Analysis of locomotor activity during LD.

Circadian rhythms are prevalent on Earth and temporally organize behaviour and physiology of organisms to occur in species-specific ‘temporal niches’. However, species differ in how strictly individuals are controlled by their circadian clock, suggesting that it may offer a selective advantage for an individual to extend its temporal niche under certain circumstances, for example during stressful environmental conditions. A potential mechanism controlling temporal niche adherence involves the evolutionary capacitor and chaperon protein HSP90, known to assist the proper folding of important signalling molecules. If HSP90 becomes rate limiting (e.g., under environmental stress) hidden genetic variation will be expressed, producing novel and potentially beneficial phenotypes for the individual. While this role of HSP90 is well established for morphological traits, we show here that it extends to regulation of temporal behavioural patterns. We show that within a small subset of clock neurons in the fly brain, HSP83, the fly homologue of HSP90, mitigates inter-individual behavioural variability. We provide evidence for the requirement of HSP83 for efficient transcription of the gene encoding the circadian neuropeptide Pigment Dispersing Factor (PDF), and for correct PDF accumulation in central clock neurons. Strikingly, Hsp83 mutants affect synchronized oscillations of the clock protein PERIOD (PER) in subsets of circadian clock neurons in the same way as flies without PDF, further supporting a role of Hsp83 in regulating Pdf. Our findings therefore provide a mechanistic explanation for HSP83 function in regulation of behavioural variability, and offer an explanation for how to restrict temporal niche extension to stressful environmental conditions.