Chapter 5 Production of Haploid and Diploid Androgenetic Zebrafish (Including Methodology for Delayed in Vitro Fertilization)

Corley-Smith, Graham E.; Brandhorst, Bruce P.; Walker, Charline; Postlethwait, John H.

doi:10.1016/s0091-679x(08)61820-8

Cited by 17 publications

(11 citation statements)

References 18 publications

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“…As previously described, 32 an egg's capacity to be fertilized is compromised when aged in vitro; the longer the period between collection and fertilization, the lower the developmental rate ( Fig. 3).…”

Section: Mpf Activity Cyclin B and Phosphorylated Cdc2 In Unfertilisupporting

confidence: 52%

“…30 Two major holding media are known to maintain the nonactivated stage of zebrafish eggs: Hank's balanced salt solution with 0.5% bovine serum albumin (H-BSA; osmolarity 290 mosmol=L) and Chinook salmon ovarian fluid (CSOF; osmolarity 298 AE 6 mosmol=L). H-BSA is reported to maintain the fertilization capacity of eggs for up to 1 h, 31 whereas CSOF has done so for up to 6 h. 32 An experiment was conducted to reevaluate the capacity of CSOF to maintain eggs in vitro and the subsequent 98 SIRIPATTARAPRAVAT ET AL.…”

Section: Mpf Activity Cyclin B and Phosphorylated Cdc2 In Unfertilimentioning

confidence: 99%

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Characterization andIn VitroControl of MPF Activity in Zebrafish Eggs

et al. 2009

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We describe the characterization of maturation-promoting factor (MPF) in zebrafish eggs and used different defined conditions to maintain its activity in vitro. MPF activity levels are high in freshly ovulated mature eggs and decline rapidly within 5 min after either fertilization or parthenogenetic activation. The MPF activity of eggs matured in vitro declines faster when the eggs are incubated in Hank's culture medium supplemented with 0.5% BSA (H-BSA) than when incubated in Chinook salmon ovarian fluid (CSOF). MPF activity in nonactivated, aged eggs remains high in H-BSA supplemented with 75 microM MG132 or 10 mM caffeine, but neither MG132 nor caffeine can sustain high MPF activity in activated eggs. MG132-treated eggs showed delayed completion of metaphase and extrusion of the second polar body. Nuclear staining of the activated eggs confirmed the correlation between their cell cycle stage and MPF activity at each time point. An embryotoxic effect was found when matured eggs were held in 100 microM of MG132 or 20 mM caffeine for 1 h. Calcium-depleted medium and 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid also showed detrimental effects on the embryos. Conversely, nonactivated, aged matured eggs maintained high MPF activity and developmental potential when CSOF was used as a holding medium.

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Section: Mpf Activity Cyclin B and Phosphorylated Cdc2 In Unfertilisupporting

confidence: 52%

Section: Mpf Activity Cyclin B and Phosphorylated Cdc2 In Unfertilimentioning

confidence: 99%

Characterization andIn VitroControl of MPF Activity in Zebrafish Eggs

et al. 2009

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“…At Ϸ4-6 months of age, eggs were expressed from anesthetized F 1 females and fertilized in vitro with UV-inactivated sperm to generate haploid embryos as described in ref. 18. Thirty-six-hpf (hours postfertilization) haploid embryos were used for whole-mount immunohistochemistry with anti-phosphohistone H3 (pH3).…”

Section: Methodsmentioning

confidence: 99%

A zebrafish bmyb mutation causes genome instability and increased cancer susceptibility

Shepard

Amatruda

Stern

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

154

157

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A major goal of cancer research has been to identify genes that contribute to cancer formation. The similar pathology between zebrafish and human tumors, as well as the past success of large-scale genetic screens in uncovering human disease genes, makes zebrafish an ideal system in which to find such new genes. Here, we show that a zebrafish forward genetic screen uncovered multiple cell proliferation mutants including one mutant, crash&burn (crb), that represents a loss-of-function mutation in bmyb, a transcriptional regulator and member of a putative protooncogene family. crb mutant embryos have defects in mitotic progression and spindle formation, and exhibit genome instability. Regulation of cyclin B levels by bmyb appears to be the mechanism of mitotic accumulation in crb. Carcinogenesis studies reveal increased cancer susceptibility in adult crb heterozygotes. Geneexpression signatures associated with loss of bmyb in zebrafish are also correlated with conserved signatures in human tumor samples, and down-regulation of the B-myb signature genes is associated with retention of p53 function. Our findings show that zebrafish screens can uncover cancer pathways, and demonstrate that loss of function of bmyb is associated with cancer.cell cycle

