Anti-diuretic activity of a CAPA neuropeptide can compromise Drosophila chill tolerance.

For insects, chilling injuries that occur in the absence of freezing are often related to a systemic loss of ion and water balance that leads to extracellular hyperkalemia, cell depolarization and the triggering of apoptotic signalling cascades. The ability of insect ionoregulatory organs (e.g. the Malpighian tubules) to maintain ion balance in the cold has been linked to improved chill tolerance, and many neuroendocrine factors are known to influence ion transport rates of these organs. Injection of micromolar doses of CAPA (an insect neuropeptide) have been previously demonstrated to improve Drosophila cold tolerance, but the mechanisms through which it impacts chill tolerance are unclear, and low doses of CAPA have been previously demonstrated to cause anti-diuresis in insects, including dipterans. Here, we provide evidence that low (femtomolar) and high (micromolar) doses of CAPA impair and improve chill tolerance, respectively, via two different effects on Malpighian tubule ion and water transport. While low doses of CAPA are anti-diuretic, reduce tubule K+ clearance rates and reduce chill tolerance, high doses facilitate K+ clearance from the haemolymph and increase chill tolerance. By quantifying CAPA peptide levels in the central nervous system, we estimated the maximum achievable hormonal titres of CAPA and found further evidence that CAPA may function as an anti-diuretic hormone in Drosophila melanogaster We provide the first evidence of a neuropeptide that can negatively affect cold tolerance in an insect and further evidence of CAPA functioning as an anti-diuretic peptide in this ubiquitous insect model.

Waking inactivity as a welfare indicator in laboratory mice: investigating postures, facial expressions and depression-like states.

Animal welfare assessment relies on valid and practical indicators of affect. In mice, the most widely used research vertebrates, lying still with eyes open, inactive-but-awake (IBA) in the home cage, has potential to be one such indicator. IBA is elevated in barren, conventional housing compared with well-resourced, enriched housing, and predicts immobility in Forced Swim Tests, a common measure of 'helplessness' in depression research. In Experiment 1, using females from three strains (C57BL/6, Balb/c and DBA/2), we first replicated past findings, confirming higher levels of IBA in conventional cages and a positive relationship between IBA and helplessness. We then extended this research to three other signs of depression: changes in weight and sleep, and reduced hippocampal volume. Here, IBA positively covaried with body mass index, with sleep in DBA/2s and conventionally housed BALB/cs, and negatively covaried with hippocampal volume in conventionally housed C57BL/6s. In Experiment 2, we sought to refine the phenotype of IBA to improve its accuracy as a welfare indicator. Here, scoring IBA performed in hunched postures appeared to improve its accuracy as an indicator in Balb/c mice. Additional research is now needed to further refine the phenotype of IBA and to confirm whether it reflects states consistent with depression, or instead other underlying poor welfare conditions.

Where are you from? Female mice raised in enriched or conventional cages differ socially, and can be discriminated by other mice.

Laboratory rodents raised in environmentally-enriched (EE) cages differ behaviourally and cognitively from conventionally-housed (CH) animals. We hypothesised that mice can detect such differences, testing this using differentially-raised female C57BL/6 s as subjects, and differentially-raised female BALB/cs and DBA/2 s as stimuli, in Social Approach Tests. Because more prone to signs of depression, anxiety, stereotypic behaviour (SB) and aggression, we further hypothesised that CH mice would be less sociable and socially attractive than EE mice. A novel familiarisation paradigm pre-exposed subjects to non-cagemate EE and CH stimulus mice before testing in Social Approach Tests. CH subjects proved less sociable than EE subjects: an effect unrelated to general exploration, anxiety or depression-like traits, and driven specifically by reduced interest in CH stimulus mice. Providing further evidence that CH and EE stimulus mice could be distinguished, subjects proved most attracted to mice from housing unlike their own. CH subjects thus preferred EE over CH stimulus mice, while EE subjects tended to prefer CH over EE: patterns that were not mediated by any measured aspect of stimulus mouse behaviour. Differential bodyweight also seemed unimportant, as was scent: soiled CH and EE bedding/nesting did not elicit the same discrimination. Instead, subjects who avoided CH stimulus mice and were attracted to EE stimulus mice were those who received the most agonism in their home cages. Together this provides the first demonstration that mice can distinguish between individuals raised in enriched or conventional cages, and suggests that receiving agonism from cagemates may motivate mice to seek new, less aggressive companions.