Characterization of GFP-tagged GnRH-containing terminalis neurons in transgenic zebrafish

The terminalis nerve (TN) has been described in all vertebrate species, in which it plays important roles in animal behavior and physiology. In teleost fish, the TN is located in the olfactory bulb and in its nerve tract. Here, we report a study on the characterization of the TN cell development, axon projection and physiology in zebrafish (Danio rerio). We have generated several lines of transgenic zebrafish [Tg (GnRH-3::GFP)] that express GFP in the TN cells. The transgenes are expressed under the transcriptional control of the zebrafish GnRH-3 promoter. During development, the first GFP-positive TN cell was identified at approximately 34 h post-fertilization (hpf). By 38 hpf, several clusters of TN cells were identified in the olfactory bulb and olfactory nerve tract. In the olfactory bulb, the TN cells projected axons caudally. In the forebrain, some of the TN axons extended caudally, but most crossed the midline of the brain at the commissural anterior. In the midbrain, some of the TN axons extended dorsally towards the tectum, whereas other axons extended caudally, or extended ventrally to the optic nerve where they entered the neural retina. We also examined the cell membrane property of the TN cells. Using patch-clamp techniques, we recorded spontaneous and evoked action potentials from isolated TN cells. We examined the expression of glutamate receptors in the TN cells. The data shed light on the mechanisms of TN function in the nervous system and in the regulation of animal physiology.