Nathalie Chenais

• French National Institute for Agriculture, Food, and Environment (INRAE)

Xenopus egg extract is a powerful material to modify cultured cells fate and to induce cellular reprogramming in mammals. In this study, the response of goldfish fin cells to in vitro exposure to Xenopus egg extract, and subsequent culture, was studied using a cDNA microarray approach, gene ontology and KEGG pathways analyses, and qPCR validation....

Somatic cell reprogramming in vitro prior to nuclear transfer is one strategy expected to improve clone survival during development. In this study, we investigated the reprogramming extent of fish fin somatic cells after in vitro exposure to Xenopus egg extract and subsequent culture. Using a cDNA microarray approach, we observed drastic changes in...

Somatic cell reprogramming in vitro prior to nuclear transfer is one strategy expected to improve clone survival during development. In this study, we investigated the reprogramming extent of fish fin somatic cells after in vitro exposure to Xenopus egg extract and subsequent culture. Using a cDNA microarray approach, we observed drastic changes in...

Nuclear transfer consists in injecting a somatic nucleus carrying valuable genetic information into a recipient oocyte to sire a diploid offspring which bears the genome of interest. It requires that the oocyte (maternal) DNA is removed. In fish, because enucleation is difficult to achieve, non-enucleated oocytes are often used and disappearance of...

Nuclear transfer consists in injecting a somatic nucleus carrying valuable genetic information into a recipient oocyte to sire a diploid offspring who bears the genome of interest. It requires that the oocyte (maternal) DNA is removed. In fish, because enucleation is difficult to achieve, non-enucleated oocytes are often used and disappearance of t...

Reprogramming of cultured cells using Xenopus egg extract involves controlling four major steps: plasma membrane permeabilization, egg factors import into the nucleus, membrane resealing, and cell proliferation. Using propidium iodide to assess plasma membrane permeability, we established that 90% of the cultured fin cells were permeabilized by dig...

Cryopreservation of genetic resources is of major interest for the security of biodiversity and the sustainability of the agronomic industry. Cryopreservation of somatic cells is one means of preserving the genome of both parents, but regeneration of functional breeders requires the mastering of somatic cell NT (SCNT). In mammals, SCNT consists of...

Prions induce a fatal neurodegenerative disease in infected host brain based on the refolding and aggregation of the host-encoded prion protein PrPC into PrPSc. Structurally distinct PrPSc conformers can give rise to multiple prion strains. Constrained interactions between PrPC and different PrPSc strains can in turn lead to certain PrPSc (sub)popu...

Supplementary Figures, Supplementary Tables, Supplementary Notes and Supplementary References

Somatic cells bear a major potential as cell resource for biodiversity conservation: they carry the genome of both parents, and they can be collected whatever the age or sex of the donor. In fish, cryobanking of somatic cells is highly valuable because eggs and embryos cannot be cryopreserved. Then, the sole use of cryopreserved sperm for animal re...

The development of fin primary cell cultures for in vitro cellular and physiological studies is hampered by slow cell outgrowth, low proliferation rate, poor viability, and sparse cell characterization. Here, we investigated whether the recycling of fresh explants after a first conventional culture could improve physiological stability and sustaina...

Domestication is a long-term process during which wild fish are acclimated to farming conditions and hopefully are reproduced over several generations, possibly using selective breeding. Preservation of the genetic diversity of the original population, together with that of the ongoing selection steps, is important for ecological and economical pur...

In naturally acquired transmissible spongiform encephalopathies, the pathogenic agents or prions spread from the sites of initial peripheral uptake or replication to the brain where they cause progressive and fatal neurodegeneration. Routing via the peripheral nervous system is considered as one of the main pathways to attain the central nervous sy...

L’utilisation du kit CyQuant® NF pour des comptages cellulaires à haut débit et pour des mesures de proliférations a été testée sur des cellules de nageoires de poisson rouge issues de culture primaire, comme une alternative aux lourds comptages manuels sur cellules de Malassez. Nous montrons que le dosage présente, pour nos cellules, une très bonn...

Nuclear transfer has the potential to become one strategy for fish genetic resources management, by allowing fish reconstruction from cryopreserved somatic cells. Survival rates after nuclear transfer are still low however. The part played by unsuitable handling conditions is often questioned, but the different steps in the procedure are difficult...

The development of biotechnologies such as nuclear transfer in goldfish (Carassius auratus) requires that the spawns be available all year round. For this reason, an off-season maturation procedure was set up in our laboratory. The aim of this study was to describe how these artificially maturated females responded to induced ovulation, and how egg...

The development of nuclear transfer in Goldfish (Carassius auratus) requires egg handling to be optimized 1) to maintain the ability of the egg to withstand embryo development 2) to allow micromanipulation in order to enucleate the egg and inject the donor nucleus 3) to favour egg cytoplasm ability for nucleus reprogramming. In this work, several e...

Spatial and temporal control of ovine prion protein (Prnp) gene expression was achieved in mice using two transgenes: a Prnp minigene with tet-operator sequences inserted 5' to exon 1 and a mouse neurofilament genomic clone carrying the chimeric-repressor TRSID cDNA. In bi-transgenic mice, ovine PrP(C) expression could be reversibly controlled in n...

Synopsis Prions are unconventional agents of proteic nature that are formed of abnormal conformations of the host-encoded prion protein (PrP). They cause fatal neurodegenerative diseases in both animals and humans, and can be transmitted between species as exemplified in humans by the emergence of variant Creutzfeldt-Jakob disease following the epi...

Description of the Bovine PrP Transgenic Mice (tg540 Line) (92 KB DOC)