Lipid dynamics in zebrafish embryonic development observed by DESI-MS imaging and nanoelectrospray-MS.

The zebrafish Danio rerio is a model vertebrate organism for understanding biological mechanisms. Recent studies have explored using zebrafish as a model for lipid-related diseases, for in vivo fish bioassays, and for embryonic toxicity experiments. Mass spectrometry (MS) and MS imaging are established tools for lipid profiling and spatial mapping of biomolecules and offer rapid, sensitive, and simple analytical protocols for zebrafish analysis. When ambient ionization techniques are used, ions are generated in native environmental conditions, requiring neither sample preparation nor separation of molecules prior to MS. We used two direct MS techniques to describe the dynamics of the lipid profile during zebrafish embryonic development from 0 to 96 hours post-fertilization and to explore these analytical approaches as molecular diagnostic assays. Desorption electrospray ionization (DESI) MS imaging followed by nanoelectrospray (nESI) MS and tandem MS (MS/MS) were used in positive and negative ion modes, allowing the detection of a large variety of phosphatidylglycerols, phosphatidylcholines, phosphatidylinositols, free fatty acids, triacylglycerols, ubiquinone, squalene, and other lipids, and revealed information on the spatial distributions of lipids within the embryo and on lipid molecular structure. Differences were observed in the relative ion abundances of free fatty acids, triacylglycerols, and ubiquinone - essentially localized to the yolk - across developmental stages, whereas no relevant differences were found in the distribution of complex membrane glycerophospholipids, indicating conserved lipid constitution. Embryos exposed to trichloroethylene for 72 hours exhibited an altered lipid profile, indicating the potential utility of this technique for testing the effects of environmental contaminants.

Effects of cypermethrin and carbofuran on certain hematological parameters and prediction of their recovery in a freshwater teleost, Labeo rohita (Hamilton).

Sublethal effects of cypermethrin and carbofuran on hematological parameters and their complete recovery were studied in Labeo rohita as a function of exposure time. Exposure of Labeo rohita to sublethal levels of cypermethrin and carbofuran resulted in significantly (P<0.05) lower values for erythrocyte count (RBC), hemoglobin content (Hb), and hematocrit compared with the control group. In contrast, there was a significant increase (P<0.05) in leukocyte count (TLC) in the pesticide-treated group. Mean cell volume (MCV) and mean cell hemoglobin (MCH) increased in response to both pesticides during the study. The rate of recovery or extrapolation time between Days 0 and 28 of the recovery period revealed that RBC completely recovered from cypermethrin toxicity on Days 80, 86.6, and 90 and TLC recovered on Days 60, 62.7, and 63.3 in fish exposed to 0.16, 0.40, and 0.80 microL/L cypermethrin, respectively. In the case of carbofuran toxicity, the complete recovery for RBC was on Days 100, 101.5, and 102, and for TLC, on Days 77, 77.6, and 79.3 in fish exposed to 0.06, 0.15, and 0.30 mg/L carbofuran, respectively. TLC recovered faster than RBC. RBC and TLC of fish exposed to carbofuran took more time to complete recovery than RBC and TLC in fish exposed to cypermethrin.