination of nuclear and cytoplasmic oocyte maturation in eutherian mammals

1996 ◽  
Vol 8 (4) ◽  
pp. 485 ◽  
Author(s):  
JJ Eppig
Keyword(s):  
Growth Phase ◽  
Germinal Vesicle ◽  
Preimplantation Development ◽  
Preimplantation Embryos ◽  
Maternal Factors ◽  
Cell Stage ◽  
Nuclear Maturation ◽  
Cytoplasmic Maturation ◽  
Developmental Programme ◽  
Critical Aspects

As oocytes near the end of their growth phase, they become competent to undergo two aspects of maturation, cytoplasmic and nuclear. Both are essential for the formation of an egg having the capacity for fertilization and development to live offspring. Nuclear maturation encompasses the processes reversing meiotic arrest at prophase I and driving the progression of meiosis to metaphase II. Cytoplasmic maturation refers to the processes that prepare the egg for activation and preimplantation development. This review focuses on the developmental programmes whereby oocytes at the germinal vesicle (GV) stage acquire competence to undergo nuclear and cytoplasmic maturation, the coordination of programmes regulating the acquisition of these competencies in GV-stage oocytes, and the coordination of the maturational processes themselves. Although the developmental programme of the GV-stage oocyte for acquiring competence to complete preimplantation development does not appear to be tightly linked to the acquisition of competence to complete nuclear maturation, GV breakdown (GVB) is probably essential for activating some critical aspects of cytoplasmic maturation, particularly those related to fertilization and activation. Nuclear and cytoplasmic maturation are normally coordinated by this mechanism requiring the mixing of the GV contents with the cytoplasm at the time of GVB, but some processes of cytoplasmic maturation related to successful preimplantation development probably still occur without coordination with nuclear maturation. Thus, continued differentiation of GV-stage oocytes is necessary after the acquisition of competence to undergo nuclear maturation, to allow for the deposition of the maternal factors required for the development of preimplantation embryos beyond the 2-cell stage.

scholarly journals Effect of Temperature on Ovarian Maturation Stages and Embryonic Development of Mud Spiny Lobster, Panulirus polyphagus

2013 ◽  
Vol 9 (1) ◽  
pp. 1-13 ◽  
Author(s):  
M. Ikhwanuddi ◽  
S.N. Fatihah ◽  
A.H. Nurfaseha ◽  
M. Fathiah ◽  
M. Effenddy ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Spiny Lobster ◽  
Effect Of Temperature ◽  
Ovarian Maturation ◽  
Maturation Stages

253 OOCYTE PREMATURATION IN THE PRESENCE OF MILRINONE IMPROVES NUCLEAR BUT NOT CYTOPLASMIC MATURATION OF MACAQUE OOCYTES

2011 ◽  
Vol 23 (1) ◽  
pp. 224 ◽  
Author(s):  
E. C. Curnow ◽  
J. P. Ryan ◽  
D. M. Saunders ◽  
E. S. Hayes
Keyword(s):  
In Vitro Maturation ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Molecular Probes ◽  
Developmental Competence ◽  
Oocyte Growth ◽  
North West ◽  
Significant Difference ◽  
Cytoplasmic Maturation

During oocyte growth chromatin configuration of the germinal vesicle (GV) oocyte undergoes modification in relation to changes in transcriptional activity crucial for conferring meiotic as well as developmental competence on the oocyte. In the macaque oocyte, there are 3 distinct GV states: GV1, noncondensed chromatin; GV2, an intermediate state; and GV3, condensed chromatin. The aim of this study was to test the effects of a prematuration culture (PMC) system, using the phosphodiesterase type 3 inhibitor milrinone (MIL), on the synchronization of GV chromatin to the GV3 stage and assess metaphase II (MII) oocyte reduced glutathione (GSH) content as a measure of cytoplasmic maturation. Reagents were purchased from Sigma (St. Louis, MO, USA) unless stated otherwise. To assess the effect of PMC on GV chromatin status, immature oocytes retrieved from unstimulated ovaries were either fixed (2% paraformaldehyde+0.1% Triton-X100) immediately after follicular aspiration (t = 0) or after culture in a humidified atmosphere of 6% CO2 in air at 37°C for 24 h in modified Connaught Medical Research Laboratories medium (mCMRL) supplemented with 10% FCS (Hyclone, Logan, UT, USA) and 12.5 μM MIL in the absence (MILNil) or presence of 1.0 IU of FSH (MILFSH). For chromatin assessment, fixed GV oocytes were stained with 5 μg mL–1 of 4′,6-diamidino-2-phenylindole (Molecular Probes, Leiden, the Netherlands) and imaged using confocal microscopy. Following PMC, MILFSH oocytes were transferred to fresh mCMRL+FCS supplemented with 1.0 IU of recombinant human FSH and 1.0 IU of hLH and cultured for a further 30 h. Control and MILFSH oocytes were denuded of cumulus cells and assessed for maturation. The MII oocytes were prepared for GSH analysis, and total GSH content was determined using a commercial 5,5′-dithio-bis(2-nitrobenzoic acid) (DTNB)-GSH reductase recycling assay kit (North-West Life Science). The MII rates were compared using chi-square. Differences in oocyte GSH content were compared using t-test. Significant differences were determined at P < 0.05. There was no significant difference in the proportion of oocytes remaining at the GV stage following 24 h of PMC in MILNil or MILFSH (42/44, 96% v. 32/35, 91%, respectively). However, there was a significant reduction in GV1 chromatin (15/49, 31% v. 28/54, 52% and 22/58, 38%) and a significant increase in GV3 chromatin (23/49, 47% v. 14/54, 26% and 16/58, 28%) observed in MILFSH oocytes compared with both MILNil and t = 0 oocytes, respectively. The MII rate of MILFSH oocytes following in vitro maturation was significantly higher compared with the MII rate of control in vitro matured oocytes (91/167, 55% v. 83/243, 34%). There was no significant difference in the GSH content of GV oocytes from the time of oocyte collection (t = 0) or GV oocytes following PMC in MILFSH (3.69 ± 0.16 and 4.14 ± 0.28 pmol/oocyte, n = 39–49 oocytes). The GSH content of control in vitro matured MII oocytes was significantly greater than that of MILFSH-treated MII oocytes (3.13 ± 0.16 v. 2.02 ± 0.04 pmol/oocyte, n =53–54 oocytes). The PMC supported high rates of nuclear maturation, but cytoplasmic maturation, assessed by GSH content, was negatively affected. Further assessment following fertilization and development is required to determine the practical utility of PMC in a primate in vitro maturation setting.

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