Effects of Ions on the Development of the Pre-Implantation Mouse Embryo in Vitro

Rg, Wales

doi:10.1071/bi9700421

Cited by 56 publications

(15 citation statements)

References 8 publications

(12 reference statements)

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“…Our results corroborate the findings of Bilozur & Powers (1982). However, while extracellular calcium appears to be necessary for the induction and continuous maintenance of compaction (Wales, 1970;Whitten, 1971;Ducibella & Anderson, 1975 ;Bilozur & Powers, 1982) (Schultz, Heller & Buklad, 1983). Calmodulin has been implicated in oocyte maturation in non-mammalian species (Meijer & Guerrier, 1981;Wasserman & Smith, 1981;Hollinger & Alvarez, 1982).…”

Section: Resultssupporting

confidence: 82%

Role of calmodulin in blastocyst formation in the mouse

Pakrasi¹,

Dey²

1984

Reproduction

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Summary. Preimplantation mouse embryos were recovered by flushing the oviducts on Day 3 at 09:30\p=n-\10:00h, 15:30\p=n-\16:00h and 21:30\p=n-\22:00h: When placed in culture for 48 h, 79% of the 4\p=n-\8cell embryos recovered at 09:30\p=n-\10:00h developed into blastocysts, but a large number of these embryos failed to form blastocysts when exposed to trifluoperazine, a calmodulin antagonist, at 0\m=.\5or 0\m=.\6 \g=m\m in culture. About 45% of the embryos recovered at 15:30\p=n-\16:00h were compacted and blastocyst formation was again markedly depressed in the presence of the drug. Advanced compacted embryos recovered at 21:30\p=n-\22:00 h showed normal development into blastocysts in the presence of 0\m=.\6\g=m\m-trifluoperazine. Trifluoperazine sulphoxide (the inactive form of trifluoperazine) at 0\m=.\6or 1\m=.\2 \g=m\m concentration had no effect on blastocyst formation of uncompacted embryos recovered at 09:30\p=n-\10:00h. These embryos and those recovered at 21:30\p=n-\22:00 h and developed into blastocysts in the presence of trifluoperazine were transferred to Day-4 pseudopregnant mice and healthy young were born. When exposed to calcium-free medium or medium containing trifluoperazine all compacted embryos recovered at 18:30 h became decompacted; development to the blastocyst stage was normal in medium alone but reduced when trifluoperazine was added. Compacted embryos recovered at 23:00 h showed 100% decompaction in the calcium-free medium but completely failed to decompact in the presence of 0\m=.\6 \g=m\m-trifluoperazine. We suggest that extracellular calcium is essential for the continuance of compaction, while intracellular calcium is required only during the initial phase of this process.

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Section: Resultssupporting

confidence: 82%

Role of calmodulin in blastocyst formation in the mouse

Pakrasi¹,

Dey²

1984

Reproduction

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“…However, this medium probably does not simulate physiological conditions very precisely. Calcium ions are required for the in vitro development of mouse embryos (Wales 1970) and bicarbonate is superior to phosphatebuffered medium in maintaining growth and metabolism of cultured mouse embryos (Quinn and Wales 1973). Thus the use of Krebs-Ringer bicarbonate medium, which contains similar concentrations of calcium and bicarbonate to those found in rabbit fallopian tube fluid (David et al 1969), would probably give results with more physiological significance than those obtained with calcium-free Krebs-Ringer phosphate medium.…”

mentioning

confidence: 96%

The in Vitro Metabolism of [U-14C]Glucose by the Preimplantation Rabbit Embryo

Quinn¹,

Wales²

1973

Aust. Jnl. Of Bio. Sci.

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The incorporation of glucose carbon into various components of embryos and culture medium was studied throughout the preimplantation period of development in the rabbit embryo.During the first 3 days after ovulation, embryos were able to develop for 24 hr in a simple chemically defined medium containing glucose and serum albumin. After blastocyst formation, this medium could maintain normal morphological appearance of day 4, day 5, and day 6 blastocysts for at least 3-4 hr.The uptake of glucose carbon by the embryo increased as development progressed, with substantial increases in the incorporation of glucose occurring after blastocyst formation. A considerable amount of the label incorporated was present in the embryos as the parent substrate, indicating that the uptake of glucose was greater than its conversion to other metabolites. Nevertheless, throughout development glucose carbon was incorporated into a wide variety of the carbon pools of the embryo, such as acidic and basic acid-soluble compounds, proteins, and lipids. Lactate, pyruvate, and acetate also accumulated in the culture medium. The rate of protein synthesis declined in relation to the increase in cell number during development and may reflect the requirements of the rabbit embryo for exogenous protein and amino acids for blastocyst expansion.A considerable amount of the incorporation of substrate carbon into day 6 blastocysts was due to the accumulation of lactate, pyruvate, acetate, amino acids, and the parent substrate within the blastocoel fluid. The higher concentration of metabolites, such as carboxylic acids and amino acids, in the blastocoel fluid than in the incubation medium indicated that there was some active mechanism for retaining these compounds within the blastocyst. The blastocyst tissue also contained labelled metabolites such as lactate, citrate, and malate as well as the amino acids glutamate, alanine, and aspartate. The accumulation of glucose carbon in these metabolites indicates that glucose is metabolized in the rabbit embryo by the operation of the Embden-Meyerhof pathway and tricarboxylic acid cycle.

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“…7-5 ± 0-03 7-5 ± 0-00 Wales (1970), who determined that embryo development occurs within a wide range of phosphate concentrations and that only by removing phosphate completely will embryo development be reduced significantly. However, Schini & Bavister (1988) have recently reported that phosphate at any concentration is detrimental to hamster embryo development.…”

Section: Tested Mediamentioning

confidence: 99%

Optimization of mouse embryo culture media using simplex methods

Lawitts

Biggers²

1991

Reproduction

216

112

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Summary. Culture media were developed for pronuclear-stage mouse embryos using simplex optimization, which has the benefit of being able to optimize several components simultaneously. Initially, several different media were generated. All Another experiment compared 4 of the best media generated from the simplex optimization. In 3 out of the 4 media, 90% or more of the embryos were \m=ge\4cells on Day 3. In 3 of the media, approximately 60% or more of the embryos developed into blastocysts. The simplex optimization procedure is an efficient method for developing culture media and determining requirements for development in vitro.

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Effects of Ions on the Development of the Pre-Implantation Mouse Embryo in Vitro

Cited by 56 publications

References 8 publications

Role of calmodulin in blastocyst formation in the mouse

Role of calmodulin in blastocyst formation in the mouse

The in Vitro Metabolism of [U-14C]Glucose by the Preimplantation Rabbit Embryo

Optimization of mouse embryo culture media using simplex methods

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