Investigating Neotenic and Metamorphic Axolotl Brain Complexity: A Stereological and Immunohistochemical Perspective

The ability of certain tetrapods, such as amphibians, to regenerate complex structures, such as organs or limbs, is well-established, though this capacity varies significantly across species, with humans exhibiting limited regenerative potential. Ependymoglia cells in the ventricular region of the brain are known to exhibit proliferative properties during homeostasis and damage and to perform stem cell functions. This study investigated changes occurring in neurons and glia in the central nervous system following metamorphosis in axolotls. Morphological alterations in brain tissue, newly formed neurons, and cellular organizations in different brain regions were assessed using stereological and immunohistochemical methods, as well as light and electron microscopy. Interestingly, we observe no statistically significant difference in total neuron numbers in the telencephalon region between neotenic and metamorphic axolotls. However, the proliferation index and the numbers of cells expressing NeuN were significantly higher in metamorphic axolotls. Furthermore, structural changes in neuronal nuclei and myelin sheath organization were determined at the light and electron microscopic levels post-metamorphosis. Ultrastructural analyses revealed a change in chromatin organization from euchromatic to heterochromatic in neurons after metamorphosis, and morphological changes were also demonstrated in myelinated nerve fibers in the telencephalon. Additionally, mucopolysaccharide-containing secretory sacs were also identified on the apical surfaces of a subgroup of ependymoglia cells located in the lateral ventricle wall. Overall, this study sheds useful light on the intricate changes occurring in the central nervous system during metamorphosis in axolotls and provides valuable insights into the mechanisms underlying these processes.