Triazole fungicides induce adipogenesis and repress osteoblastogenesis in zebrafish.

Triazoles are a major group of azole fungicides commonly used in agriculture, and veterinary and human medicine. Maternal exposure to certain triazole antifungal medication causes congenital malformations, including skeletal malformations. We hypothesized that triazoles used as pesticides in agriculture also pose a risk of causing skeletal malformations in developing embryos. In this study, teratogenic effects of three commonly used triazoles, cyproconazole, paclobutrazol, and triadimenol, were investigated in zebrafish, Danio rerio. Exposure to the triazole fungicides caused bone and cartilage malformations in developing zebrafish larvae. Data from whole-embryo transcriptomics with cyproconazole suggested that exposure to this compound induces adipogenesis while repressing skeletal development. Confirming this finding, the expression of selected bone and cartilage marker genes were significantly downregulated with triazoles exposure as determined by quantitative PCR. The expression of selected adipogenic genes was upregulated by the triazoles. Furthermore, exposure to each of the three triazoles induced adipogenesis and lipid droplet formation in vitro in 3T3-L1 pre-adipocyte cells. In vivo in zebrafish larvae, cyproconazole exposure caused lipid accumulation. These results suggest that exposure to triazoles promotes adipogenesis at the expense of skeletal development, and thus they expand the chemical group of bona fide bone to fat switchers.

A Layered Mounting Method for Extended Time-Lapse Confocal Microscopy of Whole Zebrafish Embryos.

Dynamics of development can be followed by confocal time-lapse microscopy of live transgenic zebrafish embryos expressing fluorescence in specific tissues or cells. A difficulty with imaging whole embryo development is that zebrafish embryos grow substantially in length. When mounted as regularly done in 0.3-1% low melt agarose, the agarose imposes growth restriction, leading to distortions in the soft embryo body. Yet, to perform confocal time-lapse microscopy, the embryo must be immobilized. This article describes a layered mounting method for zebrafish embryos that restrict the motility of the embryos while allowing for the unrestricted growth. The mounting is performed in layers of agarose at different concentrations. To demonstrate the usability of this method, whole embryo vascular, neuronal and muscle development was imaged in transgenic fish for 55 consecutive hours. This mounting method can be used for easy, low-cost imaging of whole zebrafish embryos using inverted microscopes without requirements of molds or special equipment.

Rigid-motion-invariant classification of 3-D textures.

This paper studies the problem of 3-D rigid-motion-invariant texture discrimination for discrete 3-D textures that are spatially homogeneous by modeling them as stationary Gaussian random fields. The latter property and our formulation of a 3-D rigid motion of a texture reduce the problem to the study of 3-D rotations of discrete textures. We formally develop the concept of 3-D texture rotations in the 3-D digital domain. We use this novel concept to define a "distance" between 3-D textures that remains invariant under all 3-D rigid motions of the texture. This concept of "distance" can be used for a monoscale or a multiscale 3-D rigid-motion-invariant testing of the statistical similarity of the 3-D textures. To compute the "distance" between any two rotations R(1) and R(2) of two given 3-D textures, we use the Kullback-Leibler divergence between 3-D Gaussian Markov random fields fitted to the rotated texture data. Then, the 3-D rigid-motion-invariant texture distance is the integral average, with respect to the Haar measure of the group SO(3), of all of these divergences when rotations R(1) and R(2) vary throughout SO(3). We also present an algorithm enabling the computation of the proposed 3-D rigid-motion-invariant texture distance as well as rules for 3-D rigid-motion-invariant texture discrimination/classification and experimental results demonstrating the capabilities of the proposed 3-D rigid-motion texture discrimination rules when applied in a multiscale setting, even on very general 3-D texture models.