Definition of three somatic adult cell nuclear transplant methods in zebrafish (<i>Danio rerio</i>): before, during and after egg activation by sperm fertilization

Pérez-Camps, M.; Cardona-Costa, J.; Francisco-Simão, M.; García-Ximénez, F.

doi:10.1017/s0967199409990049

Cited by 3 publications

(3 citation statements)

References 23 publications

(33 reference statements)

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“…Until now, in zebrafish, these events (parthenogenetic development) have only been possible via genetically inactive sperm (Nüsslein-Volhard and Dahm, 2002). However, the activation stimulus proposed here could serve for improving the embryo developmental rates in procedures which involve nuclear injection techniques in zebrafish such as NT (Huang et al, 2003;Siripattarapravat et al, 2009b;Pérez-Camps et al, 2009) or ICSI (Poleo et al, 2001 as occurs in mammals (Macháty, 2006). Moreover, in fish, the electrical (and/or chemical) stimuli would be the only ones available in these procedures once the egg has already been activated by water.…”

Section: Discussionmentioning

confidence: 99%

“…To our knowledge, the use of egg electroactivation in fish nuclear transplant (NT) has only been carried out on medaka fish (Bubenshchikova et al, 2007;Wakamatsu, 2008). In zebrafish, the NT of somatic nucleus is able to induce embryo development when activation is only promoted by water, usually reaching the MBT stage (Huang et al, 2003;Pérez-Camps et al, 2009;Siripattarapravat et al, 2009b).…”

Section: Resumen Electroactivación De Oocitos De Pez Cebra (Danio Rerio)mentioning

confidence: 99%

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Electroactivation of zebrafish (Danio rerio) eggs

Cardona-Costa

Pérez-Camps

García-Ximénez

1970

Span. j. agric. res.

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The zebrafish, has recently acquired a complex role in many emerging areas of knowledge, mainly in biomedical research (Nüsslein-Volhard and Dahm, 2002), becoming a new scientific tool in multitude of labs around the world. In this sense, several and varied are the topical areas where the zebrafish has established as a both efficient and capable vertebrate model for developing knowledge. As Streisinger (1986) pointed out, the reasons were their many useful biological characteristics.In zebrafish, egg activation takes place after they are discharged from the ovarian stroma and come into AbstractIn zebrafish, initial egg activation by water after being discharged from the ovarian stroma is followed by normal cleavages only in inseminated eggs. When sperm (genetically active or not) is not used as activating agent, reproductive techniques (as either nuclear transplant or intracytoplasmic sperm injection are ineff icient. In this work, three experiments of egg activation by electric stimuli were performed: In the first, six activation treatments were compared (Voltage × Pulses: 2.76 × 1; 2.76 × 2; 2.76 × 3 and 5.40 × 1; 5.40 × 2; 5.40 × 3). The group 5.40 × 3 showed the best results (32% activated). In the second experiment, an electrical treatment of 20 min was carried out. It consisted in a sequence of three equal electrical stimuli every 10 min (of 1 or 3 consecutive direct current square pulses for 20 µs each and applied at two voltage levels, 2.76 V or 5.4 V). It was observed that the number of pulses negatively affected the rates of damaged and lysed eggs. Moreover, only the 20 min treatment with the combination of 3 consecutive pulses at 2.76 V showed significant differences with their respective control group (43% vs 18% activated eggs, p < 0.05). In the third experiment, negative effects of egg ageing were observed. In conclusion, the best activation treatment for intact (non manipulated) zebrafish eggs concerns sequence B3. The electro-activation stimulus proposed here would be the only one available once the egg has already been activated by water.Additional key words: egg activation; fish; nuclear transplant; oocyte. Resumen Electroactivación de oocitos de pez cebra (Danio rerio)En pez cebra, una vez que los oocitos son descargados del estroma ovárico, su activación inicial por contacto en agua da lugar a divisiones normales únicamente cuando son fertilizados. Cuando no se utiliza semen (inactivado genéticamente o no), las técnicas reproductivas (como transplante nuclear o inyección espermática intracitoplasmática) son ineficientes. En el presente trabajo se desarrollaron tres experimentos de activación oocitaria por estímulos eléctricos: en el primero de ellos, se compararon seis tratamientos de activación (Voltaje × Pulsos: 2,76 × × 1; 2,76 × 2; 2,76 × 3 y 5,40 × 1; 5,40 × 2; 5,40 × 3). El grupo 5,4 × 3 mostró los mejores resultados (32% de activación). En el segundo experimento, se evaluó un tratamiento eléctrico de 20 min de duración. El tratamiento consistió en la aplicación de una secuencia de tre...

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Section: Discussionmentioning

confidence: 99%

Section: Resumen Electroactivación De Oocitos De Pez Cebra (Danio Rerio)mentioning

confidence: 99%

Electroactivation of zebrafish (Danio rerio) eggs

Cardona-Costa

Pérez-Camps

García-Ximénez

1970

Span. j. agric. res.

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“…The two somatic cell nuclear transplant techniques, as well as the equipment employed were described by Pérez-Camps et al . (2010a).…”

Section: Methodsmentioning

confidence: 99%

Somatic cells derived from haploid larvae are feasible as donors for nuclear transplant in zebrafish. Preliminary results

Cardona-Costa

Pérez-Camps

García-Ximénez

2011

Zygote

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Summary Somatic cells derived from zebrafish haploid larval (both androgenetic and gynogenetic) cultures were used as donors for nuclear transplant into non-enucleated oocytes. Nuclei were transplanted either before or simultaneously with oocyte activation in the central region and in the incipient animal pole, respectively. Against expected results, 20% of transplanted embryos during oocyte activation using cells of gynogenetic origin reached the 100% epiboly stage, even two survived for up to 5 days, whereas no development was observed when cells from androgenetic origin were used. Results derived from this work open a novel possibility of studying somatic cell reprogramming and imprinting phenomena in zebrafish.

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