Pyriproxyfen is an agricultural chemical pesticide that has been detected in the aquatic environment. This study aimed to clarify the effects of pyriproxyfen on the growth as well as thyroid hormone- and growth-related gene expression of zebrafish (Danio rerio) during its early life stage. Pyriproxyfen exhibited a lethal effect in a concentration-dependent manner: the lowest and no effect concentrations were 250.7 and 111.7 μg/L, respectively. These concentrations were considerably higher than the residual environmental concentrations, indicating the low risk of this pesticide when present at such concentrations. In the zebrafish group treated with 56.6 μg/L pyriproxyfen, thyroid hormone receptor β gene expression levels remained unchanged, whereas thyroid-stimulating hormone β subunit, iodtyronin deiodinase 2, and thyroid hormone receptor α gene expression levels significantly decreased compared with the control group expression levels. In zebrafish treated with 111.7 or 250.7 μg/L pyriproxyfen, iodtyronin deiodinase 1 gene expression levels significantly increased. These results indicate that pyriproxyfen disrupts thyroid hormone activity in zebrafish. Furthermore, pyriproxyfen exposure inhibited zebrafish growth; consequently, we examined the expression of growth hormone (gh) and insulin-like growth factor-I (igf-1), which are important for growth. Pyriproxyfen exposure suppressed gh expression; however, the igf-1 expression levels remained unchanged. Therefore, growth inhibition due to pyriproxyfen exposure was attributed to the suppression of gh expression.
Phthalate and non-phthalate plasticizers are used in polymer materials, such as plastic and rubber. It has recently been found that diisobutyl adipate (DIBA), which is considered an environmentally safe non-phthalate plasticizer, potentially acts as a thyroid disruptor in fish. Here, we investigated the sexual hormone effects of DIBA based on the expression levels of genes that respond to endocrine disruption and sexual hormone activity in the livers and gonads, and on gonadal sexual differentiation in Japanese medaka. Compared with the control group, the mRNA expression of chgH, vtg1, vtg2, and esr1 was significantly suppressed in the livers of DIBA exposed XX individuals. Furthermore, the mRNA expression of gsdf was significantly upregulated and downregulated in the gonads of XX and XY individuals, respectively. The mRNA expressions of esr1 and esr2b were significantly suppressed by DIBA exposure in the gonads of both XX and XY individuals. These observations suggest that DIBA has potential androgenic activity in Japanese medaka. However, normal testes and ovaries were observed in respective XY and XX medaka after DIBA exposure; therefore, these results suggest that DIBA may have weak androgenic activity.
Cathelicidin is a family of antimicrobial peptides in vertebrates that plays an important role in resistance and immunization against pathogenic microorganisms. In the present study, the full-length cDNA sequences of four novel cathelicidins (cathelicidin-1 to cathelicidin-4) in the tiger frog Hoplobatrachus rugulosus, encoding 153, 188, 132, and 160 amino acids, respectively, were firstly cloned by rapid amplification of the cDNA ends (RACE) technique. Sequence comparison and phylogenetic tree analysis indicated that the structures of the four cathelicidins are highly diverse. Afterwards, the tissue distribution profiles and antimicrobial patterns of cathelicidins in H. rugulosus were determined by real-time PCR. The four cathelicidins showed tissue-specific distribution patterns in the healthy frogs, and the transcriptional levels of cathelicidins exhibited a tissue- and time-dependency profile in the frogs challenged with pathogenic bacteria Aeromonas hydrophila for 72 h. The synthetic peptides of cathelicidin-1 and cathelicidin-2 exhibited broad-spectrum in vitro antimicrobial activity, and cathelicidins exerted antimicrobial activities through excessive induction of reactive oxygen species and direct disruption of the microbial membrane structure. In addition, the intraperitoneal injection of cathelicidin proteins significantly increased the marine medaka Oryzias melastigma resistance to bacterial challenges. The existence of multiple cathelicidins, their distinct tissue distribution patterns, and the inducible expression profiles suggest a sophisticated, highly redundant, and multilevel network of antimicrobial defense mechanisms in tiger frogs. This study provides evidence that cathelicidins have antimicrobial and immunomodulatory activities, and cathelicidins derived from H. rugulosus have potential therapeutic applications against pathogenic infections in aquaculture.
High molecular weight proteins are present abundantly in viperid venoms. The amino acid sequence can be highly variable, contributing to the structure and function diversity of snake venom protein. However, this variability remains poorly understood in many species. The study investigated the venom protein variability in a distinct clade of Asian pit vipers (Trimeresurus species complex) through comparative proteomics, applying gel electrophoresis (SDS-PAGE), liquid chromatography-tandem mass spectrometry (LCMS/MS), and bioinformatic approaches. The proteomes revealed a number of conserved protein families, within each are variably expressed protein paralogs that are unrelated to the snake phylogeny and geographic origin. The expression levels of two major enzymes, i.e., snake venom serine proteinase and metalloproteinase, correlate weakly with procoagulant and hemorrhagic activities, implying co-expression of other functionally versatile toxins in the venom. The phospholipase A2 (PLA2) abundance correlates strongly with its enzymatic activity, and a unique phenotype was discovered in two species expressing extremely little PLA2. The commercial mono-specific antivenom effectively neutralized the venoms' procoagulant and hemorrhagic effects but failed to inhibit the PLA2 activities. Instead, the PLA2 activities of all venoms were effectively inhibited by the small molecule inhibitor varespladib, suggesting its potential to be repurposed as a highly potent adjuvant therapeutic in snakebite envenoming.