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Garcia Martinez, A.

Publications and source records attributed to Garcia Martinez, A..

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TailTimer: an open-source device for automating the rodent tail immersion assay

The tail immersion assay is a widely used method for measuring acute thermal pain in a way which is quantifiable and reproducible. It is non-invasive and measures response to a stimulus that may be encountered by an animal in its natural environment. However, tail withdrawal latency data are usually collected manually, and precise temperatures of the water at the time of measurement are most often not recorded. These two factors can reduce the reproducibility of tail immersion assay data. We designed a device, TailTimer, which uses the Raspberry Pi single-board computer and a temperature sensor, to automatically record both tail withdrawal latency and water temperature. The device has a radio frequency identification (RFID) system that can record the ID of animals. Our software recognizes several specific RFID keys as user interface commands, which allows TailTimer to be operated via RFID fobs. We also programmed the device to only allow tests to be conducted when the water is within {+/-} 0.25 {degrees}C of the target temperature. Data recorded using the TailTimer device showed a linear relationship between tail withdrawal latency and water temperature when tested between 47 - 50 {degrees}C. We also observed a profound effect of water mixing speed on tail withdrawal latency. Our data further revealed significant strain and sex differences, valorizing TailTimer in its ability to detect genetically-determined variations in thermal pain sensitivity. Significance StatementQuantification of tail withdrawal latency in response to thermal pain has essentially remained the same since the method was first introduced decades ago and relies on manual recording of water temperature and tail withdrawal latency. Such manual methods engender relatively substantial variability and are potential contributors to some of the discrepancies present among relevant research. The open source TailTimer device we report here is simple and inexpensive to manufacture. The RFID-based user interface is ergonomic, especially in animal facilities where space is limited and gloves are mandatory. We anticipate that the increased reproducibility of tail withdrawal latency provided by TailTimer will augment its utility in nociception and addiction research.

animal behavior and cognition

Reversing age: dual species measurement of epigenetic age with a single clock

Young blood plasma is known to confer beneficial effects on various organs in mice. However, it was not known whether young plasma rejuvenates cells and tissues at the epigenetic level; whether it alters the epigenetic clock, which is a highly-accurate molecular biomarker of aging. To address this question, we developed and validated six different epigenetic clocks for rat tissues that are based on DNA methylation values derived from n=593 tissue samples. As indicated by their respective names, the rat pan-tissue clock can be applied to DNA methylation profiles from all rat tissues, while the rat brain-, liver-, and blood clocks apply to the corresponding tissue types. We also developed two epigenetic clocks that apply to both human and rat tissues by adding n=850 human tissue samples to the training data. We employed these six clocks to investigate the rejuvenation effects of a plasma fraction treatment in different rat tissues. The treatment more than halved the epigenetic ages of blood, heart, and liver tissue. A less pronounced, but statistically significant, rejuvenation effect could be observed in the hypothalamus. The treatment was accompanied by progressive improvement in the function of these organs as ascertained through numerous biochemical/physiological biomarkers and behavioral responses to assess cognitive functions. Cellular senescence, which is not associated with epigenetic aging, was also considerably reduced in vital organs. Overall, this study demonstrates that a plasma-derived treatment markedly reverses aging according to epigenetic clocks and benchmark biomarkers of aging.

developmental biology