ABSTRACT
The effects of bisphenol A (BPA) on the endocrine system of vertebrates have been demonstrated in several studies. Here, we report the impact of BPA on the developmental progression and expression of heat shock protein genes on the terrestrial insect Sesamia nonagrioides (Lepidoptera: Noctuidae). S. nonagrioides 1st instar larvae were exposed until the end of 6th (last) instar to selected concentrations of BPA (1 µg/L, 10 µg/L, 100 µg/L, 1 mg/L and 10 mg/L) applied in their artificial diets. The lower doses of BPA (1-10 µg/L) were found to decrease larvae's weight while the 100 µg/L dose increased it. The higher doses of BPA were found to induce various abnormal phenotypes during 5th instar larval molting, larval-pupal transformation and metamorphosis. The developmental and metamorphosis endpoints presented here may indicate the possible impact of BPA on terrestrial insects. Additionally, 6th instar larvae were injected with several concentrations of BPA. Semi-quantitative and Real-Time PCR assays were used to identify the effects of BPA in the transcriptional regulation of five heat shock protein genes (SnoHsp19.5, SnoHsp20.8, SnoHsp70, SnoHsc70 and SnoHsp83). Application of BPA by feeding or by injection induced the synthesis of the SnoHsp19.5 and SnoHsp20.8 mRNAs. The expression levels of SnoHsp70 were not affected. In contrast, SnoHsc70 and SnoHsp83, which play a pivotal role in vertebrate sex steroid signal transduction, were elevated by BPA. Our results suggest that SnoHsp19.5, SnoHsp20.8, SnoHsp83 and SnoHsc70 genes can be modulated by BPA.
Subject(s)
Benzhydryl Compounds/pharmacology , Estrogens, Non-Steroidal/pharmacology , Heat-Shock Proteins/metabolism , Insect Proteins/metabolism , Moths/drug effects , Phenols/pharmacology , Animals , Benzhydryl Compounds/administration & dosage , Dose-Response Relationship, Drug , Estrogens, Non-Steroidal/administration & dosage , Gene Expression Regulation , Larva/drug effects , Larva/growth & development , Larva/metabolism , Moths/growth & development , Moths/metabolism , Phenols/administration & dosage , Pupa/drug effects , Pupa/growth & development , Pupa/metabolism , Real-Time Polymerase Chain ReactionABSTRACT
We obtained a full-length cDNA encoding a carboxylesterase in Sesamia nonagrioides. The complete cDNA sequence is comprised of 1838 bp with an open reading frame encoding 576 amino acid residues with predicted molecular mass of 64.24 kDa. The deduced amino acid sequence showed high identity to JHE-Related of Trichoplusia ni (65% amino acid identity) and 49-46% amino acid identity to JHEs of other lepidopterans and contained all five functional motifs of insect JHEs. The gene has been termed as SnJHE-Related (SnJHER) to denote its similarity to other insect JHE genes and the occurrence of an unusual cysteine residue immediately adjacent to the catalytic serine, instead of the conventional alanine residue. Phylogenetic analyses localised SnJHER together with TnJHER in a branch of the lepidopteran's JHEs group, with other carboxylesterases (COEs) occuring in separated groups. The JH analog methoprene did not affect the expression of SnJHER in contrast to other insect JHEs. Additionally, ecdysteroid analogs induced SnJHER gene expression. The SnJHER mRNA levels were higher in long-day non-diapausing larvae than in short-day diapausing ones. In the fifth instar of non-diapausing and ninth instar of diapausing larvae, the SnJHER mRNAs reached higher expression levels on the days close to each larval molt. In the last (sixth) non-diapausing larval instar, SnJHER mRNA levels peaked in the intermolt period but were lower than during the fifth instar.
