Exposure of zebrafish larvae to 3g and 5g hyper gravity between 5-6 days post-fertilization.

The general objective of the study was to determine modulation of gene expression by environmental factors, with a special emphasis on bone formation. For this reason, the specific period of treatment was chosen between 5-6 days post-fertilization (dpf), when bone formation and calcification are taking place. Zebrafish larvae were placed at 5 dpf into a Large Diameter Centrifuge and brought to a gravitational force of 3g or 5g for 24 hours. We show that this treatment causes a clear increase of bone formation, as illustrated by cranial skeleton staining of the bone matrix by Alizarin Red, by morphometric analysis of the resulting images and by gene expression studies of selected genes. Thus, a whole genome micro-array experiment was conducted to identify genes that may be involved in the observed effect on bone formation.

本研究的总体目标为探明环境因素对基因表达的调控作用，重点聚焦于骨形成过程。为此，我们选取受精后5~6天（days post-fertilization, dpf）这一特定处理阶段，此时斑马鱼幼体正处于骨形成与钙化的活跃阶段。实验中将受精后5天的斑马鱼幼体置于大直径离心机（Large Diameter Centrifuge）中，施加3g或5g的重力加速度，持续处理24小时。研究显示，该处理可显著促进骨形成，这一结果可通过以下方式验证：采用茜素红（Alizarin Red）对骨基质进行颅部骨骼染色、对所得染色图像开展形态计量分析，以及对选定基因进行基因表达检测。据此，我们开展了全基因组微阵列（whole genome micro-array）实验，以筛选可能参与上述骨形成调控效应的基因。