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[Parthenogenesis].

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Parthenogenesis↗

Cytoskeletal alteration in aged porcine oocytes and parthenogenesis.

The objective of this study was to examine the changes in microtubule and microfilament assembly in aged porcine oocytes and to determine their developmental pattern after parthenogenetic activation. Porcine oocytes were cultured in Whitten's medium containing 10% follicular fluid with hormonal supplements (eCG and hCG) for 22 hr and 40 hr additional culture without hormonal supplements. At 40, 50, and 60 hr of culture, the oocytes were fixed for immunocytochemistry or activated by electrical pulse. In metaphase II stage oocytes, microtubules were detected only in the meiotic spindle. Two microfilament domains existed in the egg cortex, a thick and a thin microfilament domain. In aged oocytes (50 and 60 hr of culture), the incidence of metaphase II plates observed outside of the thick microfilament domain was higher (P < 0.05) than in young oocytes (40 hr of culture). After activation, a polar body was usually emitted from the chromatin at the microfilament rich domain or two pronuclei were formed outside of the microfilament rich domain. The percentage of activated oocytes with one female pronucleus was higher (P < 0.05) in oocytes at 40 hr of maturation than at 50 and 60 hr of culture. At 24 and 30 hr after stimulation the incidence of cleavage to the 3- to 4-cell stage was higher (P < 0.05) in aged oocytes (50 and 60 hr of maturation) than that in oocytes at 40 hr of culture. These results suggested that a role of microfilaments is to retain the chromatin at the proper position in the oocyte cortex, and that aging results in a disruption of the microfilaments such that atypical development results after parthenogenetic activation.

Actin Cytoskeleton↗

Bovine parthenogenesis is characterized by abnormal chromosomal complements: implications for maternal and paternal co-dependence during early bovine development.

The present study was conducted to examine the karyotypes of parthenogenetic bovine embryos arising from the application of standard oocyte activation and diploidization methods. Bovine cumulusoocyte complexes were collected and matured in vitro for 24 hr prior to oocyte activation with either 5 microM ionomycin or 7% ethanol for 5 min. Groups of activated oocytes were further treated with 5 micrograms/ml cytochalasin D or 1.9 mM 6-dimethylaminopurine (DMAP) for 6 hr. Cleavage varied significantly (P < .05) among the treatment groups with 68.0% of the ethanol- and DMAP-treated oocytes dividing. Blastocyst development did not vary with 18.4 +/- 2.5% of all treated oocytes progressing to this stage. Blastocyst development did not occur in groups subjected to oocyte activation alone. Blastocysts displayed haploid (2.3%), diploid (11.4%), tetraploid (40.9%), octaploid (4.5%), and mixoploid chromosomal complements (40.9%). Two-cell stage parthenogenotes resulting from ethanol or ionomycin treatment alone displayed haploid (66.7%), diploid (16.7%), tetraploid (4.2%), and mixoploid (12.5%) complements. Our results demonstrate that diploid bovine parthenogenotes arising from these procedures are a minority, with the majority of parthenogenotes displaying polyploid and mixoploid chromosomal complements. The events contributing to these abnormal chromosomal complements occur as early as completion of the first cell cycle, possibly linking these events with the absence of a paternally supplied centrosome.

Adenine↗

Cytogenetic analysis of ethanol-induced parthenogenesis.

The brief exposure of recently ovulated mouse oocytes to a dilute solution of ethanol in vitro for 1, 3, or 5 min induced a uniform high incidence of parthenogenetic activation. The majority of parthenogenones developed a single haploid pronucleus after the extrusion of a second polar body. The proportionate incidence of this parthenogenetic class was significantly reduced as the duration of ethanol exposure increased from 1 min to 5 min. There was a concomitant increase in the incidence of parthenogenones that developed two haploid pronuclei following failure of extrusion of the second polar body. Cytogenetic analysis of the ethanol-induced single-pronuclear haploid parthenogenones at metaphase of the first cleavage division clearly demonstrated that a significant proportion were aneuploid. The incidence of aneuploidy observed was directly related to the duration of ethanol exposure. G-band analysis of the aneuploid metaphases revealed that the chromosomes were not randomly involved in the malsegregation events. This observation may be a reflection of the relationship of particular chromosomes to the meiotic spindle apparatus rather than on any specific property of the agent to which they were exposed. It is believed that ethanol disrupts the organisation of cytoskeletal elements and, in particular, interferes with the processes of chromosome segregation at the second meiotic division.

