Loss of intestinal nuclei and intestinal integrity in aging C. elegans
Matthew D. McGee
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorDarren Weber
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorNicholas Day
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorCathy Vitelli
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorDanielle Crippen
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorLaura A. Herndon
Albert Einstein College of Medicine, Center for C. elegans Anatomy, 1410 Pelham Parkway South, Rm 601, Bronx, NY 10461, USA
Search for more papers by this authorDavid H. Hall
Albert Einstein College of Medicine, Center for C. elegans Anatomy, 1410 Pelham Parkway South, Rm 601, Bronx, NY 10461, USA
Search for more papers by this authorSimon Melov
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorMatthew D. McGee
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorDarren Weber
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorNicholas Day
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorCathy Vitelli
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorDanielle Crippen
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorLaura A. Herndon
Albert Einstein College of Medicine, Center for C. elegans Anatomy, 1410 Pelham Parkway South, Rm 601, Bronx, NY 10461, USA
Search for more papers by this authorDavid H. Hall
Albert Einstein College of Medicine, Center for C. elegans Anatomy, 1410 Pelham Parkway South, Rm 601, Bronx, NY 10461, USA
Search for more papers by this authorSimon Melov
Buck Institute for Age Research, 8001 Redwood Blvd, Novato, CA 94945, USA
Search for more papers by this authorSummary
The roundworm C. elegans is widely used as an aging model, with hundreds of genes identified that modulate aging (Kaeberlein et al., 2002. Mech. Ageing Dev.123, 1115–1119). The development and bodyplan of the 959 cells comprising the adult have been well described and established for more than 25 years (Sulston & Horvitz, 1977. Dev. Biol.56, 110–156; Sulston et al., 1983. Dev. Biol.100, 64–119.). However, morphological changes with age in this optically transparent animal are less well understood, with only a handful of studies investigating the pathobiology of aging. Age-related changes in muscle (Herndon et al., 2002. Nature419, 808–814), neurons (Herndon et al., 2002), intestine and yolk granules (Garigan et al., 2002. Genetics161, 1101–1112; Herndon et al., 2002), nuclear architecture (Haithcock et al., 2005. Proc. Natl Acad. Sci. USA102, 16690–16695), tail nuclei (Golden et al., 2007. Aging Cell6, 179–188), and the germline (Golden et al., 2007) have been observed via a variety of traditional relatively low-throughput methods. We report here a number of novel approaches to study the pathobiology of aging C. elegans. We combined histological staining of serial-sectioned tissues, transmission electron microscopy, and confocal microscopy with 3D volumetric reconstructions and characterized age-related morphological changes in multiple wild-type individuals at different ages. This enabled us to identify several novel pathologies with age in the C. elegans intestine, including the loss of critical nuclei, the degradation of intestinal microvilli, changes in the size, shape, and cytoplasmic contents of the intestine, and altered morphologies caused by ingested bacteria. The three-dimensional models we have created of tissues and cellular components from multiple individuals of different ages represent a unique resource to demonstrate global heterogeneity of a multicellular organism.
Supporting Information
Fig. S1 Heterogeneity in old worms.
Fig. S2 Wild-type microvilli in young adults.
Fig. S3 Long-lived daf-2 worms have protected nuclear morphology.
Movie S1 4-day-old wild-type worm. All aligned methylene blue/pararosaniline cross sections from a 4-day-old wild-type worm.
Movie S2 20-day-old wild-type worm. All aligned methylene blue/pararosaniline cross sections from a 20-day-old wild-type worm. Anatomical features of interest are labeled.
Movie S3 The Intestinal lumen changes with age. Segmentation of the intestinal lumen (blue) from aligned methylene blue/pararosaniline cross sections (Movies S1 and S2) from a 4-day-old and 20-day-old wild-type worm. Cuticle is yellow. Small gaps in the intestinal lumen are due to imperfect image alignment from shifted or distorted sections.
Movie S4 Intestinal nuclei are lost with age. Nuclei from a 4-day-old and a 20-day-old wild-type worm. Surface models were created from DAPI staining. Blue nuclei are not annotated, red nuclei are proximal germline masses, yellow nuclei are sperm, and light blue nuclei are intestinal nuclei. Partway through the video, all but the intestinal nuclei ‘fall away’. The 20-day-old intestinal nuclei shift to magenta partway through to allow distinction between the 4-day-old intestinal nuclei.
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