biostudies-arrayexpressRyo Higuchi-SanabriaCaenorhabditis eleganshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15031Mild mitochondrial stress can produce positive effects, a phenomenon referred to as \"mitohormesis.\" This process involves activation of signaling pathways such as the mitochondrial unfolded protein response (UPRmt), which helps restore mitochondrial function and has also been linked to improved health and extended lifespan across various model organisms. In C. elegans, mitohormesis can be triggered through several means—including inhibition of the electron transport chain (ETC), reduction in mitochondrial protein translation, or impaired mitochondrial import—all of which can lead to UPRmt-mediated lifespan extension. However, not all triggers of UPRmt result in increased longevity. For instance, while inhibiting ETC complex II strongly activates UPRmt, it has not been associated with lifespan extension. These findings raise the possibility that UPRmt activation alone may not directly promote longevity. In this study, we aim to investigate this complexity by examining how different mitochondrial stressors that induce UPRmt influence the lifespan of C. elegans. We use RNA-sequencing to profile genome-wide transcriptional responses, with the goal of identifying transcriptomic patterns that may clarify the relationship between UPRmt and longevity.biostudies-arrayexpressLibrary Construction - Library construction was performed by Novogene using their standard pipeline.Growth Protocol - Standard bleaching protocol was used to synchronize animal populations. Animals were L1 arrested for a maximum of 16 hours to further synchronize populations. Animals were grown on RNAi indicated from L1 until Day 1 of adulthood and material was collected on Day 1.Nucleic Acid Extraction - After the final thaw, chloroform was added at a 1:5 chlorform/trizol ratio and aqueous separation of RNA was performed via centrifugation in a heavy gel phase-lock tube (VWR, 10847-802). The aqueous phase was mixed 1:1 with isopropanol then applied to a Qiagen RNeasy Mini Kit (74106) and RNA purification was performed as per manufacturer’s directions.Sample Collection - For RNA isolation, all RNa collection was performed at day 2 of adulthood. ~1000 animals were harvested from RNAi plates using M9. Animals were pelleted by gravity by allowing adult worms to settle to the bottom of the tube and aspirating off eggs and L1. Animals were washed and gravity settled 3x to remove a majority of progeny, then animals were placed into Trizol solution and worms were freeze/thawed 3x with liquid nitrogen with a 30 sec vortexing step between each freeze cycle.Sequencing - RNA-sequencing was performed by Novogene using their standard pipeline.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - Three biological replicates were measured per condition. Reads were trimmed with trim_galore-0.6.5-1 and mapped to WBcel235 with STAR-2.7.3a (Dobin et al. 2013). Mapped reads were counted to genes using feature Counts (Subread-2.0.0) (Liao, Smyth, and Shi 2019). Unwanted variations were removed with SVASeq-3.50.0 (Leek et al. 2014) using R-4.2.2.2 and differential expression analysis was performed with DESeq2-1.38.3 (Love, Huber, and Anders 2014).UnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000RNA-seq of coding RNACaenorhabditis elegansRyo Higuchi-SanabriafalseTranscriptomic analysis of mitohormesis associated with lifespan extension in Caenorhabditis elegansMild mitochondrial stress can produce positive effects, a phenomenon referred to as \"mitohormesis.\" This process involves activation of signaling pathways such as the mitochondrial unfolded protein response (UPRmt), which helps restore mitochondrial function and has also been linked to improved health and extended lifespan across various model organisms. In C. elegans, mitohormesis can be triggered through several means—including inhibition of the electron transport chain (ETC), reduction in mitochondrial protein translation, or impaired mitochondrial import—all of which can lead to UPRmt-mediated lifespan extension. However, not all triggers of UPRmt result in increased longevity. For instance, while inhibiting ETC complex II strongly activates UPRmt, it has not been associated with lifespan extension. These findings raise the possibility that UPRmt activation alone may not directly promote longevity. In this study, we aim to investigate this complexity by examining how different mitochondrial stressors that induce UPRmt influence the lifespan of C. elegans. We use RNA-sequencing to profile genome-wide transcriptional responses, with the goal of identifying transcriptomic patterns that may clarify the relationship between UPRmt and longevity.2025-08-31T00:00:00Z2025-08-31T01:02:29.506Z2025-04-15T12:17:39.865ZE-MTAB-15031ERP171649EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003816EFO_0003738EFO_0004184