Effects of MP Polyethylene Microparticles on Microbiome and Inflammatory Response of Larval Zebrafish.

Plastic polymers have quickly become one of the most abundant materials on Earth due to their low production cost and high versatility. Unfortunately, some of the discarded plastic can make its way into the environment and become fragmented into smaller microscopic particles, termed secondary microplastics (MP). In addition, primary MP, purposely manufactured microscopic plastic particles, can also make their way into our environment via various routes. Owing to their size and resilience, these MP can then be easily ingested by living organisms. The effect of MP particles on living organisms is suspected to have negative implications, especially during early development. In this study, we examined the effects of polyethylene MP ingestion for four and ten days of exposure starting at 5 days post-fertilization (dpf). In particular, we examined the effects of polyethylene MP exposure on resting metabolic rate, on gene expression of several inflammatory and oxidative stress linked genes, and on microbiome composition between treatments. Overall, we found no evidence of broad metabolic disturbances or inflammatory markers in MP-exposed fish for either period of time. However, there was a significant increase in the oxidative stress mediator L-FABP that occurred at 15 dpf. Furthermore, the microbiome was disrupted by MP exposure, with evidence of an increased abundance of Bacteroidetes in MP fish, a combination frequently found in intestinal pathologies. Thus, it appears that acute polyethylene MP exposure can increase oxidative stress and dysbiosis, which may render the animal more susceptible to diseases.

Metabolic regulation by the PGC-1α and PGC-1ß coactivators in larval zebrafish (Danio rerio).

The peroxisome proliferator activated receptor γ coactivator-1 (PGC-1) family is composed of three coactivators whose role in regulating mammalian bioenergetics regulation is clear, but is much less certain in other vertebrates. Current evidence suggests that in fish, PGC-1α and PGC-1ß may exhibit much less redundancy in the control of fatty acid oxidation and mitochondrial biogenesis compared to mammals. To assess these roles directly, we knocked down PGC-1α and PGC-1ß expression with morpholinos in zebrafish embryos, and we investigated the resulting molecular and physiological phenotypes. First, we found no effects of either morpholinos on larval hatching, heart rates and oxygen consumption over the first few days of development. Next, at 3 days post fertilization (dpf), we confirmed by real time PCR a specific knock down of both coactivators, that resulted in a significant reduction in the transcript levels of citrate synthase (CS), 3-hydroxyacyl-CoA dehydrogenase (HOAD), and medium-chain acyl-coenzyme A dehydrogenase (MCAD) in both morphant groups. However, there was no effect on transcription factors' gene expression except for a marked reduction in estrogen related receptor α (ERRα) transcripts in PGC-1α morphants. Finally, we assessed whole embryonic enzyme activity for CS, cytochrome oxidase (COX), HOAD and carnitine palmitoyltransferase I (CPT-1) at 4 dpf. The only significant effect of the knockdown was a reduced CS activity in PGC-1α morphants and a counterintuitive increase of cytochrome oxidase activity in PGC-1ß morphants. Overall, our results indicate that in larval zebrafish, PGC-1α and PGC-1ß both play a role in regulating expression of important mitochondrial genes potentially through ERRα.

Prostaglandin E2 promotes embryonic vascular development and maturation in zebrafish.

Prostaglandin (PG)-E2 is essential for growth and development of vertebrates. PGE2 binds to G-coupled receptors to regulate embryonic stem cell differentiation and maintains tissue homeostasis. Overproduction of PGE2 by breast tumor cells promotes aggressive breast cancer phenotypes and tumor-associated lymphangiogenesis. In this study, we investigated novel roles of PGE2 in early embryonic vascular development and maturation with the microinjection of PGE2 in fertilized zebrafish (Danio rerio) eggs. We injected Texas Red dextran to trace vascular development. Embryos injected with the solvent of PGE2 served as vehicle. Distinct developmental changes were noted from 28-96 h post fertilization (hpf), showing an increase in embryonic tail flicks, pigmentation, growth, hatching and larval movement post-hatching in the PGE2-injected group compared to the vehicle. We recorded a significant increase in trunk vascular fluorescence and maturation of vascular anatomy, embryo heartbeat and blood vessel formation in the PGE2 injected group. At 96 hpf, all larvae were euthanized to measure vascular marker mRNA expression. We observed a significant increase in the expression of stem cell markers efnb2a, ephb4a, angiogenesis markers vegfa, kdrl, etv2 and lymphangiogenesis marker prox1 in the PGE2-group compared to the vehicle. This study shows the novel roles of PGE2 in promoting embryonic vascular maturation and angiogenesis in zebrafish.This article has an associated First Person interview with the first author of the paper.

Transcriptional effects of polyethylene microplastics ingestion in developing zebrafish (Danio rerio).

Over the last few decades, plastic waste has become an increasing environmental concern as it accumulates in every environment on our planet. Though traditionally seen as a macroscopic problem (i.e., large plastic debris), plastic pollution is also evident at smaller scales. Indeed, the intentional industrial production of small plastic particles and the physical degradation of larger plastic debris have overtime resulted in an increased environmental prevalence of smaller plastic particles, including microplastics. While the effects of these small polymers on marine biota have been an important research focus, recent global surveys indicate that our freshwater lakes and rivers are also plagued by microplastics. However, despite these discoveries we currently have a limited understanding of the impact these particles may have on freshwater animals, particularly on vertebrate species. Thus, the aim of the present study was to assess the impact of high concentrations of microplastics (5 and 20 mg.L-1) on the early life stages in zebrafish, a model freshwater vertebrate model. To do this, we exposed embryonic and larval zebrafish to fluorescently labelled polyethylene microspheres for up to 14 days and assessed their microplastic content, growth, hatching and oxygen consumption rates. We then explored the molecular underpinnings of the microplastic response by RNA sequencing. Over the course of the exposure, we observed a consistent accumulation of microplastics in the gastrointestinal tract of the fish in a concentration dependent manner, but could not detect any detrimental effects of these particles on larval development, growth or metabolism. However, whole animal transcriptomics revealed that microplastics induced a transient and extensive change in larval gene expression within 48 h exposure, which largely disappeared by 14 days. However, as these transcriptional changes occurred during a critical period of larval development, we suggest that an evaluation of the potential long-term impact of these particles is warranted.