Recovery of Uterine Embryos in Rhesus Monkeys

Hurst, Philip; Jefferies, Kathryn; Eckstein, Peter P.; Wheeler, A. G.

doi:10.1095/biolreprod15.4.429

Cited by 21 publications

(4 citation statements)

References 9 publications

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“…The surgical flushing procedure, performed on cycle days 19-2 1, was a modification of previously described collection techniques (Hurst et al, 1976;Eddy et al, 1975). Following ligation of the cervix with a rubber band, the oviducts were cannulated with polyethylene tubing, and two blunted 18-gauge needles with attached tubing were inserted at the uterine fundus.…”

Section: Methodsmentioning

confidence: 99%

Differentiation of the blastocyst of the rhesus monkey

Enders

Schläfke

1981

Am. J. Anat.

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A method of flushing the oviduct and/or uterus of rhesus monkeys was used to obtain a number of preimplantation stages, of which four cleavage stages and seven blastocysts that were judged to be normal were studied cytologically using transmission electron microscopy. In addition to the usual changes in mitochondria and endoplasmic reticulum that accompany differentiation of the blastomeres, the blastocysts with zonae showed sequestration of areas of cytoplasm. The first indications of junctional complexes were short stretches of parallel membrane with a slightly increased density found in the morula stage. Blastocysts developed typical apical junctional complexes, but in addition showed extensive gap junctions linking trophoblast and inner cell mass, and epiblast and differentiating endoderm. Endodermal differentiation occurred at about the same time that a basal lamina was found under mural trophoblast and epiblast (but not polar trophoblast or endoderm). Enlarged torn zonae were found in association with one blastocyst and unaccompanied by blastocysts, including a case in which the animal subsequently prove to be pregnant. This observation suggests that hatching is a normal feature of zonal escape in this species. The trophoblast of blastocysts without zonae had well-formed apical absorptive areas and, in some instances, long irregular microvilli in the area near the inner cell mass. Cell debris, vacuoles containing debris and isolated cells, although variable, were common features of the preimplantation stage in the rhesus monkey.

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Section: Methodsmentioning

confidence: 99%

Differentiation of the blastocyst of the rhesus monkey

Enders

Schläfke

1981

Am. J. Anat.

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“…The blastocoel follows from swelling and discharge of microlumens One possibility would be to separate one or a few neighboring TE-ICM contacts and expand the blastocoel from this initial gap (14). Alternatively, multiple intercellular gaps could appear between cells, as can be observed between TE cells in electron micrographs of blastocysts from mice (15) and nonhuman primates (16,17), which would merge into a single lumen via unknown mechanisms (3). To investigate this, we used mouse embryos expressing a membrane marker (18) to perform high-resolution imaging at the onset of lumen formation, when blastomeres have completed their fifth cleavage (movie S1).…”

mentioning

confidence: 99%

Hydraulic fracturing and active coarsening position the lumen of the mouse blastocyst

Dumortier

Verge-Serandour

Tortorelli

et al. 2019

Science

191

189

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During mouse pre-implantation development, the formation of the blastocoel, a fluid-filled lumen, breaks the radial symmetry of the blastocyst. The factors that control the formation and positioning of this basolateral lumen remain obscure. We found that accumulation of pressurized fluid fractures cell-cell contacts into hundreds of micrometer-size lumens. These microlumens eventually discharge their volumes into a single dominant lumen, which we model as a process akin to Ostwald ripening, underlying the coarsening of foams. Using chimeric mutant embryos, we tuned the hydraulic fracturing of cell-cell contacts and steered the coarsening of microlumens, allowing us to successfully manipulate the final position of the lumen. We conclude that hydraulic fracturing of cell-cell contacts followed by contractility-directed coarsening of microlumens sets the first axis of symmetry of the mouse embryo.

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“…To our knowledge, there is no signi cant data available in the relevant literature to throw any meaningful light on the issue of sperms from mid-luteal phase coitus entering inside the trophectoderm of blastocysts during the critical period of implantation. Although Enders and Schlafke 12 reported about two blastocysts with fragments of sperms predictably phagocytosed, Hurst et al did not report the presence of any ultrastructural cues of processes related to such phagocytosis 13 . It was unlikely in either study that zona-free blastocysts had any chance to receive fresh ejaculate from coitus, as females were allowed to mate only during the periovulatory period.…”

Section: Main Textmentioning

confidence: 97%

“…These blastocysts were obtained by uterine ushing on days 6-8 after ovulation from female rhesus monkeys who were cohabitating and had regular vaginal intercourse with proven fertile males resulting in natural conceptions as described earlier 11 . It is notable that obtaining zona-free intact blastocysts are rather di cult since they are sticky, and tend to promptly adhere to the uterine epithelium, and hence they are not amenable to retrieval efforts without causing cellular damage 12,13 . This is the rst visual evidence of intact sperm heads having entered the trophectoderm of blastocysts retrieved on days 6-8 after ovulation in the natural conception cycle of female rhesus monkeys.…”

Section: Main Textmentioning

confidence: 99%

Sperm in the implantation-stage blastocyst: Is it a check point of embryo quality?

Huppertz

Sengupta

Kroneis

et al. 2022

Preprint

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The early human embryo derived from fusion of an oocyte with a single sperm undergoes growth and differentiation and results in an implantation-ready blastocyst. To initiate implantation, the blastocyst hatches from the zona pellucida, thus making it accessible to external inputs. Our modelling of sperm distribution through the uterus indicates that 200–5000 sperms following natural coitus during mid-luteal phase are in a position of reaching the implantation-stage blastocyst in the maternal uterus. We indeed have ultrastructural evidence of sperm cells intruding into the trophectoderm cells of uterine zona-free blastocysts obtained from rhesus monkeys. The question arises whether the negotiation between additional sperm and azonal blastocyst is a feature of evolution yielding adaptation processes influencing the developmental fate of an individual embryo or a neutral by-product in placental mammals. This process potentially bears implications in pregnancy success, sperm competition, and human health.

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Recovery of Uterine Embryos in Rhesus Monkeys

Cited by 21 publications

References 9 publications

Differentiation of the blastocyst of the rhesus monkey

Differentiation of the blastocyst of the rhesus monkey

Hydraulic fracturing and active coarsening position the lumen of the mouse blastocyst

Sperm in the implantation-stage blastocyst: Is it a check point of embryo quality?

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