Lack of Genetic Variation in Low Heterozygosity and Clonal Animals Creates Lack of Physiological Variation

Data variability frequently complicates reproducibility and interpretation of experimental results. Such variability arises from numerous sources such as differences in procedures or not accounting for key biological factors (e.g. sex, biological rhythms, prandial state). Making the situation more problematic, variation in physiological performance is often viewed as highly labile, easily and rapidly influenced by environmental stressors, development, etc., making it diffcult to pin down a source for variation. Undeniably, however, unknown (or ignored) genetic variation among and within strains/lines can also be a significant source of data variability in published physiological measurements, though surprisingly this has not been extensively investigated as a specific source of physiological variation. We hypothesized that variation in physiological performance is correlated with the intrinsic degree of genetic variability of the subject animal. To test this hypothesis, we employed two animal models: 1) Inbred lines ( e.g., NHGRI-1) derived from wild type strains of the zebrafish ( Danio rerio), with an estimated 15% of the genetic heterozygosity of wild type AB zebrafish, and 2) the parthenogenetically reproducing marbled crayfish ( Procambarus virginalis), all specimens of which are genetically identical clones. For these two animal models, we measured both physiological variables (e.g. heart rate, stroke volume, cardiac output, oxygen consumption) and morphological variables (e.g. yolk-chorion ratio, body mass, embryo mass, total length, condition factor, specific growth rate) during development. We subjected the two animal models to environmental stress in the form of both temperature and hypoxia to stimulate physiological responses that could be compared and contrasted among populations. From these data we then calculated the resultant coeffcients of variation for measured variables for wild type and low/zero heterozygosity populations and/or species. In zebrafish, both the wildtype AB and NHRGI-1 lines showed similar developmental trajectories characterized by similar mean values for physiological and morphological variables. Additionally, similar mean values for physiological and morphological variables were recorded in the face of temperature and hypoxia challenge. Yet, importantly, the coeffcient of variation for each measured variable was significantly lower in NHGRI-1 than AB larvae for >90% of the assessed endpoints. In the clonal crayfish, genetically identical early stage marbled crayfish reared in different temperatures or oxygen levels show major acclimation responses, but generally showed less morphological and physiological variation about the mean than sexually-reproducing species crayfish with inherently much greater genetic variation, as evident from comparisons of calculated coeffcients of variation. A key question regarding the clonal crayfish is how can there be any morphological or physiological variation between individuals? We suggest that variability that persists may arise from microenvironmental differences during rearing (e.g. egg position during incubation on the mother’s pleon) and/or stochastic differences in gene expression (e.g. due to random epimutations) in this clonal species. In conclusion, genetic diversity clearly contributes to physiological variability. For future experiments, low heterozygosity lines and/or clonal species may be useful for decreasing inter-individual variation, thus aiding interpretation of results and enhancing reproducibility. In any event, scientific documentation of physiological studies should include as much information on (genetic) background of the experimental animals as possible. NSF IOS-2103499. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.