Alkalinization of a Single Pair of Glial Cells Increases Lifespan, Health Span, and Stress Resistance in C. elegans

Aging depends on genetic and environmental factors, but the specific cell types and mechanisms that coordinate aging of the entire organism are not yet fully understood. Recent studies in C. elegans revealed that glial cells serve as upstream regulators of aging by conveying information about their stress status to other tissues. Glial cells regulate ionic homeostasis, which is essential for neuronal function and survival. Here we investigated the role of glial ion channel CLH-1, which is a glial pH regulator, in aging. We found that loss of clh-1 extends lifespan, improves stress resistance, reduces neuronal damage, and extends health span. These effects are linked to protective pathways, including those for oxidative stress and autophagy, and depend on pH regulation in glia. A single pool of approximately 2000 synchronized worms from wildtype(N2), clh-1, daf-16, daf-16;clh-1, and clh-1;Amsh clh-1 rescue strains was grown to Day 1 of adulthood. All samples were collected in three biological replicates. Pelleted worms were lysed using a pellet pestle motor (Kontes) in TRIzol reagent (Thermo Fisher, Waltham, MA). Total RNA was isolated with the Nucleospin RNA kit (Macherey-Nagel) following the manufacturer’s protocol. The quality of the mRNA was assessed using a NanoDrop 1000 v3.8 (Thermo Scientific), and samples containing 300 ng/µL of RNA with a volume of 20 µL were sent for next-generation RNA sequencing.