Aging is a complicated degenerative process that is receiving increasing attention in recent years. Apart from age-related morphological change and behavioral change, the biochemical change is key aging biomarkers indicative of the physiological status of aging. The age-related biochemical change contains the generation of fluorescent compounds, the level of DNA damage, the content of protein carbonylation, the amount of yolk protein in the body cavity, metabolic activity, and enzymatic activities. Here we offer services to determine the above biochemical changes in an excellent model system C. elegans to help our customers to evaluate the treatment with factors, such as mutations, drugs, diet, and environmental conditions that influence aging.
Lipofuscin, a heterogeneous mixture of oxidized and cross-linked macromolecules, and advanced glycosylation end products are generated by sugar addition to free amino groups of proteins and subsequent cross-linking. They belong to fluorescent compounds, and the spectral properties of which are characterized by an excitation maximum of about 340nm and an emission maximum of about 430nm. Previous studies have revealed the changes of these fluorescent compounds with age and the relationship with other factors. The fluorescent material can be accelerated by the high levels of insulin/IGF signaling and dietary intake, and it is positively correlated with the decline in body movement. Here we provide a quantitative measurement service to measure fluorescent compounds, which can be visualized as granules and mainly located in but not restricted to the intestine.
In the worm's germline, DNA displays an age-related increase in the number of single-strand breaks and the levels of 5-methylcytosine, and along with the decline in the ability of DNA to function as a template for transcription in vitro with age. Protein carbonylation is one of the specific measurements of the macromolecules damage, which is caused by oxygen free radicals. There is a negative correlation between the level of protein carbonylation and the lifespan, suggesting that high levels of protein carbonylation may contribute to a shortened lifespan. Yolk protein is primarily synthesized in the intestine and later accumulates in oocytes and embryos in young adult hermaphrodites. While the yolk protein largely accumulates in the body cavity in older hermaphrodites, and is visualized as lipid-like droplets under immunoelectron microscopy, accumulating evidence coming from studies suggests that the accumulation of yolk protein in the body cavity is caused by the continued synthesis and secretion of yolk protein in the intestine after the time when oocyte production declines.
Measurements of metabolic activity, including carbon dioxide generation, oxygen consumption, heat production, ATP levels, and the amount of superoxide anion have been demonstrated to display age-related changes. Carbon dioxide generation by populations of live worms declines about 50% from day 6 to day 12, while oxygen consumption of populations of wild-type worms declines about 60% from adult day 0 to day 12. Heat produced by populations of live worms is a measure of the rate of catabolic processes, and declines about 80% from adult day 0 to day 12. As a measure of instantly available energy, ATP level declines about 80% from adult day 0 to day 12. Moreover, as an estimate of the potential for metabolic activity, the amount of superoxide anion declines about 95% from adult day 0 to day 12. Here we offer these assays service to monitor age-related changes in metabolic activity in kinds of factors, such as drugs and mutations.
The activity of specific enzymes has been shown to display age-related changes. As one of the enzymes, protein tyrosine kinase activity in worm extracts decreases about 60% from day 5 to day 15. We can also measure other enzymes that suit the particular needs of customers' requirements.
|Age-related morphology||Description||Method of measurement||Quantitative|
|Lipofuscin and advanced glycosylation end products||The spectral properties of lipofuscin and advanced glycosylation end products are characterized by an excitation maximum of about 340nm and an emission maximum of about 430nm, and resemble fluorescent spectra.||Spectrofluorimetry
-In vitro analysis of extracts
-In vivo with intact animals
Fluorescence/ electron microscopy
|DNA damage, protein carbonylation, yolk protein in the body cavity||DNA damage, by measuring the number of single-stand breaks and the levels of 5-methylcytosine.
Protein carbonylation, by determining a direct reaction of protein carbonyls with fluorescein thiosemicarbazide.
Yolk protein, by detecting the amount of accumulation in body cavity.
|DNA damage, protein carbonylation, measured by biochemical assays of extracts,
Yolk protein, measured by nomarski/electron microscopy.
|Metabolic activity||Carbon dioxide generation, measured by gas respirometry.
Oxygen consumption, measured by Clark type electrodes.
Heat production, measured by microcalorimetry.
ATP levels, measured by enzyme reactions that require ATP.
The amount of superoxide anion, measured by lucigenin-mediated luminescence.
|Various assays||DNA damage, protein carbonylation- yes
Yolk protein- no
|Enzymatic activities||Measurement of protein tyrosine kinase activity, and other enzymatic activities according to customers‘ requirements.||Biochemical assay||Yes|
* For research use only.
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