Extreme Life: Microvolume analytics to probe animal adaptation to environmental stress

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My lab focuses on biochemical adaptation to discover the molecular mechanisms that support animal survival of extreme environmental stresses (e.g. cold & freezing temperatures, oxygen limitation, dehydration). Our main models include frogs that survive whole body freezing, small mammals that hibernate for months, and turtles that can live underwater for weeks without breathing oxygen. We explore both unique and unifying molecular characteristics that aid survival, particularly the mechanisms of metabolic rate depression that permit prolonged life extension under stress conditions by lowering animal energy needs to just a fraction of normal. This work not only extends our understanding of how metabolism is regulated, and the limits to which it can be stretched, but has multiple potential biomedical applications; e.g. for tissue/organ cryopreservation or the development of inducible torpor as a treatment option. Our current research program has four main directions: (a) the regulatory roles of microRNAs in directing the fate of gene transcripts and suppressing protein translation during hypometabolism, (b) the action of epigenetic mechanisms (e.g. histone modifications, DNA methylation) in stress-responsive control of gene transcription, (c) the multi-faceted actions of posttranslational modifications to enzymes in adapting intermediary metabolism to achieve stress tolerance and prolonged hypometabolism, and (d) the validation and interpretation of “big data” from our recent involvements in genome, transcriptome, and proteome projects. Much of our research now derives from tiny tissue samples (<100 mg) and depends on qPCR quantification of mRNA and microRNA, highly accurate pipetting for preparation of PCR, enzymatic and Luminex assays in tiny volumes, and precise quantification of nucleotide and protein concentrations. The present application requests a 3-instrument package to increase capacity and automation of our workflow: (1) a qPCR thermocycler that uses 384-well microplates, (2) a liquid handling robot for accurate pipetting of tiny volumes into microplates, and (3) a micro-volume spectrophotometer for DNA/RNA/protein quantification in 1-2 µL amounts. This package will greatly improve the research output of all HQP (including 9 PhD & 3 MSc students), support novel collaborative opportunities such as our recent work on primate torpor in lemurs, and allow us to maximize data acquisition from small samples via the use of micro-analytic methods.