Aneuploidy↗

Nuclear dynamics of parthenogenesis of bovine oocytes matured in vitro for 20 and 40 hours and activated with combined ethanol and cycloheximide treatment.

Bovine oocytes matured in vitro (IVM) for 20 hr vs. 40 hr were treated for activation with 7% ethanol in Dulbecco's phosphate-buffered saline for 5 min followed by incubation in M199 + 7.5% fetal calf serum containing cycloheximide (10 micrograms/ml). Treated IVM oocytes and the controls (no ethanol and cycloheximide exposures) were fixed after 0, 1, 2, 3, 4, 5, 7, 10, and 20 hr of incubation and stained 24 hr later with 1% acetoorcein to examine nuclear events. Different stages of nuclear development of the activated oocytes were identified on the basis of nuclear and chromosomal morphology. Pronuclear development was classified into four stages (PN I, II, III, and IV) according to pronuclear progression in chromatin decondensation, nucleoplasm appearance, and nuclear size. The results demonstrated that the combined activation treatment effectively drove the IVM oocytes, both young (20 hr) and aging (40 hr), out of metaphase arrest. The activation rates for young oocytes examined immediately after 0, 1, 2, 3, 4, 5, 7, 10, and 20 hr of incubation with cycloheximide were, respectively, 7%, 24%, 77%, 96%, 92%, 97%, 98%, 93%, and 98%. For aging oocytes (40 hr) the corresponding activation values at the same time intervals were 6%, 84%, 100%, 100%, 100%, 100%, 98%, 100%, and 100%, respectively. These values were significantly higher than those for the corresponding controls. The activated aging oocytes achieved peak activation response more rapidly than did young oocytes. In addition, nuclear events in aging oocytes proceeded faster than those in young ones. Spontaneous activation rates of the aging oocytes were also higher (6-57%) than those of the young ones (0-14%).

Anaphase↗

Patterns in plant parthenogenesis.

Plant taxa that reproduce asexually display some distinct geographical and ecological patterns. A literature review reveals that such taxa 1) tend to have larger ranges, 2) tend to range into higher latitudes, and 3) tend to range to higher elevations than do their sexual relatives. Asexual taxa have a greater tendency than sexual taxa do to colonize once-glaciated areas. These trends have previously been identified as characteristic of parthenogenetic animals as well. While many authors have interpreted these trends as providing support for the 'biotic uncertainty' hypothesis for the maintenance of sex, these trends are consistent with several other interpretations as well. Furthermore, all of these interpretations have ignored the positive correlation that exists between ploidy level and breeding system: asexual plant and animal taxa are generally polyploid, while their sexual relatives are generally diploid. Evidence is presented for plants, and by extension for animals as well, that high ploidy levels alone-independent of breeding system-could endow individuals with the ability to tolerate these 'extreme' environments. For this reason, it appears premature to interpret observed distribution patterns as evidence to support hypotheses about what forces maintain sexual reproduction. Only experimental tests, using sexuals and asexuals of comparable ploidy levels, can permit us to discriminate among the alternatives.

Ecology↗

Spontaneous parthenogenesis in Mus musculus: comparison of protein synthesis in parthenogenetic and normal preimplantation embryos.

In preimplantation stages of normal and spontaneously activated parthenogenetic embryos of the LT/Sv mouse strain, protein synthesis was analyzed by using two-dimensional polyacrylamide gel electrophoresis. Fertilization and parthenogenetic activation cause similar changes polypeptide synthesis when compared with those of unfertilized eggs. The overt developmental delay of early parthenotes, which is probably due to an initial retarded activation in comparison with normal fertilization, is documented molecularly by a similar delay in their protein synthesis pattern. These differences are clearly visible at the two-cell stage but gradually disappear during further cleavage. The basic protein patterns of normal and parthenogenetic embryos are remarkably similar up to the blastocyst stage. However, quantitative differences occur in all preimplantation embryos analyzed and become more distinct at the blastocyst stage. In addition, only minor qualitative changes appear during late preimplantation. These alterations in protein synthesis may reflect at the molecular level early events in abnormal development of parthenotes. Our biochemical results are discussed in context with biological experiments rescuing parthenogenetic LT/Sv embryos by chimera formation.

Animals↗