Acute Smurf mortality and inter-phase dependence in Drosophila and mice identified through comprehensive modelling and statistical analysis of two-phase ageing

Abstract

Ageing is traditionally conceived as a continuous process of progressive physiological decline. However, recent evidence across species suggests that ageing may instead proceed through distinct phases. Using state-of-the-art statistical methods, we develop a rigorous analysis of longitudinal survival data from 1,159 individually tracked female Drosophila melanogaster. This data-driven analysis leads us to introduce a new parametric model of transition rates within the two-phase ageing framework. Flies were monitored using the Smurf assay, which detects increased intestinal permeability through leakage of an ingested blue dye, and is a strong biological marker of ageing. The Smurf phenotype identifies a sharp transition from a non-Smurf state to a Smurf state that precedes death. Our results yield three key findings. First, the Smurf transition rate follows a Gompertz-Makeham law, increasing exponentially with age. Second, contrary to previous constant-rate assumptions, newly transitioned Smurf flies exhibit remarkably high mortality - approximately 40% die within 24 hours - followed by an exponential decline in death rate that stabilises to a lower constant baseline. Third, we identified a mild but statistically significant negative dependence between time spent non-Smurf and subsequent Smurf lifespan. Our best-fit model captures a potential bimodal nature of mortality curves using simple, biologically interpretable functions. Validation using data from two mouse strains confirms the broader applicability of this framework. These results establish a quantitative foundation for the two-phase ageing paradigm and highlight a critical period of vulnerability immediately following the physiological transition to frailty.