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Despite being considered a model organism in toxicity studies, particularly in assessing the environmental impact of endocrine disrupting compounds (EDCs) and other chemicals, the molecular basis of development is largely unknown in  Chironomus riparius . We have characterized the expression patterns of important genes involved in the ecdysone pathway from embryos to pupa, but specially during the different phases of  C .  riparius  fourth larval instar, according to the development of genital and thoracic imaginal discs. Real-Time PCR was used to analyze:  EcR  and  usp , two genes encoding the two dimerizing partners of the functional ecdysone receptor;  E74 , an early response gene induced by ecdysteroids;  vg  (vitellogenin), an effector gene;  hsp70  and  hsc70 , two heat-shock genes involved in the correct folding of the ecdysone receptor; and  rpL13 , as a part of the ribosomal machinery. Our results show for the first time stage and sex-dependent variations in ecdysone-responsive genes, specially during the late larval stage of  C .  riparius . The induction in the expression of  EcR  and  usp  during the VII-VIII phase of the fourth instar is concomitant with a coordinated response in the activity of the other genes analyzed, suggesting the moment where larvae prepare for pupation. This work is particularly relevant given that most of the analyzed genes have been proposed previously in this species as sensitive biomarkers for the toxicological evaluation of aquatic ecosystems. Identifying the natural regulation of these molecular endpoints throughout the  Chironomus  development will contribute to a more in-depth and accurate evaluation of the disrupting effects of EDCs in ecotoxicological studies.

Ecdysone-Related Biomarkers of Toxicity in the Model Organism Chironomus riparius: Stage and Sex-Dependent Variations in Gene Expression Profiles