Immunolocalization of aromatic L-amino acid decarboxylase, tyrosine hydroxylase, dopamine, and serotonin in the forebrain of Ambystoma mexicanum.

To improve basic knowledge about the neurochemical organization of the urodele brain, and to study discrepancies in the localization of monoaminergic markers, we immunohistochemically charted the distribution of four such markers (tyrosine hydroxylase, aromatic L-amino acid decarboxylase, dopamine, and serotonin) in the axolotl (Ambystoma mexicanum) forebrain. Catecholaminergic and serotoninergic systems were found in similar locations to those seen in other Urodela. As seen in other vertebrates, the localization of the different monoaminergic markers reveals some inconsistencies. Cells that are exclusively tyrosine hydroxylase-immunoreactive are observed in the olfactory bulb, anterior olfactory nucleus/nucleus accumbens region, the epichiasmatic portion of the preoptic nucleus, and in the pars intercalaris thalami, whereas cells that are only labelled by aromatic L-amino acid decarboxylase are seen in the anterior olfactory nucleus/nucleus accumbens region, the bed nuclei of the anterior commissure, the posterior portion of the preoptic nucleus, the ventral hypothalamus, and the pars intercalaris thalami. The presence of cells solely serotonin (5-HT)-immunoreactive is suggested for the nucleus infundibularis dorsalis. Conversely, there were no areas that appeared to be exclusively immunoreactive for dopamine. Double-labelling for aromatic L-amino acid decarboxylase/tyrosine hydroxylase and aromatic L-amino acid decarboxylase/serotonin, together with cell counting, confirmed the existence of neurons that express only one monoaminergic marker in amphibian, supporting the hypothesis that these cells are universally present in the central nervous system of vertebrates.

Acrolein fixation for immunostaining tyrosine-hydroxylase in paraffin sections.

Routine histological fixatives barely preserve tyrosine-hydroxylase immunoreactivity in paraffin sections. fixation in 5% acrolein in phosphate buffer, pH 7.4, resulted in good preservation of the enzyme in the tissues investigated.

Immunohistochemical investigation of tyrosine-hydroxylase in the islets of Langerhans of adult mice, rats and guinea pigs.

As an indirect way to establish whether catecholamines are actually elaborated in the islets of Langerhans of adult albino and pigmented mice and guinea pigs, or albino rats, the presence of tyrosine-hydroxylase was immunohistochemically investigated using a highly specific polyclonal serum against the enzyme. The antiserum was applied to paraffin sections of pancreases fixed either in acrolein or Bouin's fluid. Such sections were then treated with the avidin-biotin method. Tyrosine-hydroxylase was found exclusively in the beta cells of rats and in those of the two strains of mice. Results in the guinea pig varied according to whether the animals were pigmented or albino. In pigmented specimens, the enzyme was detected in beta and non-beta cells, whereas in albino animals it was undetectable in any of the islet cells. All these observations were performed in material fixed in acrolein. Bouin's fluid resulted a rather poor fixative for the detection of the enzyme in the islets. The results are compared with those of other authors who investigated the presence of monoamines in the islets of Langerhans. Since tyrosine-hydroxylase is a specific marker for catecholamine-synthesizing cells, it is concluded that insular cells have the ability to elaborate these substances.