The distribution of dopamine (DA)-containing cell bodies, fibers, and terminals in the brain and spinal cord of Lampetra fluviatilis was investigated by immunohistochemical means. In order to distinguish dopaminergic neurons from those using other catecholamines as the primary neurotransmitter, the distribution of dopamine-immunoreactive structures was compared to that of cell bodies, fibers, and terminals labelled with antibodies directed against the enzymes tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC), dopamine beta-hydroxylase (DBH), and phenylethanolamine-N-methyl transferase (PNMT). We define dopaminergic neurons as those that are simultaneously DA, TH, and AADC immunoreactive and at the same time DBH and PNMT nonreactive. The overall concentrations of dopamine, noradrenaline, and adrenaline and some of their metabolites were also measured via high-performance liquid chromatography of whole-brain extracts. Our results revealed the presence of 10 populations of dopaminergic neurons in the brain of the lamprey in the olfactory bulb, preoptic area, hypothalamus, rhombencephalon, and spinal cord. In addition, uniquely DA-immunoreactive neurons, in contact with the cerebrospinal fluid, were observed in the hypothalamus and spinal cord. Chromatography indicated that dopamine exists in considerably higher concentrations than noradrenaline in the lamprey brain, whereas adrenaline is absent, the latter finding being supported by our failure to observe any PNMT-immunoreactive cell bodies, fibers, or terminals. The dopaminergic system of the lamprey appears to share many features not only with that of other anamniotes but also with that of amniotes; however, as in teleosts, dopaminergic neurons in the midbrain corresponding to the substantia nigra, the retrorubral area, and the ventral tegmental area of other species do not exist in the lamprey.
The experiments reported here showed that application of serotonin (5-hydroxytryptamine, 5-HT) (100 microM) did not induce any significant current through the membranes of any of the spinal neurons studied (n = 62). At the same time, the membranes of most motoneurons and interneurons (15 of 18) underwent slight depolarization (2-6 mV) in the presence of 5-HT, which was not accompanied by any change in the input resistance of the cells. Depolarization to 10-20 mV occurred in some cells (3 of 18) of these functional groups, this being accompanied by 20-60% decreases in input resistance. The same concentration of 5-HT induced transient low-amplitude depolarization of most sensory spinal neurons (dorsal sensory cells), this changing smoothly to long-term hyperpolarization by 2-7 mV. The input resistance of the cell membranes in these cases showed no significant change (n = 8). Data were obtained which provided a better understanding of the mechanism by which 5-HT modulates the activity of spinal neurons. Thus, 5-HT facilitates chemoreceptive currents induced by application of NMDA to motoneurons and interneurons, while the NMDA responses of dorsal sensory cells were decreased by 5-HT. 5-HT affected the post-spike afterresponses of neurons. 5-HT significantly decreased the amplitude of afterhyperpolarization arising at the end of the descending phase of action potentials in motoneurons and interneurons and increased the amplitude of afterdepolarization in these types of cells. In sensory spinal neurons, 5-HT had no great effect on post-spike afterresponses. The results obtained here support the suggestion that 5-HT significantly modulates the activity of spinal neurons of different functional types. 5-HT facilitates excitation induced by subthreshold depolarization in motoneurons and some interneurons, facilitating the generation of rhythmic discharges by decreasing afterhyperpolarization. In sensory cells, 5-HT enhances inhibition due to hyperpolarization, suppressing NMDA currents. The differences in the effects of 5-HT on functionally diverse neurons are presumed to be associated with the combination of different types of 5-HT receptors on the membranes of these types of spinal neurons.
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