In Vitro Development of in Vitro-Matured Porcine Oocytes Following Chemical Activation or in Vitro Fertilization1

Funahashi, Hiroaki; Cantley, T.C.; Stumpf, T. T.; Terlouw, Steven L.; Day, Billy N.

doi:10.1095/biolreprod50.5.1072

Cited by 112 publications

(71 citation statements)

References 36 publications

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“…In bovine oocytes, treatment with A23187 alone induces a decrease in the levels of MPF activity within 30 min [118]. Meiotic resumption and pronuclear formation have been observed in hamster [143,144], cattle [131,139] and pig [126,132,142,145,146] oocytes f o l l o w i n g t r e a t m e n t w i t h A 2 3 1 8 7 , w i t h parthenogenetic development, as assessed by cleavage and blastocyst formation, also being reported [118,126,142,145]. The concentration of A23187 and the duration of exposure to A23187 affect the efficiency of oocyte activation [131,142].…”

Section: Calcium Ionophoresmentioning

confidence: 99%

Calcium Release at Fertilization: Artificially Mimicking the Oocyte's Response to Sperm.

Grupen¹,

Nottle²,

Nagashima

2002

Journal of Reproduction and Development

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Abstract. The mechanism of sperm-induced calcium release has been the subject of many studies since the development in the late 1950s of in vitro culture systems that support mammalian fertilization. Despite efforts to elucidate the nature of the signal from the sperm that triggers both the early and late events of oocyte activation, the precise mechanism remains unresolved. Now, with the advent of somatic nuclear transfer technologies, the need to better understand this unique process has been recognised. Nuclear transfer embryos must be induced to commence development artificially because the activating signal from the sperm is absent. The primary activating stimulus is a large increase in the concentration of intracellular-free calcium and numerous physical and chemical treatments have been found to induce calcium changes that initiate the events of oocyte activation. Although live cloned offspring have been produced in a number of species, the overall efficiencies of the nuclear transfer procedures described thus far are unacceptably low and phenotypic anomalies are common. With the aim of improving these efficiencies, researchers are developing artificial activation treatments which induce oocyte responses that mimic those induced by fertilizing sperm. One strategy is to replicate the pattern of calcium change more closely. Another strategy is to couple an activating stimulus with treatments that inhibit maturation (or M-phase) promoting factor (MPF) activity, which regulates meiotic progression in oocytes. This paper reviews what is understood of calcium release at fertilization and describes the treatments that have been used to induce oocyte activation artificially in parthenogenetic and nuclear transfer studies. The relative effectiveness of the strategies employed to mimic the oocyte's response to sperm are discussed. Key words: Oocyte activation, Fertilization, Parthenogenesis, Nuclear transfer (J. Reprod. Dev. 48: 2002) o produce a viable embryo at fertilization, the s p e r m n o t o n l y p r o v i d e s t h e m a l e chromosomal complement, but also the stimulus that initiates development. The mechanism by which the activating signal is transmitted from the sperm to the oocyte is not fully clarified, but it is clear that this signal triggers a large increase in the concentration of intracellular-free calcium. In fertilized sea urchin and Xenopus eggs, the calcium increase takes the form of a propagating wave that originates at the point of sperm attachment and traverses the egg at a velocity of 5 to 10 µm/sec

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Section: Calcium Ionophoresmentioning

confidence: 99%

Calcium Release at Fertilization: Artificially Mimicking the Oocyte's Response to Sperm.

Grupen¹,

Nottle²,

Nagashima

2002

Journal of Reproduction and Development

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“…The basic medium (mM199) used for in vitro fertilization was modified Medium 199 [10]. All media were equilibrated at 39 C in an atmosphere of 5% CO2 in air for a minimum of 3 h prior to incubation of oocytes.…”

Section: Culture Mediamentioning

confidence: 99%

The Presence of Tissue Inhibitor of Matrix Metalloproteinase-1 (TIMP-1) During Meiosis Improves Porcine 'Oocyte Competence' as Determined by Early Embryonic Development After In-vitro Fertilization.

Funahashi

McIntush

Smith

et al. 1999

Journal of Reproduction and Development

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Abstract. The present studies were designed to determine the effect of tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) during meiosis or after in vitro fertilization on early embryonic development of porcine embryos. The presence of purified ovine TIMP-1 in culture medium after in vitro fertilization did not affect the incidence of embryo cleavage or development to the blastocyst stage at 48 h or 6 days after insemination, respectively. In contrast, when various concentrations (1.25, 2.50 and 5.00 µg/ml) of purified ovine TIMP-1 were added to the culture medium during the second half (24 h) of oocyte maturation, normal cleavage rates (64 ± 3% at 1.25 µg/ml, 57 ± 5% at 2.50 µg/ml and 56 ± 1% at 5.00 µg/ml) were higher (p<0.05) as compared to controls (43 ± 2%). In addition, the incidence of embryos that developed to the blastocyst stage was increased (p<0.05) when TIMP-1 was added (34 ± 3% at 1.25 µg/ml, 32 ± 1% at 2.50 µg/ml and 28 ± 1% at 5.00 µg/ml) as compared to controls (22 ± 3%). There was no difference in the incidence (53 ± 1%) of oocytes fertilized normally. These results indicate that the presence of TIMP-1 during the second half of in vitro maturation enhanced the competence of porcine oocytes to develop to the 2-to 4-cell stages after in vitro fertilization, consequently increasing the incidence of blastocysts. Key word: Pig, oocyte, TIMP-1, oocyte competence, in vitro maturation (J. Reprod. Dev. 45: [265][266][267][268][269][270][271] 1999) immunological and biological properties similar to those of tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) [1]. TIMP-1 is known to play an important role in the extracellular matrix remodeling of follicular tissue by regulating the activity of matrix metalloproteinases (MMP), which are metal-dependent enzymes capable of degrading proteinaceous components of the extracellular matrix (see review by McIntush and Smith [2]). TIMP-1 is a multifunctional protein that, in addition to inhibiting MMP activity, reportedly

