Partial Characterization of the Plasminogen Activator Produced by Ovine Embryos in Vitro1

Bartlett, Samuel L.; Menino, Alfred R.

doi:10.1095/biolreprod49.2.381

Cited by 9 publications

(5 citation statements)

References 24 publications

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“…Although cryopreservation has been shown to affect embryo metabolism we have been unable to find in the literature any information on the protein secretions of vitrified/thawed embryos, after a short time of in vitro culture. The main group of proteins found in blastocyst secretion after thawing has a MW of 48-50 kDa and proteolytic activity which after zymographic analysis can be identified as urokinase type PA. PA is secreted during blastocoelic expansion and blastocyst hatching (Berg and Menino, 1992;Bartlett and Menino, 1993), participates in cell migration and tissue remodelling during embryogenesis (Fazleabas et al, 1983) and can be involved in the weakening of the zona pellucida and hatching process (Menino et al, 1989;Leoni et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

Resumption of metabolic activity of vitrified/warmed ovine embryos

Leoni

Berlinguer

Rosati

et al. 2002

Molecular Reproduction Devel

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To evaluate resumption of metabolic activity of vitrified ovine embryos during a short time immediately after warming, blastocysts collected from superovulated Sarda ewes were incubated with (35)S-methionine. In vitrified/warmed embryo groups the protein secretion significantly (P < 0.05) increased from 0 to 24 hr of culture, reaching significantly (P < 0.01) higher activity at 18-24 hr and dropping to values similar to the control nonvitrified embryo group at 29-35 hr. Within the control group at 29-35 hr there was a significantly (P < 0.01) higher level of protein secretion compared to the other interval times. The electrophoretic pattern showed a 48-50 kDa secreted protein identified as urokinase-type plasminogen activator (PA). The caseinolytic assay of PA activity showed a course similar to protein secretion in both vitrified and control groups. During 29-35 hr of culture, we did not observe any improvement in PA activity as seen for secreted proteins. At this time, we observed the secretion of a new 20 kDa protein that was not present in vitrified/warmed embryos. Analysis of BrDU incorporation in newly synthesised DNA showed a significant (P < 0.01) improvement in positive cell number from 3 to 9 hr after warming, reaching a value similar to that of the control group at 12 hr of culture. Our results suggest that vitrified/warmed embryos require 9-12 hr of culture to complete resumption of DNA synthesis and 29-35 hr to re-acquire the full capacity of protein secretion but not the qualitative secretion pattern.

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Section: Discussionmentioning

confidence: 99%

Resumption of metabolic activity of vitrified/warmed ovine embryos

Leoni

Berlinguer

Rosati

et al. 2002

Molecular Reproduction Devel

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“…Likewise, cathepsin-L proteolytic activity and protein has been detected in endometrial GE and uterine flushings from early implanting pigs . Urokinasetype plasminogen activator (uPA) is produced by mouse [Strickland et al, 1976], pig [Fazleabas et al, 1983] and ovine [Bartlett and Menino, 1993] embryos, and has been postulated to play a role in implantation of blastocysts into the rodent uterus [Sappino et al, 1989]. Further, the uPA receptor interacts with uPA and integrins to both facilitate a proteolytic cascade at the cell surface, and initiate signaling events that contribute to cell adhesive processes in a non-proteolytic manner [Ossowski and Aguirre-Ghiso, 2000;Preissner et al, 2000].…”

Section: Potential Integrin/matrix-activating Components Of Histotrophmentioning

confidence: 99%

Integrins and Extracellular Matrix Proteins at the Maternal-Fetal Interface in Domestic Animals

