The Replica Set Method: A High-throughput Approach to Quantitatively Measure &lt;em&gt;Caenorhabditis elegans&lt;/em&gt; Lifespan

Cornwell, Adam; Llop, Jesse R.; Salzman, Peter; Thakar, Juilee; Samuelson, Andrew V.

doi:10.3791/57819-v

Cited by 3 publications

(10 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For replica set lifespan experiments ( Figure Supplement 3 ), animals were synchronized as previously described, followed by moving animals to 16°C during development (L1 – L4), after which FUdR was added, and animals were moved to 20 or 25 °C as indicated. Replica set lifespan was trials were setup, scored, and results were analyzed as described in (Cornwell et al, 2018; Cornwell and Samuelson, 2020).…”

Section: Methodsmentioning

confidence: 99%

“…Primary data for (D-F) can be found in Supplementary File 1 (this study). Lifespan experiments in (A-F) used the replicate set method (for detailed protocols see (Cornwell et al, 2018; Cornwell and Samuelson, 2020), which is as accurate and precise as traditional approaches to assess changes in C. elegans lifespan and have high resolution to detect differences (Cornwell et al, 2022): in daf-2(e1370);daf-16(mgDf47) the p-value comparisons (empty vector to hpk-1(RNAi) ) are significant at both 25 and 20°C, but result in small absolute decreases in median lifespan (0.68 and 1.12 days, respectively). Curves were plotted using WormLife (https://github.com/samuelsonlab-urmc/wormlife) (Cornwell et al, 2018).…”

Section: Figure Supplementmentioning

confidence: 99%

“…Lifespan experiments in (A-F) used the replicate set method (for detailed protocols see (Cornwell et al, 2018; Cornwell and Samuelson, 2020), which is as accurate and precise as traditional approaches to assess changes in C. elegans lifespan and have high resolution to detect differences (Cornwell et al, 2022): in daf-2(e1370);daf-16(mgDf47) the p-value comparisons (empty vector to hpk-1(RNAi) ) are significant at both 25 and 20°C, but result in small absolute decreases in median lifespan (0.68 and 1.12 days, respectively). Curves were plotted using WormLife (https://github.com/samuelsonlab-urmc/wormlife) (Cornwell et al, 2018). (G) Lifespan of C. elegans with enhanced sensitivity to RNAi within the nervous system ( unc-119p::sid-1; eri-1(mg366) ) treated with either empty vector (grey line) or daf-2(RNAi) (red line) or unc-119p::sid-1; eri-1(mg366); sur-5p::hpk-1 animals treated with daf-2(RNAi) (black line) using the Lifespan Machine method (Stroustrup et al, 2013).…”

Section: Figure Supplementmentioning

confidence: 99%

“…Lifespan graph is representative of one trial with two replicates. Curves were plotted using WormLife (https://github.com/samuelsonlab-urmc/wormlife) (Cornwell et al, 2018). See Supplementary File 1 for details and additional trial.…”

Section: Figure Supplementmentioning

confidence: 99%

See 3 more Smart Citations

Homeodomain-interacting protein kinase maintains neuronal homeostasis during normalCaenorhabditis elegansaging and systemically regulates longevity from serotonergic and GABAergic neurons

Lázaro-Peña

Cornwell

Díaz-Balzac

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Aging and the age-associated decline of the proteome is determined in part through neuronal control of evolutionarily conserved transcriptional effectors, which safeguard homeostasis under fluctuating metabolic and stress conditions by regulating an expansive proteostatic network. We have discovered the Caenorhabditis elegans homeodomain-interacting protein kinase (HPK-1) acts as a key transcriptional effector to preserve neuronal integrity, function, and proteostasis during aging. Loss of hpk-1 results in drastic dysregulation in expression of neuronal genes, including genes associated with neuronal aging. During normal aging hpk-1 expression increases throughout the nervous system more broadly than any other kinase. Within the aging nervous system, hpk-1 is co-expressed with key longevity transcription factors, including daf-16 (FOXO), hlh-30 (TFEB), skn-1 (Nrf2), and hif-1, which suggests hpk-1 expression mitigates natural age-associated physiological decline. Consistently, pan-neuronal overexpression of hpk-1 extends longevity, preserves proteostasis both within and outside of the nervous system, and improves stress resistance. Neuronal HPK-1 improves proteostasis through kinase activity. HPK-1 functions cell non-autonomously within serotonergic and GABAergic neurons to improve proteostasis in distal tissues by specifically regulating distinct components of the proteostatic network. Increased serotonergic HPK-1 enhances the heat shock response and survival to acute stress. In contrast, GABAergic HPK-1 induces basal autophagy and extends longevity. Our work establishes hpk-1 as a key neuronal transcriptional regulator critical for preservation of neuronal function during aging. Further, these data provide novel insight as to how the nervous system partitions acute and chronic adaptive response pathways to delay aging by maintaining organismal homeostasis.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Figure Supplementmentioning