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“…Adult male zebrafish of the AB strain were mutagenized with ethylnitrosourea and crossed to wild-type AB females in order to generate the F 1 heterozygous generation. F 2 haploid embryos were obtained by squeezing anesthetized F 1 females and fertilizing the eggs in vitro with UV-inactivated sperm (6). At 36 h postfertilization (hpf) haploids were collected and used for antiphosphorylated H3 (pH3) staining.…”

Section: Methodsmentioning

confidence: 99%

Topoisomerase IIα Is Required for Embryonic Development and Liver Regeneration in Zebrafish

Dovey

Patton

Bowman

et al. 2009

Molecular and Cellular Biology

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Topoisomerases solve the topological problems encountered by DNA throughout the lifetime of a cell. Topoisomerase II␣, which is highly conserved among eukaryotes, untangles replicated chromosomes during mitosis and is absolutely required for cell viability. A homozygous lethal mutant, can4, was identified in a screen to identify genes important for cell proliferation in zebrafish by utilizing an antibody against a mitosis-specific marker, phospho-histone H3. Mutant embryos have a decrease in the number of proliferating cells and display increases in DNA content and apoptosis, as well as mitotic spindle defects. Positional cloning revealed that the genetic defect underlying these phenotypes was the result of a mutation in the zebrafish topoisomerase II␣ (top2a) gene. top2a was found to be required for decatenation but not for condensation in embryonic mitoses. In addition to being required for development, top2a was found to be a haploinsufficient regulator of adult liver regrowth in zebrafish. Regeneration analysis of other adult tissues, including fins, revealed no heterozygous phenotype. Our results confirm a conserved role for TOP2A in vertebrates as well as a dose-sensitive requirement for top2a in adults.The accurate and complete replication and separation of chromosomes during mitosis is vital for the viability of cells. One potential complication encountered during cell division is the topological and tensional pressures put on DNA during these processes. Topoisomerases are responsible for enzymatically winding, unwinding, knotting, and unknotting reactions that are necessary for solving the topological problems of DNA. Genetic and biochemical studies in bacteria and yeast have revealed two classes of topoisomerases, type I and type II (38). Type I topoisomerases act on DNA by creating a temporary single-strand nick in DNA, passing the intact strand through the broken strand and then religating the nick. Type II topoisomerases, alternatively, function by binding to two double-stranded DNA molecules, generating a double-stranded break in one of the strands, passing the intact strand through the broken strand, and religating the broken strand.During DNA replication, sister chromatids become tangled with each other as a by-product of their duplication. The primary cellular function of type II topoisomerases during mitosis is the decatenation (untangling) of sister chromatids. Consistent with this essential role, genetic studies Saccharomyces cerevisiae have revealed that temperature-sensitive topoisomerase II mutants (TOP2) are unviable at nonpermissive temperatures (10,16,36). These mutants arrest during M phase due to their incompletely decatenated chromatids.Humans have two TOP2 homologues, topoisomerase II alpha (TOP2A) and topoisomerase II beta (TOP2B). Although they are both functionally redundant with the yeast homologue, TOP2A and TOP2B are genetically unique. As the products of different loci, TOP2A and TOP2B have different expression patterns. Specifically, TOP2A expression peaks during M phase, wh...

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Chapter 5 Production of Haploid and Diploid Androgenetic Zebrafish (Including Methodology for Delayed in Vitro Fertilization)

Cited by 17 publications

References 18 publications

Characterization andIn VitroControl of MPF Activity in Zebrafish Eggs

Characterization andIn VitroControl of MPF Activity in Zebrafish Eggs

A zebrafish bmyb mutation causes genome instability and increased cancer susceptibility

Topoisomerase IIα Is Required for Embryonic Development and Liver Regeneration in Zebrafish

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