Fish bioaccumulation and biomarkers in environmental risk assessment: a review

The in situ hybridization (ISH) technique allows the sites of expression of particular genes to be detected. This protocol describes ISH of digoxigenin-labeled antisense RNA probes to whole-mount zebrafish embryos. In our method, PCR-amplified sequence of a gene of interest is used as a template for the synthesis of an antisense RNA probe, which is labeled with digoxigenin-linked nucleotides. Embryos are fixed and permeabilized before being soaked in the digoxigenin-labeled probe. We use conditions that favor specific hybridization to complementary mRNA sequences in the tissue(s) expressing the corresponding gene. After washing away excess probe, hybrids are detected by immunohistochemistry using an alkaline phosphatase-conjugated antibody against digoxigenin and a chromogenic substrate. The whole procedure takes only 3 days and, because ISH conditions are the same for each probe tested, allows high throughput analysis of zebrafish gene expression during embryogenesis.

High-resolution in situ hybridization to whole-mount zebrafish embryos

The role of antioxidants in the chemistry of oxidative stress: A review.

Reactions of chlorine with inorganic and organic compounds during water treatment—Kinetics and mechanisms: A critical review

https://recipp.ipp.pt/bitstream/10400.22/3012/4/ART_LuciaSantos_2010_GRAQ.pdf

Ecotoxicological aspects related to the presence of pharmaceuticals in the aquatic environment

Ethinyloestradiol — an undesired fish contraceptive?

In zebrafish (Danio rerio) pigmentation is initiated during embryogenesis and begins in the retinal epithelium and in the melanophores. The pigment cells develop rapidly, and within hours they constitute a prominent feature of the embryo. In order to improve signal detection by whole mount in situ hybridization, confocal microscopy, or expression of GFP, embryos may be treated with 1-phenyl 2-thiourea (PTU) during embryogenesis. PTU inhibits melanogenesis by blocking all tyrosinase-dependent steps in the melanin pathway but can be toxic at high concentrations. The embryos remain transparent as long as the PTU treatment is continued. However, PTU treatment must be initiated before the initial pigmentation because it does not remove already formed pigment. Here we provide a protocol for generating transparent zebrafish while avoiding the toxic and teratogenic effects of PTU treatment.

Generating transparent zebrafish: a refined method to improve detection of gene expression during embryonic development.

Coronavirus disease 2019 (COVID-19) has shown high infection and mortality rates all over the world, and despite the global efforts, there is so far no specific therapy available for COVID-19. Interestingly, while the severity and mortality of COVID-19 are higher in males than in females, the underlying molecular mechanisms are unclear. In this review, we explore sex-related differences that may be contributing factors to the observed male-biased mortality from COVID-19. Males are considered the weaker sex in aspects related to endurance and infection control. Studies show that viral RNA clearance is delayed in males with COVID-19. A recent study has indicated that the testis can harbor coronavirus, and consequently, males show delayed viral clearance. However, the role of testis involvement in COVID-19 severity and mortality needs further research. Males and females show a distinct difference in immune system responses with females eliciting stronger immune responses to pathogens. This difference in immune system responses may be a major contributing factor to viral load, disease severity, and mortality. In addition, differences in sex hormone milieus could also be a determinant of viral infections as estrogen has immunoenhancing effects while testosterone has immunosuppressive effects. The sex-specific severity of COVID-19 infections indicates that further research on understanding the sex differences is needed. Inclusion of both males and females in basic research and clinical trials is required to provide critical information on sex-related differences that may help to better understand disease outcome and therapy.

/pdf/sex-differences-in-severity-and-mortality-from-covid-19-are-2irh92ydx5.pdf

Sex differences in severity and mortality from COVID-19: are males more vulnerable?

Sex determination is the process deciding the sex of a developing embryo. This is usually determined genetically; however it is a delicate process, which in many cases can be influenced by environmental factors. The mechanisms controlling zebrafish sex determination and differentiation are not known. To date no sex linked genes have been identified in zebrafish and no sex chromosomes have been identified. However, a number of genes, as presented here, have been linked to the process of sex determination or differentiation in zebrafish. The zebrafish FTZ-F1 genes are of central interest as they are involved in regulating interrenal development and thereby steroid biosynthesis, as well as that they show expression patterns congruent with reproductive tissue differentiation and function. Zebrafish can be sex reversed by exposure to estrogens, suggesting that the estrogen levels are crucial during sex differentiation. The Cyp19 gene product aromatase converts testosterone into 17 beta-estradiol, and when inhibited leads to male to female sex reversal. FTZ-F1 genes are strongly linked to steroid biosynthesis and the regulatory region of Cyp19 contains binding sites for FTZ-F1 genes, further linking FTZ-F1 to this process. The role of FTZ-F1 and other candidates for zebrafish sex determination and differentiation is in focus of this review.

/pdf/zebrafish-sex-determination-and-differentiation-involvement-3krf675xcu.pdf

Zebrafish sex determination and differentiation: involvement of FTZ-F1 genes.

As a result of mining, forestry, waste disposal and fuel combustion, our environment is becoming increasingly contaminated with heavy metals The aquatic environment receives waste products from such activities and may be the final depository for these anthropogenically remobilized heavy metals In order to understand the impact of heavy metals on aquatic biota it is important to characterize the mechanisms available for aquatic life to transport, immobilize and excrete heavy metals

Per-Erik Olsson

Papers

Ethinyloestradiol — an undesired fish contraceptive?

Generating transparent zebrafish: a refined method to improve detection of gene expression during embryonic development.

Sex differences in severity and mortality from COVID-19: are males more vulnerable?

Zebrafish sex determination and differentiation: involvement of FTZ-F1 genes.

Mechanisms of heavy metal accumulation and toxicity in fish