Padronização da técnica de qPCR para quantificação do comprimento relativo telomérico no Caenorhabditis elegans

Abstract

Telomeres have a fundamental role in cellular replication and through their length it is possible to estimate cellular life span mean. Evidence has shown that factors such as environmental, social, nutritional and physiopathological may be implicated in telomere length variation. It has been demonstrated that accelerated telomere shortening may be related to the early onset of several healthcare problems related to aging. Once determined the importance of telomere length analyses, this study aimed to standardize a qPCR based technique to quantify relative telomere length in Caenorhabditis elegans. The use of this model has already shown to be viable in telomere-based studies. Studies show that due to its main characteristics, such as, sort life cycle, easy manipulation, genetic versatility and telomere complex similar to mammals, this model may be considered ideal to investigate initial molecular mechanisms that may interfere in telomere length. Such characteristics are, especially, valid for the developing of upcoming researches based on and directed to vertebrate models, aiming at an effective future clinical application. It was also taken in consideration the interest in measuring C. elegans telomere length in a less laborer but more accessible and faster manner, overcoming the limits of less efficient techniques standardized for this model. Strains N2 (Wild Type), CB3192 (Wild type), with a known longer telomere length, YA1059 strain, which has been previously shown to have a shorter telomere. Our results demonstrated linear coefficients for each gene with values proximate to 1(Telomere R² = 0,95 / C44B7.8 - R² = 0,91 / T09F3.3 - R² = 0,91 / F25B4.6 - R² = 0,93) and significant differences between strains with distinct telomere size (p< 0,0001 for CB3192 > N2 e CB3192 > YA1059; p= 0,0069 for N2 > YA1059). Results found, confirm the efficiency of the technique in relative telomere length measuring in C. elegans and its sensibility towards different strains. However the need of validating the qPCR technique through a gold standard method is still necessary.