Development of inter-family nuclear transplant embryos by transplanting the nuclei from the loach blastulae into the non-enucleated zebrafish eggs

2003 ◽  
Vol 21 (1) ◽  
pp. 64-71 ◽  
Author(s):  
Li Li ◽  
Zhang Shicui ◽  
Yuan Jinduo ◽  
Li Hongyan
Keyword(s):  
Nuclear Transplant

Relationship between kinetics of nuclear swelling and subsequent in vitro development of nuclear transplant rabbit embryos

1992 ◽  
Vol 37 (1) ◽  
pp. 185 ◽  
Author(s):  
P.G. Adenot ◽  
Y. Heyman ◽  
P. Chesné ◽  
V.H. Rao ◽  
J.P. Renard
Keyword(s):  
In Vitro Development ◽  
Nuclear Transplant ◽  
Vitro Development ◽  
Kinetics Of ◽  
Rabbit Embryos

scholarly journals Nuclear transplant studies on the reduction in numbers of presumptive germ nuclei in exconjugants of Paramecium caudatum

1982 ◽  
Vol 56 (1) ◽  
pp. 453-460
Author(s):  
K. Mikami
Keyword(s):  
Vegetative Cell ◽  
Posterior Region ◽  
Anterior Region ◽  
Nuclear Transplantation ◽  
Paramecium Caudatum ◽  
Vegetative Phase ◽  
Nuclear Transplant ◽  
Nuclear Differentiation

Nuclear differentiation in exconjugants of Paramecium caudatum is closely associated with a brief localization of the postzygotic nuclei near the opposite ends of the cell, with the germinal nucleus (micronucleus) in the anterior region and the somatic nuclei (macronuclei) in the posterior region. The posterior nuclei cannot regenerate to produce micronuclei when all four anterior nuclei are removed. There is no difference among the anterior four presumptive micronuclei, because, when any three of them were removed, the remaining nucleus was able to divide at each postconjugational fission and to persist as a micronucleus during the vegetative phase. This conclusion agrees with the results of transplanting a presumptive micronucleus into a vegetative cell. Cells during the vegetative phase, however, normally have only one micronucleus. Micronuclear number must be reduced to arrive at the uni-micronucleate condition after the stage of macro- and micronuclear differentiation. Elimination of supernumerary presumptive micronuclei, which had been indicated by morphological observations, was confirmed by the results of nuclear transplantation studies.

scholarly journals Nuclear transfer from teratocarcinoma cells into mouse oocytes and eggs

Development ◽  
1990 ◽  
Vol 108 (2) ◽  
pp. 337-348
Author(s):  
J.A. Modlinski ◽  
D. Gerhauser ◽  
B. Lioi ◽  
H. Winking ◽  
K. Illmensee
Keyword(s):  
Embryonal Carcinoma ◽  
Preimplantation Embryos ◽  
Time Intervals ◽  
Electrophoretic Variants ◽  
Glucose Phosphate ◽  
Teratocarcinoma Cells ◽  
Nuclear Transplant ◽  
Ec Cells

A spontaneous ovarian teratocarcinoma was isolated from a LT/Sv mouse female and converted into an ascites tumor from which embryonal carcinoma (EC) cells were dissociated. Non-enucleated and enucleated, activated oocytes were fused with EC cells and either cultured in vitro or transferred into ligated oviducts of Swiss/A females. The nucleocytoplasmic hybrids cultured in vitro up to 22 h were examined cytologically at various time intervals. EC nuclei showed morphological remodelling in the foreign cytoplasm. EC chromosomes and female pronuclear chromosomes together formed a common metaphase. The nucleocytoplasmic hybrids developed in vivo were analyzed cytologically between the first and third day after oviduct transfer. The majority of embryos developed abnormally and, in a few instances, they had passed several cleavage divisions and reached, at best, a developmental stage resembling a premature morula. Fertilized, enucleated eggs were fused with EC cells or microinjected with EC nuclei. The resulting nucleocytoplasmic hybrids were either cultured in vitro or in vivo up to the fourth day. Enzyme tests were carried out on the nuclear transplant embryos, using electrophoretic variants of glucose phosphate isomerase (GPI) in order to distinguish between EC nuclei (GPI-A) and recipient eggs (GPI-B). The EC-specific GPI could be detected in about one third of the embryos analyzed and, in several instances, also together with the egg-specific GPI. Most of them were arrested during early cleavage divisions. Some embryos cleaved abnormally or mimicked normal embryogenesis. In a few instances, development resulted in embryos that resembled late preimplantation embryos.

