Nuclear Transplants from Adult Somatic Cells Generated by a Novel Method Using Diploidized Eggs as Recipients in Medaka Fish (<i>Oryzias latipes</i>)

Bubenshchikova, Ekaterina; Kaftanovskaya, Elena M.; Hattori, Manabu; Kinoshita, Masato; Adachi, Tomoko; Hashimoto, Hisashi; Ozato, Kenjiro; Wakamatsu, Yuko

doi:10.1089/clo.2008.0014

Cited by 13 publications

(8 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Ploidy estimation was performed using methods described elsewhere 16. Briefly, either part of the caudal fin from an adult fish or a whole recently hatched larva was minced in extraction buffer (CyStain DNA 2step, Partec GmbH, Germany) and incubated for 10 to 20 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

“…In the transfer of embryonic cell nuclei to enucleated eggs, transplants have been generated in cyprinid fish 7-10, loach, Misgurnus fossilis 11, medaka, Oryzias latipes 12 and zebrafish, Danio rerio 13, 14. However, until our recent work using newly established methods in the small freshwater fish model, medaka 15, 16, nuclear transfer using adult somatic cell nuclei had not yet been reported. In those studies, we used non-enucleated and diploidized eggs as recipients for the transfer of adult somatic cell nuclei and consistently obtained diploid and fertile transplants that originated from the donor nuclei.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Nuclear Transfer of Embryonic Cell Nuclei to Non-enucleated Eggs in Zebrafish, Danio rerio

Hattori¹,

Hashimoto²,

Bubenshchikova³

et al. 2011

Int. J. Biol. Sci.

Self Cite

View full text Add to dashboard Cite

We previously established a novel method for nuclear transfer in medaka (Oryzias latipes) using non-enucleated, diploidized eggs as recipients for adult somatic cell nuclei. Here we report the first attempt to apply this method to another fish species. To examine suitability of using non-enucleated eggs as recipients for nuclear transfer in the zebrafish (Danio rerio), we transferred blastula cell nuclei from a wild-type donor strain to non-enucleated, unfertilized eggs from a golden recipient strain. As a result, 31 of 184 (16.8%) operated eggs developed normally and reached the adult stage. Twenty-eight (15.2%) of these transplants showed wild-type phenotype and the remaining three (1.6%) were golden. Except for one individual that exhibited diploid/tetraploid mosaicism, all of the wild-type nuclear transplants were either triploid or diploid. While all of 19 triploid transplants were infertile, a total of six transplants (21.4%) were fertile (five of the eight diploid transplants and one transplant exhibiting ploidy mosaicism). Except for one diploid individual, all of the fertile transplants transferred both the wild-type golden gene allele (slc24a5) as well as the phenotype, the wild-type body color, to their F1 and F2 progeny in a typical Mendelian fashion. PCR analysis of slc24a5 suggested that triploidy originated from a fused nucleus in the diploid donor and haploid recipient nuclei, and that the sole origin of diploidy was the diploid donor nucleus. The results of the present study demonstrated the suitability of using non-enucleated eggs as recipients for nuclear transfer experiments in zebrafish.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Nuclear Transfer of Embryonic Cell Nuclei to Non-enucleated Eggs in Zebrafish, Danio rerio

Hattori¹,

Hashimoto²,

Bubenshchikova³

et al. 2011

Int. J. Biol. Sci.

Self Cite

View full text Add to dashboard Cite

show abstract

“…For RFX3 mRNA detection, the PCR primers 5′-CGTTGCGCAGATATACGTCAC-3′ (sense) and 5′-AGGCGTCTCTCCAGTAGCTTG-3′ (antisense) were used. EF-1a mRNA detection was conducted as previously described [68]. (B) Northern blot analysis pc/glis3 mRNA was detected in WT kidney but not in pc kidney or WT liver by northern blot analysis using the fragment of glis3 cDNA containing exons 3–6 as a probe.…”

Section: Supporting Informationmentioning

confidence: 99%

Polycystic Kidney Disease in the Medaka (Oryzias latipes) pc Mutant Caused by a Mutation in the Gli-Similar3 (glis3) Gene

et al. 2009

Self Cite

View full text Add to dashboard Cite

Polycystic kidney disease (PKD) is a common hereditary disease in humans. Recent studies have shown an increasing number of ciliary genes that are involved in the pathogenesis of PKD. In this study, the Gli-similar3 (glis3) gene was identified as the causal gene of the medaka pc mutant, a model of PKD. In the pc mutant, a transposon was found to be inserted into the fourth intron of the pc/glis3 gene, causing aberrant splicing of the pc/glis3 mRNA and thus a putatively truncated protein with a defective zinc finger domain. pc/glis3 mRNA is expressed in the epithelial cells of the renal tubules and ducts of the pronephros and mesonephros, and also in the pancreas. Antisense oligonucleotide-mediated knockdown of pc/glis3 resulted in cyst formation in the pronephric tubules of medaka fry. Although three other glis family members, glis1a, glis1b and glis2, were found in the medaka genome, none were expressed in the embryonic or larval kidney. In the pc mutant, the urine flow rate in the pronephros was significantly reduced, which was considered to be a direct cause of renal cyst formation. The cilia on the surface of the renal tubular epithelium were significantly shorter in the pc mutant than in wild-type, suggesting that shortened cilia resulted in a decrease in driving force and, in turn, a reduction in urine flow rate. Most importantly, EGFP-tagged pc/glis3 protein localized in primary cilia as well as in the nucleus when expressed in mouse renal epithelial cells, indicating a strong connection between pc/glis3 and ciliary function. Unlike human patients with GLIS3 mutations, the medaka pc mutant shows none of the symptoms of a pancreatic phenotype, such as impaired insulin expression and/or diabetes, suggesting that the pc mutant may be suitable for use as a kidney-specific model for human GLIS3 patients.

show abstract

“…The mechanism leading to the elimination of the oocyte nuclei is still unknown (Bubenshchikova et al, 2008).…”

Section: Scnt Of Other Speciesmentioning

confidence: 99%

Further Advice on the Implications of Animal Cloning (SCNT)

2009

EFS2

View full text Add to dashboard Cite

Nuclear Transplants from Adult Somatic Cells Generated by a Novel Method Using Diploidized Eggs as Recipients in Medaka Fish (Oryzias latipes)

Cited by 13 publications

References 33 publications

Nuclear Transfer of Embryonic Cell Nuclei to Non-enucleated Eggs in Zebrafish, Danio rerio

Nuclear Transfer of Embryonic Cell Nuclei to Non-enucleated Eggs in Zebrafish, Danio rerio

Polycystic Kidney Disease in the Medaka (Oryzias latipes) pc Mutant Caused by a Mutation in the Gli-Similar3 (glis3) Gene

Further Advice on the Implications of Animal Cloning (SCNT)

Contact Info

Product

Resources

About