Abstract—

The embryonic and larval stages of amphibian development are promising model objects for developmental biology, experimental biology, and medicine. However, the existing methods of laboratory research do not take into account the biological features of model amphibian species. In addition, not enough attention is paid to standardizing the experimental conditions and minimizing the experimenter’s influence on their results. The fact that the experimental research methodology developed for one amphibian species cannot be used for similar research on another species without significant modification with consideration of the biology of this object is also not accounted for. The article summarizes the results of many years of research devoted to the study of factors that influence the results of laboratory experiments on the embryonic developmental stages of the common toad Bufo bufo. By the example of this species, new methodological approaches to laboratory research have been developed, allowing consideration for the species’ particular biological features, minimization in the heterogeneity of the effects from previously uncontrolled factors, and standardizing the conditions for conducting experiments. Maintaining the temperature at 15–17°C throughout the experiment, taking regular measurements at regular intervals (8 h), and conducting each test at two values of the initial egg density (recommended 30 and 120 eggs in a single string per standard aquarium) is recommend. When setting up experiments, one should take into account the influence of not only “bulk” but also “surface” and “linear” densities. It is also shown that any additional effects of the experimenter on the embryos associated with the removal of dead individuals performed during the experiments can have a significant impact on the result; therefore, such effects should be minimized.

中文翻译：

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蟾蜍胚胎发育的实验研究：方法论方面

摘要-

两栖动物发育的胚胎和幼虫阶段是发展生物学，实验生物学和医学的有希望的模型对象。但是，现有的实验室研究方法并未考虑模型两栖动物的生物学特征。另外，对标准化实验条件和使实验者对其结果的影响最小化没有给予足够的重视。对于一个两栖动物物种开发的实验研究方法不能在不考虑该对象生物学的情况下进行重大修改的情况下不能用于另一物种的类似研究，这一事实也没有得到说明。蟾蜍。以该物种为例，已经开发了用于实验室研究的新方法论方法，可以考虑该物种的特殊生物学特征，将以前不受控制的因素造成的影响的异质性降至最低，并标准化进行实验的条件。在整个实验过程中，将温度保持在15–17°C，以固定的时间间隔（8 h）进行定期测量，并以两个初始卵密度值（每个标准水族箱建议在单个字符串中推荐30和120个卵）进行每次测试建议。进行实验时，不仅应考虑“体”密度的影响，还应考虑“表面”和“线性”密度的影响。还表明，在实验过程中，实验者对胚胎的任何其他影响（与去除死者的行为有关）都可能对结果产生重大影响。因此，应尽量减少这种影响。