Autofluorescence as a measure of senescence in C. elegans: look to red, not blue or green

Pincus, Zachary; Mazer, Travis C; Slack, Frank J.

doi:10.18632/aging.100936

Cited by 82 publications

(98 citation statements)

References 26 publications

(57 reference statements)

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“…Second, we assessed age-associated tissue deterioration through quantitative measurements of textural order and disorder in brightfield images (Johnston et al, 2008; Pincus et al, 2011). Third, we measured age-related declines in homeostatic ability, as manifested in the accumulation of fluorescent non-hydrolyzable materials in intestinal endosomes (Klass, 1977; Clokey and Jacobson, 1986; Gerstbrein et al, 2005; Hermann et al, 2005; Pincus et al, 2016). Fourth, we evaluated nutritional history and somatic investment through cross-sectional body size (Pincus et al, 2011; Hulme et al, 2010).…”

Section: Resultsmentioning

confidence: 99%

Extended Twilight among Isogenic C. elegans Causes a Disproportionate Scaling between Lifespan and Health

et al. 2016

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Summary Although many genetic factors and lifestyle interventions are known to affect the mean lifespan of animal populations, the physiological variation displayed by individuals across their lifespans remains largely uncharacterized. Here, we use a custom culture apparatus to continuously monitor five aspects of aging physiology across hundreds of isolated Caenorhabditis elegans individuals kept in a constant environment from hatching until death. Aggregating these measurements into an overall estimate of senescence, we find two chief differences between longer- and shorter-lived individuals. First, though long- and short-lived individuals are physiologically equivalent in early adulthood, longer-lived individuals experience a lower rate of physiological decline throughout life. Second, and counter-intuitively, long-lived individuals have a disproportionately extended “twilight” period of low physiological function. While longer-lived individuals experience more overall days of good health, their proportion of good to bad health, and thus their average quality of life, is systematically lower than that of shorter-lived individuals. We conclude that within a homogeneous population reared under constant conditions, the period of early-life good health is comparatively uniform and the most plastic period in the aging process is end-of-life senescence.

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Section: Resultsmentioning

confidence: 99%

Extended Twilight among Isogenic C. elegans Causes a Disproportionate Scaling between Lifespan and Health

et al. 2016

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“…During aging, the C. elegans autofluorescence increases (Pincus et al , 2016), making it more difficult to score levels and localization of GFP-fused proteins. Figure 8 shows transgenic animals that express a GFP-tagged HIM-4 fusion, which is localized to the basement membrane, either imaged with standard filter sets or with the triple band filter sets (on day 1 and day 8 of adulthood).…”

Section: Discussionmentioning

confidence: 99%

“…The source of the autofluorescence, whether it is lipofuscin, AGE, tryptophan metabolites, or something else, is still unclear. However, this autofluorescence increases during aging and the two main tissues that show the highest autofluorescence are the intestine and the uterus in C. elegans (Pincus et al , 2016). With current fluorescent filter sets (TRITC, DAPI, FITC), three different autofluorescent wavelengths have been characterized in C. elegans .…”

Section: Introductionmentioning

confidence: 99%

“…With current fluorescent filter sets (TRITC, DAPI, FITC), three different autofluorescent wavelengths have been characterized in C. elegans . The red autofluorescence (visualized by TRITC) progressively increases with age, the blue autofluorescence (visualized by DAPI) peaks right before death, and the green autofluorescence (visualized by FITC) is a mixture from the red and blue autofluorescence (Pincus et al , 2016). …”

Section: Introductionmentioning

confidence: 99%

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Overcoming Autofluorescence to Assess GFP Expression During Normal Physiology and Aging in Caenorhabditis elegans

Teuscher

Ewald

2018

BIO-PROTOCOL

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Green fluorescent protein (GFP) is widely used as a molecular tool to assess protein expression and localization. In C. elegans, the signal from weakly expressed GFP fusion proteins is masked by autofluorescence emitted from the intestinal lysosome-related gut granules. For instance, the GFP fluorescence from SKN-1 transcription factor fused to GFP is barely visible with common GFP (FITC) filter setups. Furthermore, this intestinal autofluorescence increases upon heat stress, oxidative stress (sodium azide), and during aging, thereby masking GFP expression even from proximal tissues. Here, we describe a triple band GFP filter setup that separates the GFP signal from autofluorescence, displaying GFP in green and autofluorescence in yellow. In addition, yellow fluorescent protein (YFP) remains distinguishable from both the yellowish autofluorescence and GFP with this triple band filter setup. Although some GFP intensity might be lost with the triple band GFP filter setup, the advantage is that no modification of currently used transgenic GFP lines is needed and these GFP filters are easy to install. Hence, by using this triple band GFP filter setup, the investigators can easily distinguish autofluorescence from GFP and YFP in their favorite transgenic C. elegans lines.

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“…56 Exposure time remained constant throughout the experiment. These images show that the GFP signal peaks in early adulthood (around three days post-hatch), while intestinal autofluorescence builds up slowly over time.…”

Section: Figurementioning

confidence: 99%

A simple culture system for long-term imaging of individual C. elegans

et al. 2017

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We have miniaturized standard culture techniques to rear arrays of isolated, individual C. elegans throughout their lives on solid gel media. The resulting apparatus is compatible with brightfield and fluorescent microscopy, enabling longitudinal studies of morphology and fluorescent transgene expression. Our culture system exploits a novel crosslinking reaction between a polyethylene glycol hydrogel and a silicone elastomer to constrain animals to individual “corrals” on the gel surface. These devices are simple to construct on the benchtop with commercially available reagents, and, unlike microfluidic isolation methods, does not rely on micropatterned materials. We demonstrate that this new culture method has negligible effects on the physiology of C. elegans compared to standard culture on agar plates. In addition, RNAi techniques are effective in this system. Finally, the hydrogel–silicone binding chemistry that we developed also allows traditional microfluidic devices to be covalently attached to gel substrates instead of glass.

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Autofluorescence as a measure of senescence in C. elegans: look to red, not blue or green

Cited by 82 publications

References 26 publications

Extended Twilight among Isogenic C. elegans Causes a Disproportionate Scaling between Lifespan and Health

Extended Twilight among Isogenic C. elegans Causes a Disproportionate Scaling between Lifespan and Health

Overcoming Autofluorescence to Assess GFP Expression During Normal Physiology and Aging in Caenorhabditis elegans

A simple culture system for long-term imaging of individual C. elegans

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