Differential expression of dopaminergic cell markers in the adult zebrafish forebrain

Although the simultaneous presence of tyrosine hydroxylase (TH), aromatic amino acid decarboxylase (AADC), dopamine transporter (DAT), and vesicular monoamine transporter 2 (VMAT2) is considered as a phenotypic signature of dopamine (DA) neurons, it has been suggested that they are not uniformly expressed in all dopaminergic brain nuclei. Moreover, in nonmammalian vertebrates, two tyrosine hydroxylase genes (TH1 and TH2) are found, and they exhibit different expression patterns in zebrafish brains. Here we present a detailed description of the distribution of TH1, TH2, AADC, DAT, and VMAT2 transcripts, in relation to TH and DA immunoreactivity to better characterize dopaminergic nuclei in the adult zebrafish forebrain. TH2-positive cells in the hypothalamus are strongly DA immunoreactive (DAir), providing direct evidence that they are dopaminergic. DAir cells are also found in most TH1-positive or TH-immunoreactive (THir) nuclei. However, the DAir signal was weaker than THir in the olfactory bulb, telencephalon, ventral thalamus, pretectum, and some posterior tubercular and preoptic nuclei. These cell populations also exhibited low levels of VMAT2 transcripts, suggesting that low DA is due to a lower vesicular DA accumulation. In contrast, cell populations with low levels of AADC did not always have low levels of DA. DAT transcripts were abundantly expressed in most of the TH1- or TH2-positive territories. In addition, DAT and/or VMAT2 transcripts were found in some periventricular cell populations such as in the telencephalon without TH1 or TH2 expression. Thus, expression patterns of dopaminergic cell markers are not homogeneous, suggesting that the gene regulatory logic determining the dopaminergic phenotype is unexpectedly complex.