confidence: 99%

Section: Figure Supplementmentioning

confidence: 99%

Section: Figure Supplementmentioning

confidence: 99%

See 2 more Smart Citations

Homeodomain-interacting protein kinase maintains neuronal homeostasis during normalCaenorhabditis elegansaging and systemically regulates longevity from serotonergic and GABAergic neurons

Lázaro-Peña

Cornwell

Díaz-Balzac

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Enough "replica" plates are set up at the outset of the experiment for each planned observation based on typical lifespan characteristics of the strains already established. Resulting data was fit to logistic function curve for determination of median lifespan, as previously described (Cornwell et al 2018(Cornwell et al , 2022.…”

Section: Lifespanmentioning

confidence: 99%

TheC. elegansMyc-family of transcription factors coordinate a dynamic adaptive response to dietary restriction

Cornwell,

Zhang,

Thondamal

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Dietary restriction (DR), the process of decreasing overall food consumption over an extended period of time, has been shown to increase longevity across evolutionarily diverse species and delay the onset of age-associated diseases in humans. InCaenorhabditis elegans, the Myc-family transcription factors (TFs) MXL-2 (Mlx) and MML-1 (MondoA/ChREBP), which function as obligate heterodimers, and PHA-4 (orthologous to forkhead box transcription factor A) are both necessary for the full physiological benefits of DR. However, the adaptive transcriptional response to DR and the role of MML-1::MXL-2 and PHA-4 remains elusive. We identified the transcriptional signature ofC. elegansDR, using theeat-2genetic model, and demonstrate broad changes in metabolic gene expression ineat-2DR animals, which requires bothmxl-2andpha-4. While the requirement for these factors in DR gene expression overlaps, we found many of the DR genes exhibit an opposing change in relative gene expression ineat-2;mxl-2animals compared to wild-type, which was not observed ineat-2animals withpha-4loss. We further show functional deficiencies of themxl-2loss in DR outside of lifespan, aseat-2;mxl-2animals exhibit substantially smaller brood sizes and lay a proportion of dead eggs, indicating that MML-1::MXL-2 has a role in maintaining the balance between resource allocation to the soma and to reproduction under conditions of chronic food scarcity. Whileeat-2animals do not show a significantly different metabolic rate compared to wild-type, we also find that loss ofmxl-2in DR does not affect the rate of oxygen consumption in young animals. The gene expression signature ofeat-2mutant animals is consistent with optimization of energy utilization and resource allocation, rather than induction of canonical gene expression changes associated with acute metabolic stress -such as induction of autophagy after TORC1 inhibition. Consistently,eat-2animals are not substantially resistant to stress, providing further support to the idea that chronic DR may benefit healthspan and lifespan through efficient use of limited resources rather than broad upregulation of stress responses, and also indicates that MML-1::MXL-2 and PHA-4 may have different roles in promotion of benefits in response to different pro-longevity stimuli.

show abstract

The C. elegans Myc-family of transcription factors coordinate a dynamic adaptive response to dietary restriction

Cornwell,

Zhang,

Thondamal

et al. 2024

GeroScience

View full text Add to dashboard Cite

The Replica Set Method: A High-throughput Approach to Quantitatively Measure <em>Caenorhabditis elegans</em> Lifespan

Cited by 3 publications

References 0 publications

Homeodomain-interacting protein kinase maintains neuronal homeostasis during normalCaenorhabditis elegansaging and systemically regulates longevity from serotonergic and GABAergic neurons

Homeodomain-interacting protein kinase maintains neuronal homeostasis during normalCaenorhabditis elegansaging and systemically regulates longevity from serotonergic and GABAergic neurons

TheC. elegansMyc-family of transcription factors coordinate a dynamic adaptive response to dietary restriction

The C. elegans Myc-family of transcription factors coordinate a dynamic adaptive response to dietary restriction

Contact Info

Product

Resources

About