Determination and Occurrence of Retinoids in a Eutrophic Lake (Taihu Lake, China): Cyanobacteria Blooms Produce Teratogenic Retinal

Besides retinoic acids (RAs), some retinoids such as retinal (RAL) and retinol (ROH), which are considered as RA precursors in vertebrates, are also reported to be teratogenic agents. In this study we investigated four RA precursors including RAL, ROH, retinyl palmitate, and β-carotene in the eutrophic Taihu Lake, China, by developing a sensitive analytical method. RAL and β-carotene were widely detected in natural cyanobacteria blooms and lake water. Intracellular concentrations of RAL and β-carotene in blooms were 9.4 to 6.9 × 10³ and 3.4 to 1.8 × 10⁵ ng L^–1, respectively, and their concentrations in lake water were up to 1.4 × 10 ng L^–1 (RAL) and 9.8 × 10² ng L^–1 (β-carotene). The good correlation between intracellular concentrations of RAL and RAs implied that RAL was involved in the production of RAs by cyanobacteria blooms. Further examination of 39 cyanobacteria and algae species revealed that most species could produce RAL and β-carotene. The greatest amount of RAL was found in <i>Chlamydomonas</i> sp. (FACHB-715; 1.9 × 10³ ng g^–1 dry weight). As the main cyanobacteria in Taihu Lake, many <i>Microcystis</i> species could produce high amounts of RAL and were thought to greatly contribute to the production of RAL measured in the blooms. Productions of RAL and β-carotene by cyanobacteria were associated with species, origin location, and growth stage. The results in this study present the existence of a potential risk to aquatic animals living in a eutrophic environment from a high concentration of RAL in cyanobacteria blooms and also provide a clue for further investigating the mechanism underlying the biosynthetic pathway of RAs in cyanobacteria and algae

Biosensor Medaka for Monitoring Intersex Caused by Estrogenic Chemicals

Estrogenic chemicals can induce intersex in fish species leading to disturbance of spermatogenesis and impairment of reproductive success. To overcome the shortcomings of conventional histopathological observation on intersex (low sensitivity, relatively poor accuracy, long experimental periods, as well as laborious and time-consuming), we generated a pMOSP1-EGFP transgenic medaka fish model. In this transgenic fish, the green fluorescence protein (GFP) reporter gene was derived by the regulatory elements of the OSP1 gene, which is a specific and sensitive molecular biomarker for indicating intersex occurrence in male medaka fish exposed to estrogenic chemicals. The transgenic GFP was faithfully expressed in ovaries and in testes with intersex, perfectly mimicking the expression pattern of endogenous OSP1. In intersex testis, the diameters of primary oocytes which could be distinguished by GFP fluorescence observation were as small as 10 μm, lower than that (more than 20 μm) which is observable by histopathology. Using the novel transgenic medaka fish, intersex was observed after 90-day exposure to 0.75 ng/L 17α-ethinyloestradiol (EE₂) (0–90 dph), but only at concentrations of at least 1.38 ng/L EE₂ by histopathology. An effectiveness of a short-term in vivo assay for screening estrogenic chemicals that can monitor intersex appearance at early sex developmental stage (about 30 dph) in male medaka fish was also demonstrated by assessing the intersex induction of EE₂, 17β-estradiol and 4-nonylphenol. This newly developed assay provides an enhanced ability for screening and testing estrogenic chemicals with the potential to induce intersex and studying their biological impacts