The recent rise in speed and efficiency of new sequencing technologies have
facilitated high-throughput sequencing, assembly and analyses of genomes, advancing
ongoing efforts to analyze genetic sequences across major vertebrate groups.
Standardized procedures in acquiring high quality DNA and RNA and establishing cell
lines from target species will facilitate these initiatives. We provide a legal and
methodological guide according to four standards of acquiring and storing tissue for
the Genome 10K Project and similar initiatives as follows: four-star (banked
tissue/cell cultures, RNA from multiple types of tissue for transcriptomes, and
sufficient flash-frozen tissue for 1 mg of DNA, all from a single individual);
three-star (RNA as above and frozen tissue for 1 mg of DNA);
two-star (frozen tissue for at least 700 μg of DNA); and
one-star (ethanol-preserved tissue for 700 μg of DNA or less of
mixed quality). At a minimum, all tissues collected for the Genome 10K and other
genomic projects should consider each species’ natural history and follow
institutional and legal requirements. Associated documentation should detail as much
information as possible about provenance to ensure representative sampling and
subsequent sequencing. Hopefully, the procedures outlined here will not only
encourage success in the Genome 10K Project but also inspire the adaptation of
standards by other genomic projects, including those involving other biota.
In nondomestic and endangered species, the use of domestic animal oocytes as recipients for exotic donor nuclei causes the normal pattern of cytoplasmic inheritance to be disrupted, resulting in the production of nuclear-cytoplasmic hybrids. Evidence suggests that conflict between nuclear and cytoplasmic control elements leads to a disruption of normal cellular processes, including metabolic function and cell division. This study investigated the effects of nuclear-cytoplasmic interactions on the developmental potential of interspecies embryos produced by in vitro fertilization and somatic cell nuclear transfer: cattle x cattle, gaur x cattle, hybrid x cattle. Cattle control and hybrid embryos were examined for development to the blastocyst stage and blastocyst quality, as determined by cell number and allocation, apoptosis incidence, and expression patterns of mitochondria-related genes. These analyses demonstrated that a 100% gaur nucleus within a domestic cattle cytoplasmic environment was not properly capable of directing embryo development in the later preimplantation stages. Poor blastocyst development accompanied by developmental delay, decreased cell numbers, and aberrant apoptotic and related gene expression profiles, all signs of disrupted cellular processes associated with mitochondrial function, were observed. Developmental potential was improved when at least a portion of the nuclear genome corresponded to the inherited cytoplasm, indicating that recognition of cytoplasmic components by the nucleus is crucial for proper cellular function and embryo development. A better understanding of the influence of the cytoplasmic environment on embryonic processes is necessary before interspecies somatic cell nuclear transfer can be considered a viable alternative for endangered species conservation.
Cortisol is a hormone released when animals experience stress. We validated the measurement of cortisol from hair for use in wildlife using wild chipmunks, and tested the use of hair cortisol by measuring cortisol from chipmunks captured in natural and logged sites.
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