96 TRICHOSTATIN A IMPROVED THE QUALITY OF RABBIT NUCLEAR TRANSFER EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 165 ◽  
Author(s):  
J. Xu ◽  
L.-Y. Sung ◽  
J. Zhang ◽  
X. Tian ◽  
Y. E. Chen ◽  
...  
Keyword(s):  
Histone Acetylation ◽  
Trichostatin A ◽  
Donor Cell ◽  
Cumulus Cells ◽  
Cell Number ◽  
Developmental Potential ◽  
Number Of Factors ◽  
Nuclear Transplant ◽  
Blastocyst Quality ◽  
And Control

Nuclear reprogramming is dependent upon a number of factors, including chromatin organization and modification. Trychostatin A (TSA), a histone deacetylase inhibitor, was used to increase histone acetylation and to improve reprogrammability in both cattle and mice. The objective of the study was to determine whether TSA could improve the pre-implantational development potential of rabbit nuclear transplant (NT) embryos. Rabbit oocytes were flushed from the oviducts of superovulated donors treated with the regime of FSH and hCG. Cumulus cells were then denuded from the oocytes by incubation in 0.5% hyaluronidase and pipetting. Oocyte enucleation was conducted in 10% FBS M199 and confirmed under fluorescence microscopy. Cumulus cells were prepared as nuclear donors for NT; a donor cell with the diameter approximately 15–19 µm was transferred into the perivitelline space of an enucleated oocyte, and subsequently fused with the oocyte recipient by application of 3 direct current pulses at 3.2 kV cm−1 for a duration of 20 µs/pulse. Fused embryos were activated by the same electrical stimulation regime described above, and subsequently cultured in M199 + 10% FBS containing 2.0 mM 6-dimethylaminopurine (DMAP) and 5 µg mL−1 cycloheximide for 1 h. Rabbit NT embryos were cultured in 5 nM TSA-2.5% FBS-B2 medium for 10 h before being transferred into regular medium (FBS-B2). The TSA-treated embryos (5 nM vs. 0 nM) were cultured in 400 µL FBS-B2 medium for 5 days in 5% CO2 in a humidified atmosphere at 38.5°C (initiation of activation = Day 0). NT embryo development to cleaved (2 to 4 cell), morula, and blastocyst stages was evaluated on Day 2, Day 3, and Day 5, respectively. The selected NT blastocysts were counted for cell numbers following Hoechst 33342 epifluorescenin staining. The results (Table 1) showed that there was no difference on pre-implantational development of cloned embryos between TSA-added and control groups (P > 0.05). However, a significantly higher cell number per NT blastocyst was found in the TSA-added group (357 vs. 113; P < 0.05). This indicated that the blastocyst quality in NT embryos was improved with the addition of TSA by increasing histone acetylation activity. The developmental potential of TSA-treated NT embryos to term is under investigation. Table 1.Effects of TSA on the pre-implantational development of cloned rabbit embryos This work was supported by NIH/NCRR-SBIR grant: 1R43RR020261-01.

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

Zygote ◽  
2009 ◽  
Vol 18 (1) ◽  
pp. 33-39 ◽  
Author(s):  
M. Pérez-Camps ◽  
J. Cardona-Costa ◽  
M. Francisco-Simao ◽  
F. García-Ximénez
Keyword(s):  
Danio Rerio ◽  
Egg Activation ◽  
Technical Approach ◽  
Animal Pole ◽  
Technical Aspects ◽  
Nuclear Transplant ◽  
Donor Nucleus ◽  
Adult Cell ◽  
Definition Of ◽  
Adult Cells

SummaryZebrafish somatic nuclear transplant has only been attempted using preactivated eggs. In this work, three methods to carry out the nuclear transplant using adult cells before, during and after the egg activation/fertilization were developed in zebrafish with the aim to be used in reprogramming studies. The donor nucleus from somatic adult cells was inserted: (method A) in the central region of the egg and subsequently fertilized; (method B) in the incipient animal pole at the same time that the egg was fertilized; and (method C) in the completely defined animal pole after fertilization. Larval and adult specimens were obtained using the three methods. Technical aspects related to temperature conditions, media required, egg activation/fertilization, post-ovulatory time of the transplant, egg aging, place of the donor nucleus injection in each methodology are presented. In conclusion, the technical approach developed in this work can be used in reprogramming studies.

scholarly journals Relationship between Nuclear Remodeling and Development in Nuclear Transplant Rabbit Embryos1

1991 ◽  
Vol 45 (3) ◽  
pp. 455-465 ◽  
Author(s):  
Philippe Collas ◽  
James M. Robl
Keyword(s):  
Nuclear Transplant ◽  
Nuclear Remodeling

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

Zygote ◽  
2011 ◽  
Vol 20 (3) ◽  
pp. 277-280
Author(s):  
J. Cardona-Costa ◽  
M. Pérez-Camps ◽  
F. García-Ximénez
Keyword(s):  
Somatic Cell ◽  
Central Region ◽  
Somatic Cells ◽  
Cell Reprogramming ◽  
Oocyte Activation ◽  
Animal Pole ◽  
Preliminary Results ◽  
Somatic Cell Reprogramming ◽  
Nuclear Transplant

SummarySomatic 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.

Producing multiple generations of bovine nuclear transplant embryos

1991 ◽  
Vol 35 (1) ◽  
pp. 273 ◽  
Author(s):  
S.L. Stice ◽  
C.L. Keefer ◽  
M. Maki-Laurila ◽  
P.E. Phillips
Keyword(s):  
Multiple Generations ◽  
Nuclear Transplant
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