ABSTRACT
BACKGROUND: Over the last decade a number of species, from farm animals to rodents, have been cloned using somatic cell nuclear transfer technology (SCNT). This technique has the potential to revolutionize the way that genetically modified animals are made. In its current state, the process of SCNT is very inefficient (<5% success rate), with several technical and biological hurdles hindering development. Yet, SCNT provides investigators with powerful advantages over other approaches, such as allowing for prescreening for the desired level of transgene expression and eliminating the excess production of undesirable wild-type animals. The rat plays a significant role in biomedical research, but SCNT has been problematic for this species. In this study, we address one aspect of the problem by evaluating methods of activation in artificially constructed rat embryos. PRINCIPAL FINDINGS: We demonstrate that treatment with a calcium ionophore (ionomycin) combined with a variety of cyclin-dependent kinase inhibitors is an effective way to activate rat embryos. This is in contrast to methods developed for the mouse embryo, which tolerates much less specific chemical treatments. Methods developed to activate mouse embryos do not translate well to rat embryos. CONCLUSIONS: Activation methods developed for one species will not necessarily translate to another species, even if it is closely related. Further, the parthenogenic response to chemical activators is not always a reliable indicator of how reconstructed embryos will react to the same activation method. A better understanding of rat oocyte physiology, although essential for developing better models of disease, may also provide insights that will be useful for making the SCNT process more efficient.
Subject(s)
Cloning, Organism/methods , Cyclin-Dependent Kinases/antagonists & inhibitors , Developmental Biology/methods , Gene Expression Regulation, Developmental , Nuclear Transfer Techniques , Protein Kinase Inhibitors/pharmacology , Animals , Calcium/metabolism , Female , Gene Expression Regulation, Enzymologic , Ionomycin/pharmacology , Ionophores , Oocytes/cytology , Rats , Rats, Long-EvansABSTRACT
Plants will play an essential role in providing life support for any long-term space exploration or habitation. We are evaluating the feasibility of an adaptable system for measuring the response of plants to any unique space condition and optimizing plant performance under those conditions. The proposed system is based on a unique combination of systems including the rapid advances in the field of plant genomics, microarray technology for measuring gene expression, bioinformatics, gene pathways and networks, physiological measurements in controlled environments, and advances in automation and robotics. The resulting flexible module for monitoring and optimizing plant responses will be able to be inserted as a cassette into a variety of platforms and missions for either experimental or life support purposes. The results from future plant functional genomics projects have great potential to be applied to those plant species most likely to be used in space environments. Eventually, it will be possible to use the plant genetic assessment and control system to optimize the performance of any plant in any space environment. In addition to allowing the effective control of environmental parameters for enhanced plant productivity and other life support functions, the proposed module will also allow the selection or engineering of plants to thrive in specific space environments. The proposed project will advance human exploration of space in the near- and mid-term future on the International Space Station and free-flying satellites and in the far-term for longer duration missions and eventual space habitation.
Subject(s)
Ecological Systems, Closed , Genes, Plant , Life Support Systems , Plant Development , Space Flight , Weightlessness , Agriculture/methods , Agriculture/trends , Automation , Biotechnology , Environment, Controlled , Gene Expression Regulation, Plant , Humans , Plant Physiological Phenomena , Plants/genetics , Plants/metabolism , RoboticsABSTRACT
A novel presenilin 1 mutation, insR352, associated with a frontal temporal dementia phenotype has been identified (E. A. Rogaeva et al., 2001, Neurology 57, 621-625). This mutation does not increase Abeta42 levels, but instead acts as dominant negative presenilin, decreasing amyloid beta protein (Abeta) production by inhibiting gamma-secretase cleavage of the Abeta precursor. The distinct clinical phenotype associated with this mutation suggests that chronic partial inhibition of gamma-secretase activity may result in neurodegeneration.