Research Paper Volume 12, Issue 12 pp 12268—12284
Infertility induced by auxin in PX627 Caenorhabditis elegans does not affect mitochondrial functions and aging parameters
- 1 Institute of Nutritional Sciences, Laboratory for Nutrition in Prevention and Therapy, Biomedical Research Center Seltersberg (BFS), Justus Liebig University Giessen, Giessen 35392, Germany
- 2 Molecular Nutrition Research, Interdisciplinary Research Center, Justus Liebig University Giessen, Giessen 35392, Germany
Received: April 1, 2020 Accepted: May 1, 2020 Published: June 8, 2020
https://doi.org/10.18632/aging.103413How to Cite
Copyright © 2020 Dilberger et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Caenorhabditis elegans is widely used for aging studies. 5-Fluoro-2´-deoxyuridine (FUdR) is commonly used to control offspring. While larvae are stopped from further development, also mitochondrial DNA and function may be affected. Since mitochondria and longevity are closely related, the use of FUdR may falsify possible studies. PX627, an auxin inducible infertility strain to control offspring, allows mitochondrial investigations during senescence without FUdR toxicity.
Longevity and health parameters were assessed in 2- and 10-day old nematodes wild-type N2 and PX627 treated with FUdR or auxin, respectively. Mitochondrial membrane potential, energetic metabolites and reactive oxygen species levels, were determined. mRNA expression levels of key genes involved were quantified using quantitative real-time PCR.
FUdR significantly increased lifespan and health parameters, as well as, mitochondrial function compared to untreated controls and auxin treated PX627. Although a decrease in all parameters could be observed in aged nematodes, this was less severe after FUdR exposure. Glycolysis was significantly up-regulated in aged PX627 compared to N2. Expression levels of daf-16, sir-2.1, aak-2, skn-1, atp-2 and atfs-1 were regulated accordingly.
Hence, auxin in PX627 might be a good alternative to control progeny, for mitochondrial- and longevity-related investigations in nematodes.
Abbreviations
ΔΨm: mitochondrial membrane potential; AID: auxin inducible degradation; ANOVA: one-way analysis of variance; aak-2: AMP-Activated Kinase; act-2: actin; ama-1: amanitin resistant; ATP: adenosine triphosphate; atfs-1: Activating Transcription Factor associated with Stress; atp-2: ATP synthase subunit; C. elegans: Caenorhabditis elegans; cDNA: copy deoxyribonucleic acid; DNA: deoxyribonucleic acid; daf-16: abnormal DAuer Formation; E. coli: Escherichia coli; FCCP: Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone; FUdR: 5-fluoro-2´-deoxyuridine; mtDNA: mitochondrial deoxyribonucleic acid; NGM: nematode growth medium; NIH: National Institute of Health; PCR: polymerase chain reaction; qRT-PCR: quantitative real-time polymerase chain reaction; Rh123: rhodamine 123; RNA: ribonucleic acid; ROS: reactive oxygen species; SD: standard deviation; sir-2.1: yeast SIR related; skn-1: SKiNhead; SWB: swelling buffer; UPRmt: mitochondrial unfolded protein response.