Thiocyanate Assay for Sublytic Change in the Zona Pellucida of the Mouse Egg1

Domon, M.; Pinsker, M. Craig; Mintz, Beatrice

doi:10.1093/biolreprod/9.3.246

Cited by 13 publications

(7 citation statements)

References 11 publications

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“…Plasminogen is a constituent of the uterine environment, and serves as a readily utilizable source of protease for the early embryo (Fazleabas et ai, 1983). Plasminogen activation to plasmin may be a mechanism which the embryo uses to effect a 'sublysis' (Domon, Pinsker & Mintz, 1973) of the zona pellucida to facilitate hatching. Menino & Wright (1982) have observed a decrease in the resistance of the pig zona pellucida to pronase with advancing cell stage.…”

Section: Methodsmentioning

confidence: 99%

Enhancement of hatching and trophoblastic outgrowth by mouse embryos cultured in Whitten's medium containing plasmin and plasminogen

Menino¹,

O'Claray²

1986

Reproduction

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Summary. Mice were induced to superovulate and 2-cell embryos were cultured in Whitten's medium with 10 mg bovine serum albumin/ml (WM) as control, Medium WM with 2\m=.\3,4\m=.\6,23\m=.\1 or 46\m=.\2\ g=m\ gplasmin/ml, Medium WM with 14\m=.\6,29\m=.\1 or 145\m=.\7 \ g = m \ g plasminogen/ml, Medium WM with 0\m=.\1,0\m=.\2, 1\m=.\1or 2\m=.\2 \g=m\gtrypsin/ml, Medium WM with 0\m=.\2,0\m=.\3, 1\m=.\6or 3\m=.\3 \g=m\gpronase/ml and Medium WM with 10% heat-treated bovine serum (HTBS). Proteolytic activities in the culture media were evaluated at the start of the culture period and 10 days later. Blastocyst formation was significantly reduced in cultures supplemented with pronase and in the two higher levels of trypsin when compared to that in Medium WM. More embryos developed to the blastocyst stage in Medium WM + 2\m=.\3 or 23\m=.\1\ g=m\ gplasmin/ml and Medium WM + 14\m=.\6\ g=m\ gplasminogen/ ml than in Medium WM (P < 0\m=.\05). The incidence of hatching was significantly greater in Medium WM than in all plasminogen-and plasmin-supplemented media except for Medium WM + 29\m=.\1 \g=m\gplasminogen/ml. Although not significantly different, hatching was lower in Medium WM and Medium WM + 0\m=.\1 \g=m\gtrypsin/ml when compared to Medium WM + HTBS. Similar numbers of embryos completed the hatching process in Media WM, WM + 0\m=.\1 or 0\ m=. \ 2\ g=m\ gtrypsin/ml and WM + 0\m=.\3 \ g = m \ g pronase/ml. Since dissolution of the zona pellucida occurred within 96 h for embryos cultured in Media WM + 1\ m=. \ 6 or 3\ m=. \ 3\ g=m\gpronase/ml and WM + 1\ m=. \ 1 or 2\ m=. \ 2\ g=m\gtrypsin/ ml, hatching could not be evaluated. The incidence of attachment and subsequent trophoblastic outgrowth was significantly greater in media with plasmin, plasminogen and HTBS than in pronase-and trypsin-supplemented media or in Medium WM alone. The results suggest that the enhancement in embryo development observed in pl asmi n\x=req-\ and plasminogen-supplemented media is due not only to the provision of protease but to an additional trophic effect as well. Simple medium supplemented with plasmin or plasminogen is as able to support in-vitro development as is medium supplemented with serum.

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Section: Methodsmentioning

confidence: 99%

Enhancement of hatching and trophoblastic outgrowth by mouse embryos cultured in Whitten's medium containing plasmin and plasminogen

Menino¹,

O'Claray²

1986

Reproduction

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“…The droplet of uterine fluid was covered with paraffin oil and allowed to incubate with zonae for 24 h at 37°C in a humidified incubator. The embryos and zonae were recovered and placed in a 1-3 M-sodium thiocyanate (NaSCN) solution containing 0-25% gelatin and 0-5 M-NaHP04 (pH 7-4, 25°C; Domon et al, 1973). They were examined with a compound microscope at 5-10-min intervals (more often as dissolution approached), and the time required for complete dissolution was determined.…”

Section: Methodsmentioning

confidence: 99%

“…In addition, it has been proposed that the factor, possibly a proteolytic enzyme or enzyme complex (Bowman & McLaren, 1970;Mintz, 1972;Pinsker, Sacco & Mintz, 1974), is also responsible for sublytic changes in the zona before outright lysis (Domon, Pinsker & Mintz, 1973) and altering the surface of the blastocyst (Mintz, 1972;. Although the lytic factor has not been isolated, studies on the dissolution of zonae in vivo indicate that maximal amounts of zona-lytic activity are present approximately 18 h after the injection of oestradiol into mice treated with progesterone and that lysis of zonae did not occur in animals treated with either hormone alone (Hoversland & Weitlauf, 1981b).…”

Section: Introductionmentioning

confidence: 99%

In-vitro zona-lytic activity in uterine fluid from ovariectomized mice treated with oestradiol-17 and progesterone

Hoversland¹,

Weitlauf²

1982

Reproduction

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Mouse embryos collected before implantation were incubated in vitro for 24 h with fluid rinsed from the uteri of ovariectomized female mice injected with progesterone, oestradiol-17 beta + progesterone, oestradiol-17 beta + progesterone, or oestradiol-17 beta alone. Although none of the zonae was completely dissolved, those incubated in fluid from animals treated with oestradiol + progesterone were subsequently more soluble in sodium thiocyanate (NaSCN) than those incubated similarly in control buffer, indicating a sublytic change during the incubation with uterine washings. Zonae incubated in fluid from animals injected with either hormone alone did not undergo such a change.

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“…Deficiency diseases of this enzyme are characterized by an accumulation of glycopotein and GMp Enders and Schlafke (1974) detected a thin mucopolysaccharide coat on the embryo cell surface. Pinsker and Mintz (1973) found changes in surface carbohydrate content of late stage embroys. "Lets" glycoprotein appears on the surface of the endoderm cells in the inner cell mass (Zetter and Martin, 1978;Wartiovaara, Leivo and Vaheri, 1979).…”

Section: -Galactosidase Activity In Preimplantation Mouse Embryosmentioning

confidence: 97%

The expression of the [Bgl] genetic complex during preimplantation mouse embryogenesis and in ovulated eggs

Esworthy

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Thiocyanate Assay for Sublytic Change in the Zona Pellucida of the Mouse Egg1

Cited by 13 publications

References 11 publications

Enhancement of hatching and trophoblastic outgrowth by mouse embryos cultured in Whitten's medium containing plasmin and plasminogen

Enhancement of hatching and trophoblastic outgrowth by mouse embryos cultured in Whitten's medium containing plasmin and plasminogen

In-vitro zona-lytic activity in uterine fluid from ovariectomized mice treated with oestradiol-17 and progesterone

The expression of the [Bgl] genetic complex during preimplantation mouse embryogenesis and in ovulated eggs

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