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A Rapid, Non-invasive Method for Anatomical Observations of Tadpole Vertebrae in Vivo

Abstract

The tadpole is a critical stage in the amphibian life cycle and plays an important role during the transition from the aquatic to the terrestrial stage. However, there is a large gap in tadpole research, which represents a vital component of our understanding of the diversity and complexity of the life history traits of amphibians, especially their developmental biology. Some aspects of this gap are due to limited research approaches. To date, X-ray microcomputed tomography (micro-CT) has been widely used to conduct osteology research in adult amphibians and reptiles, but little is known about whether this tool can be applied in tadpole studies. Thus, we compared the results of two methods (the bone-cartilage double-staining technique and micro-CT) to study vertebrae in tadpole specimens. The results revealed no significant difference between the two methods in determining the number of vertebrae, and micro-CT represents a rapid, non-invasive, reliable method of studying tadpole vertebrae. When scanning tadpoles, voltage is the most critical of the scanning parameters (voltage, current and scan time), and moderate scanning parameters are recommended. In addition, micro-CT performed better using specimens stored in 70% ethanol than those preserved in 10% formalin. Finally, we suggest that micro-CT should be more widely applied in herpetological research to increase specimen utilization

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