Burghardt¹,

Johnson

Jaeger³

et al. 2002

Cells Tissues Organs

157

133

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Establishment of pregnancy in mammals requires coordinated conceptus-maternal interactions involving numerous hormones, growth factors and cytokines acting via specific receptors in the uterus. Uterine secretions play an important role in establishing synchrony between development of the conceptus and uterine receptivity, as well as in conceptus remodeling, adhesion, implantation and placentation in domestic species. Studies of non-invasive implantation in domestic livestock provide valuable opportunities to investigate fundamental processes of the initial events of apposition, attachment and adhesive interactions that are shared among species. In pigs and sheep, it appears that integrins play a dominant role in these fundamental processes via interactions with extracellular matrix molecules and other ligands to transduce cellular signals in uterine epithelial cells and conceptus trophectoderm. This review considers several of the potential integrin-binding ligands involved in the complex implantation adhesion cascade in pigs and sheep along with in vitro evidence for the transduction of cytoplasmic signals that may be required to sustain fetal and maternal contributions to the formation of the epitheliochorial placenta.

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“…Although PA activity in rat [3], mouse [4,5], porcine [6], ovine [7,8], and bovine [9][10][11][12] embryos, and trypsinlike activity in hamster [13] and mouse [14] oocytes have been reported, reports describing proteases in the mammalian oocyte itself are limited. The first identification of an oocyte-produced protease was reported by Huarte et al [15], who found that resumption of meiosis in mouse and rat oocytes triggers the production of tPA.…”

Section: Introductionmentioning

confidence: 99%

Production of Plasminogen Activators (PAs) in Bovine Cumulus-Oocyte Complexes during Maturation In Vitro: Effects of Epidermal Growth Factor on Production of PAs in Oocytes and Cumulus Cells1

Park

Choi

Song

et al. 1999

Biology of Reproduction

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We examined whether plasminogen activators (PAs) are produced by bovine cumulus-oocyte complexes (COCs) during maturation in vitro. The effects of epidermal growth factor (EGF) on production of PAs in oocytes and cumulus cells were also examined. When COCs were cultured for 24 h with 30 ng/ml EGF, three plasminogen-dependent lytic zones (58.5 +/- 3.5 kDa, 79.0 +/- 3.0 kDa, and 113.5 +/- 6.5 kDa) were observed. Addition of amiloride, a competitive inhibitor of urokinase-type PA (uPA), to the zymogram eliminated the activity of the 58.5 +/- 3.5-kDa zone, suggesting that this band is a uPA. However, since the activity of the remaining two bands was not eliminated, it was suggested that the 79.0 +/- 3.0-kDa band is a tissue-type PA (tPA) and the 113.5 +/- 6.5-kDa band is possibly a tPA-PA inhibitor (tPA-PAI) complex. In COCs before culture, however, no activity of PAs was detected. At 6 h of culture, the same level of uPA activity was detected in COCs cultured both in the absence and in the presence of EGF. The uPA activity was increased at 12 h of culture but without further increase at 24 h of culture, with higher activity in the presence than in the absence of EGF. The activity of tPA and tPA-PAI was first detected at 24 h of culture in the absence of EGF. In the presence of EGF, however, some activity of tPA-PAI was detected at 12 h of culture. At 24 h of culture, the activity of all PAs was detected in cumulus cells, but only uPA activity was detected in oocytes, with higher activity in the presence than in the absence of EGF. The uPA activity in oocytes was not detected when they were cultured without cumulus cells in either the presence or absence of EGF, although cumulus expansion was stimulated by EGF, exhibiting a time-course similar to that observed in PA production. These results suggest that uPA, tPA, and tPA-PAI are all produced by bovine COCs, but only uPA by oocytes, during maturation in vitro. However, cumulus cells play an essential role or roles in the production of uPA by oocytes, and EGF enhances the roles of cumulus cells.

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Partial Characterization of the Plasminogen Activator Produced by Ovine Embryos in Vitro1

Cited by 9 publications

References 24 publications

Resumption of metabolic activity of vitrified/warmed ovine embryos

Resumption of metabolic activity of vitrified/warmed ovine embryos

Integrins and Extracellular Matrix Proteins at the Maternal-Fetal Interface in Domestic Animals

Production of Plasminogen Activators (PAs) in Bovine Cumulus-Oocyte Complexes during Maturation In Vitro: Effects of Epidermal Growth Factor on Production of PAs in Oocytes and Cumulus Cells1

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