C. elegans Collagen Deposition Assessment Service

Extracellular matrices (ECM) exist within cells and tissues, playing crucial roles in tissue integrity, geometry and function. In mammals, the major component of the ECM is collagen, which makes up about 30% of the total protein mass in the body. The ECM integrity declines with a continuous reduction of collagen mass induced by the accumulation of damage from collagen fragmentation, glycation, and oxidation during aging. Many age-dependent diseases, including cardiovascular diseases, chronic liver diseases, diabetes, and cancer are implicated in the deterioration of ECM integrity. Meanwhile, chronic inflammation, accumulation of molecular damage and injury during aging may lead to abnormal collagen deposition resulting in fibrosis. In general, as the function of age, the overall collagen mass tends to decline. Here we provide the assessment service of collagen deposition in C. elegans to help customers to explore novel insights in maintenance and remodelling of the ECM during aging.

A proper model system for studying collagen deposition

As a powerful model organism, the nematode C. elegans is evidently a good option to insight to collagen deposition and remodelling during aging. Previous studies revealed that collagen synthesis in C. elegans declines during aging. Specifically, most long-lived C. elegans mutants delay this progressive reduction in collagen mass. And it seems essential to healthy aging and longevity in C. elegans through actively prolonging the biosynthesis of collagens during their lifetime. In addition, we have listed the additional unique advantages of a well-established aging model C. elegans to explore this phenomenon during aging.

  • The well-established genetics;
  • The short lifespan (about 3 weeks) of the nematodes;
  • The transparent body allows to directly monitor ECM homeostasis and integrity through tagged ECM components with fluorescent proteins in vivo.

Service offerings in Creative Biogene

Assessment of collagen deposition in C. elegans

As a histology method, the Herovici staining is used to assess the extent of collagen deposition in tissues. In contrast with other commonly used collagen staining protocols, the Herovici technique can distinguish young and mature collagen without cross polarized filters, exhibiting an obvious advantage in the visualization and analysis of processes. Note that young collagen is stained blue, mature collagen appears pink to red and cytoplasm is counterstained yellow when using a sequential polychromatic staining. Here we provide the service to stain the collagen and distinguish between young and mature collagen ratios in C. elegans using the Herovici staining combining with freeze-cracking method.

Herovici-stained C. elegans at day 1 of adulthood.Herovici-stained C. elegans at day 1 of adulthood. (Teuscher, A. C., et al, 2019)

C. elegans collagen quantification

Collagens have characteristic [Gly-X-Y] repeats. The prolonged stretches of which play crucial roles in forming a stable triple helix from the three constitutive collagen chains. Given that collagens are enriched with hydroxyproline, a posttranslational modification is not common in other proteins. Here we provide the service to quantify the total collagen level of C. elegans through measuring hydroxyproline abundancy. Total collagen quantification is combined with total collagen determination and a colorimetric method to estimate collagen levels for C. elegans is adapted.

Collagen quantification using hydroxyproline assay for C. elegans.Collagen quantification using hydroxyproline assay for C. elegans. (Teuscher, A. C., et al, 2019)

Applications of C. elegans collagen deposition assessment service

  • Assessing collagen deposition during aging and in age-related pathologies
  • Exploring the maintenance and remodelling of the ECM during aging

As a dedicated provider of C. elegans model services, Creative Biogene has developed a systematic platform, containing a histological and colorimetric method to reveal the alteration of collagen deposition in C. elegans from visual and quantified perspective. With the strict experimental procedure, and experienced technicians, we are confident to serve our customers with high quality. Additionally, our services are tailored to meet customers’ specific needs. To improve research outcomes and the speed of success, please don’t hesitate to contact us for more information.

Reference

  1. Teuscher, A. C., et al. (2019). "Assessing Collagen Deposition During Aging in Mammalian Tissue and in Caenorhabditis elegans." Methods in molecular biology (Clifton, N.J.), 1944, 169-188.

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