parthenogenetic activation
Dotaz
Zobrazit nápovědu
This study reported facultative parthenogenetic cleavage development of sterlet sturgeon Acipenser ruthenus eggs and quantified the percentage of parthenogenetically developed eggs in relation to the fertilization ability of different females. When eggs were activated in freshwater, 5.1-13.7% of eggs developed parthenogenetically, while among those activated eggs 3.6-9.4% developed to 2 cells, 0.4-4.5% developed to 4 cells, and 0-0.8% developed to 8 cells. The mean percentage of fertilized and parthenogenetically activated eggs among the females was negatively correlated (R(2)=0.71, p=0.036), which indicates that parthenogenetic activation rate of sterlet eggs depends on the quality of eggs in terms of fertilization rate.
- MeSH
- ovum fyziologie MeSH
- partenogeneze fyziologie MeSH
- ryby fyziologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The nitric oxide donor (+)-S-nitroso-N-acetylpenicillamine (SNAP) is capable of inducing parthenogenetic activation in pig oocytes matured in vitro. However, quite a long exposure to the nitric oxide donor, exceeding 10 h, is necessary for successful oocyte activation. Repeated short-term treatment with 2 mm SNAP significantly increased the activation rates despite the fact that the overall exposure time to the nitric oxide donor did not exceed 4 h. With regard to the activation rate, 12 repeated treatments lasting 10 min each were found to be the most efficient regimen (63.3%). The continuous exposure to the nitric oxide donor for the same overall time induced parthenogenetic activation in 12.5% oocytes (2-h continuous treatment with 2 mm SNAP). The development of parthenogenetic embryos increased after repeated short-term treatment with SNAP. After continuous treatment with 2 mm SNAP for 10 h, only 6.7% of the oocytes cleaved, and none developed beyond the 4-cell stage. Thirty-minute treatment repeated four times with 2 mm SNAP induced cleavage in 37.5% of the oocytes, 18.3% developed to the morula stage, and 6.7% reached the blastocyst stage. Based on the results, it is concluded that pulsatile treatment can significantly improve parthenogenetic activation rate when compared with the continuous treatment using nitric oxide donors.
- MeSH
- donory oxidu dusnatého aplikace a dávkování MeSH
- kultivace embrya veterinární MeSH
- kultivované buňky MeSH
- oocyty účinky léků fyziologie MeSH
- partenogeneze účinky léků fyziologie MeSH
- prasata MeSH
- S-nitroso-N-acetylpenicilamin aplikace a dávkování MeSH
- stadium rýhování vajíčka MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- blastocysta cytologie metabolismus účinky léků MeSH
- buněčná diferenciace MeSH
- buněčné dělení MeSH
- finanční podpora výzkumu jako téma MeSH
- krevní proteiny farmakologie metabolismus MeSH
- kultivační média MeSH
- kultivované buňky MeSH
- oocyty cytologie metabolismus účinky léků MeSH
- Sus scrofa MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
For the functional analysis of insect genes as well as for the production of recombinant proteins for biomedical use, clonal transgenic silkworms are very useful. We examined if they could be produced in the parthenogenetic strain that had been maintained for more than 40years as a female line in which embryogenesis is induced with nearly 100% efficiency by a heat shock treatment of unfertilized eggs. All individuals have identical female genotype. Silkworm transgenesis requires injection of the DNA constructs into the non-diapausing eggs at the preblastodermal stage of embryogenesis. Since our parthenogenetic silkworms produce diapausing eggs, diapause programing was eliminated by incubating ovaries of the parthenogenetic strain in standard male larvae. Chorionated eggs were dissected from the implants, activated by the heat shock treatment and injected with the transgene construct. Several transgenic individuals occurred in the daughter generation. Southern blotting analysis of two randomly chosen transgenic lines VTG1 and VTG14 revealed multiple transgene insertions. Insertions found in the parental females were transferred to the next generation without any changes in their sites and copy numbers, suggesting that transgenic silkworms can be maintained as clonal strains with homozygous transgenes. Cryopreservation was developed for the storage of precious genotypes. As shown for the VTG1 and VTG14 lines, larval ovaries can be stored in DMSO at the temperature of liquid nitrogen, transferred to Grace's medium during defrosting, and then implanted into larvae of either sex of the standard silkworm strains C146 and w1-pnd. Chorionated eggs, which developed in the implants, were dissected and activated by the heat shock to obtain females (nearly 100% efficiency) or by a cold shock to induce development to both sexes in 4% of the eggs. It was then possible to establish bisexual lines homozygous for the transgene.
- MeSH
- bourec genetika MeSH
- embryonální vývoj MeSH
- geneticky modifikovaná zvířata MeSH
- hmyzí geny * MeSH
- kryoprezervace metody MeSH
- partenogeneze MeSH
- reakce na tepelný šok MeSH
- technika přenosu genů MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- blastomery metabolismus ultrastruktura MeSH
- elektrická stimulace MeSH
- finanční podpora výzkumu jako téma MeSH
- intracytoplazmatické injekce spermie MeSH
- oocyty fyziologie MeSH
- partenogeneze MeSH
- skot MeSH
- vakuoly ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
Nitric oxide (NO) is identified as a signaling molecule involved in many cellular or physiological functions including meiotic maturation and parthenogenetic activation of mammalian oocytes. We observed that nitric oxide donor SNAP was potent to induce parthenogenetic activation in Xenopus eggs. NO-scavenger CPTIO impaired the effects of SNAP, providing evidence for the effects of the latter to be specific upon NO release. In Xenopus eggs, SNAP treatment induced pigment rearrangement, pronucleus formation and exocytosis of cortical granules. At a biochemical level, SNAP exposure lead to MAPK and Rsk inactivation within 30 minutes whereas MPF remained active, in contrast to calcium ionophore control where MPF activity dropped rapidly. MAPK inactivation could be correlated to pronuclear envelope reformation observed. In SNAP-treated eggs, a strong increase in intracellular calcium level was observed. NO effects were impaired in calcium-free or calcium limited medium, suggesting that that parthenogenetic activation of Xenopus oocytes with a NO donor was mainly calcium-dependent.
- MeSH
- aktivace enzymů účinky léků MeSH
- aparát dělícího vřeténka účinky léků metabolismus MeSH
- benzoáty farmakologie MeSH
- donory oxidu dusnatého farmakologie MeSH
- faktor podporující zrání metabolismus MeSH
- imidazoly farmakologie MeSH
- kinetika MeSH
- mitogenem aktivované proteinkinasy metabolismus MeSH
- morfogeneze účinky léků MeSH
- ovum cytologie účinky léků metabolismus MeSH
- oxid dusnatý metabolismus MeSH
- partenogeneze MeSH
- progesteron farmakologie MeSH
- S-nitroso-N-acetylpenicilamin farmakokinetika MeSH
- vápník metabolismus MeSH
- Xenopus laevis metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