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“…Within 2 h post slaughter, the follicular contents were recovered by cutting from visible small antral follicles (about 2-5 mm in diameter) on the ovarian surface with a razor, and by scraping the inner surface of the follicle walls with a disposable surgical blade. The cumulus-oocyte complexes with uniform ooplasm and a compact cumulus cell mass were prepared in HEPES-buffered Tyrode's medium containing 0.01% (w/v) polyvinyl alcohol (H-TL-PVA, [26]). The maturation medium was BSA-free NCSU37 (NCSU37, [27]) supplemented with 0.6 mM cysteine, 2% (v/v) MEM amino acids solution (Gibco BRL, Grand Island, NY), 1% (v/v) MEM non-essential amino acids solution (Gibco), 0.04 units/ml oFSH (Sigma Chemical Co., St. Louis, MO), 0.02 units/ml oLH (Sigma), and 10% (v/v) pig follicular fluid.…”

Section: Oocyte Maturationmentioning

confidence: 99%

Time Course Analyses of Cortical Granule Exocytosis and Zona Pellucida Modifications after Artificial Activation in In-Vitro Matured Porcine Oocytes.

Tatemoto

Takato

1999

J.Mamm.Ova.Res.

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Abstract:The present study was carried out to determine the time-dependent changes in the cortical granule (CG) distribution and the glycoprotein compositions of zona pellucida (ZP) after egg activation by electrostimulation in porcine oocytes matured in vitro. CG exocytosis was observed by staining with fluorescein isothiocyanate-labeled peanut agglutinin (FITC-PNA) and laser confocal microscopy, and ZP modification was analyzed by using enhanced chemiluminescent (ECL) detection of the biotinylated ZP derived from a single oocyte. In the oocytes matured in vitro, CGs staining with FITC-PNA had formed a continuous monolayer underlying the oolemma, and three major bands (ZP1, ZP2 and ZP3) were observed in a biotinylated ZP subjected to SDS-PAGE followed by ECL detection. Electrostimulation to induce artificial activation caused a decline in the fluorescent intensity of the CGs with a concomitant decrease in the amounts of ZP1 and ZP2 bands, but CG exocytosis according to oocyte activation occurred slowly, and the incidence of oocytes with complete CG exocytosis was first observed 1.0 h after electrostimulation (9%). Similarly, the complete ZP modification, i.e. a maximum decrease in amounts of ZP1 and ZP2 bands, required more than 3.5 h after artificial activation, and the ZP dissolution time caused by 0.1% protease action increased apparently as incubation time was prolonged after electrostimulation. Results of these experiments suggest that the slower ZP modification due to the delayed CG exocytosis in porcine oocytes may reflect the remarkable increase in polyspermy frequency in in-vitro fertilization. Key words: Cortical granule exocytosis, Zona reaction, Zona hardening, ECL detection, Porcine oocyte.All mammalian eggs are surrounded by a relatively thick, insoluble extracellular coat called the zona pellucida (ZP) [1]. After fertilization, the ZP plays an important role in the subsequent activation events [2]. The porcine ZP is composed of three glycoprotein families, ZP1 (92 kDa), ZP3α (55 kDa), and ZP3β (55 kDa), which were named with a nomenclature based on apparent molecular weight [3,4], and ZP1 is split into two smaller molecules, ZP2 (69 kDa) and ZP4 (23 kDa) under reducing conditions [5]. The porcine ZP3 families have been shown to comprise approximately 60% of the total glycoprotein content [4,6]. In particular, ZP3α seems to be responsible for the binding of boar spermatozoa to oocytes, since preincubation of boar sperm with solubilized and purified ZP3α inhibits subsequent sperm attachment to porcine ZP [7].In mammalian species, sperm activates oocytes by triggering calcium oscillations that persist for more than an hour after sperm penetration [8][9][10], and artificially increasing intracellular Ca 2+ by application of electrostimulation in the presence of Ca 2+ [11], or injection of CaCl 2 [12] is capable of inducing parthenogenetic activation in porcine oocytes. The increase in cytoplasmic free-Ca 2+ is recognized as the primary intracellular signal responsible for initiating the activation pr...

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In Vitro Development of in Vitro-Matured Porcine Oocytes Following Chemical Activation or in Vitro Fertilization1

Cited by 112 publications

References 36 publications

Calcium Release at Fertilization: Artificially Mimicking the Oocyte's Response to Sperm.

Calcium Release at Fertilization: Artificially Mimicking the Oocyte's Response to Sperm.

The Presence of Tissue Inhibitor of Matrix Metalloproteinase-1 (TIMP-1) During Meiosis Improves Porcine 'Oocyte Competence' as Determined by Early Embryonic Development After In-vitro Fertilization.

Time Course Analyses of Cortical Granule Exocytosis and Zona Pellucida Modifications after Artificial Activation in In-Vitro Matured Porcine Oocytes.

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