Elsevier

Neuroscience

Volume 127, Issue 1, 2004, Pages 147-154
Neuroscience

The too few mutant selectively affects subgroups of monoaminergic neurons in the zebrafish forebrain

https://doi.org/10.1016/j.neuroscience.2004.05.004Get rights and content

Abstract

Monoaminergic neurons are present in small numbers and in multiple distinct locations of the vertebrate CNS. They are involved in important functions such as movement coordination, motivation, and the response to environmental stress. However, the mechanisms involved in their subtype specification are not well understood. In this study, we examined the states of forebrain dopaminergic (DA) and serotonergic (5HT) neurons in larval and adult zebrafish of wild type and the too few mutant. The majority of DA and 5HT neuronal subgroups that were found in adults were established in the 6-day old larval zebrafish. Rather than affecting all monoaminergic neurons in the forebrain, selective subgroups of these neurons are reduced in the too few mutant, starting from the larval stage. Taken together, our study establishes that similar to DA neurons, distinct subtypes of 5HT neurons exist in larval as well as adult zebrafish. The development of a subset of these monoaminergic neurons is dependent on the too few gene product. Thus, this mutant is potentially important for understanding the development as well as the function of forebrain DA and 5HT neurons.

Section snippets

Experimental procedures

Heterozygous female zebrafish carrying the tof mutation were crossed with heterozygous males. Both sexes were genotyped by polymerase chain reaction (PCR) using a small piece of tissue from the tail. F2 progeny was raised at 28.5 °C in egg water (0.2 g Instant Ocean salts, 0.12 g CaSO4/l water). In order to investigate the defects in the DA system of the tof mutant we used 6-day old animals (seven homozygous, and nine wild type siblings) as well as adults (five homozygous and five wild type

Results

In the following text, the cells or groups that showed a positive immunostaining for TH, 5HT, or PCNA are called TH cells, 5HT cells, or PCNA cells, respectively.

5HT cell groups in the wild type zebrafish at 6d

In this study, we have reported for the first time the distribution of 5HT cells in the developing zebrafish forebrain. However, there are several publications describing the 5HT system in other teleosts (Ekström and van Veen, 1984, Ekström et al., 1995, Reiner and Northcutt, 1992, Ebbesson et al., 1992, Corio et al., 1991, Meek et al., 1989) or adult zebrafish (Kaslin and Panula, 2001). We used the latter study in order to compare the young and adult brain and find that all 5HT groups of the

Acknowledgements

We thank Drs. Gil Levkowitz and Arnon Rosenthal (Rinat Neuroscience Corporation, CA, USA) for communicating the method of genotyping tof mutant prior to publication, and Kayta Kobayashi (Biopharmaceutical Sciences, University of California, San Francisco, CA, USA) for assistance with genotyping. This work is financially supported by the Howard Hughes Medical Institute Biomedical Research Support Program grant to UCSF (5300246). S.G. is a Searle Scholar, Burroughs Wellcome Fund New Investigator,

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