Abstract
Several steps of sturgeon somatic cell nuclear transfer (SCNT) have been recently established, but improvements are needed to make it a feasible tool to preserve the natural populations of this group of endangered species. The donor cell position inside the recipient egg seems to be crucial for its reprogramming; therefore by injecting multiple donor somatic cells instead of a single cell with a single manipulation, we increased the potential for embryo development. Using the Russian sturgeon Acipenser gueldenstaedtii as a multiple cell donor and sterlet Acipenser ruthenus as the non-enucleated egg recipient, we obtained higher proportion of eggs developing into embryos than previously reported with single-SCNT. Molecular data showed the production of a specimen (0.8%) contained only the donor genome with no contribution from the recipient, while two specimens (1.6%) showed both recipient and donor genome. These findings are the first report of donor DNA integration into a sturgeon embryo after interspecific cloning. In all, we provide evidence that cloning with the multiple donor somatic cells can be feasible in the future. Despite the fact that the sturgeon cloning faces limitations, to date it is the most promising technique for their preservation.
Highlights
The Acipenseridae is an ancient family that faces internal and external threats including the loss of species genetic integrity through frequent interspecific hybridization[1], habitat degradation, and overfishing for their roe processed into caviar[2]
To evaluate the effect of deep donor cell injection in the central region of the recipient egg, we monitored development resulting from intraspecific and interspecific single-Somatic cell nuclear transfer (SCNT)
Initial cleavage was shown in the transplants and shallow injection in the animal pole of the recipient egg was performed in the subsequent experiments
Summary
The Acipenseridae is an ancient family that faces internal and external threats including the loss of species genetic integrity through frequent interspecific hybridization[1], habitat degradation, and overfishing for their roe processed into caviar[2]. Utilizing a single fin cell harvested from an albino sterlet, Acipenser ruthenus and Russian sturgeon, Acipenser gueldenstaedtii transplanted into non-enucleated eggs from sterlet achieved low development rate of reconstructed NTs, of 18.1% and 12%, respectively[11]. The newly developed cloning technique increases the potential for the donor fin cell to be placed in the most favorable position inside the recipient sterlet egg and reprogrammed. Females and males Russian sturgeon are reproductively mature at 10–16 and 8–13 years, respectively[20], and beluga at 15–18 and 10–15 years, respectively[21] These species are categorized as critically endangered[3], and fin tissue is an excellent source of donor genomic material, as the harvesting does not cause irreparable damage to the fish[10,22]. The use of sterlet as a recipient species and beluga as donor species has benefits for molecular genotyping using recently developed species-specific primers allowing routine identification of the NTs origin[24]
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