The arrangement of the early nerve connections in the embryonic vertebrate brain follows a well-conserved pattern, forming the early axon scaffold. The early axon tracts have been described in a number of anamniote species and in mouse, but a detailed analysis in chick is lacking. We have used immunostaining, axon tracing and in situ hybridisation to analyse the development of the early axon scaffold in the embryonic chick brain in relation to the neuromeric organisation of the brain. The first tract to be formed is the medial longitudinal fascicle (MLF), shortly followed by the tract of the postoptic commissure to pioneer the ventral longitudinal tract system. The MLF was found to originate from three different populations of neurones located in the diencephalon. Neurones close to the dorsal midline of the mesencephalon establish the descending tract of the mesencephalic nucleus of the trigeminus. Their axons pioneer the lateral longitudinal tract. At later stages, the tract of the posterior commissure emerges in the caudal pretectum as the first transversal tract. It is formed by dorsally projecting axons from neurones located in the ventral pretectum, and by ventrally projecting axons from neurones located in the dorsal pretectum. The organisation of neurones and axons in the chick brain is similar to that described in the mouse, though tracts form in a different order and appear more clearly distinguished than in the mammalian model. Key words: early axon scaffold; medial longitudinal fascicle; prosomeric model; tract of the posterior commisure; tract of the postoptic commissure.

BackgroundThe generation of diverse neuronal types and subtypes from multipotent progenitors during development is crucial for assembling functional neural circuits in the adult central nervous system. It is well known that the Notch signalling pathway through the inhibition of proneural genes is a key regulator of neurogenesis in the vertebrate central nervous system. However, the role of Notch during hypothalamus formation along with its downstream effectors remains poorly defined.ResultsHere, we have transiently blocked Notch activity in chick embryos and used global gene expression analysis to provide evidence that Notch signalling modulates the generation of neurons in the early developing hypothalamus by lateral inhibition. Most importantly, we have taken advantage of this model to identify novel targets of Notch signalling, such as Tagln3 and Chga, which were expressed in hypothalamic neuronal nuclei.ConclusionsThese data give essential advances into the early generation of neurons in the hypothalamus. We demonstrate that inhibition of Notch signalling during early development of the hypothalamus enhances expression of several new markers. These genes must be considered as important new targets of the Notch/proneural network.

The establishment of a functional nervous system requires a highly orchestrated process of neural proliferation and differentiation. The evolutionary conserved Notch signaling pathway is a key regulator of this process, regulating basic helix-loop-helix (bHLH) transcriptional repressors and proneural genes. However, little is known about downstream Notch targets and subsequently genes required for neuronal specification. In this report, the expression pattern of Transgelin 3 (Tagln3), Chromogranin A (Chga) and Contactin 2 (Cntn2) was described in detail during early chick embryogenesis. Expression of these genes was largely restricted to the nervous system including the early axon scaffold populations, cranial ganglia and spinal motor neurons. Their temporal and spatial expression were compared with the neuronal markers Nescient Helix-Loop-Helix 1 (Nhlh1), Stathmin 2 (Stmn2) and HuC/D. We show that Tagln3 is an early marker for post-mitotic neurons whereas Chga and Cntn2 are expressed in mature neurons. We demonstrate that inhibition of Notch signaling during spinal cord neurogenesis enhances expression of these markers. This data demonstrates that Tagln3, Chga and Cntn2 represent strong new candidates to contribute to the sequential progression of vertebrate neurogenesis.