Tar DNA-binding protein-43 (TDP-43) regulates axon growth in vitro and in vivo.

Intracellular inclusions of the TAR-DNA binding protein 43 (TDP-43) have been reported in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD-TDP). Rare mutations in TARDBP have been linked to both ALS and FTD-TDP suggesting that TDP-43 dysfunction is mechanistic in causing disease. TDP-43 is a predominantly nuclear protein with roles in regulating RNA transcription, splicing, stability and transport. In ALS, TDP-43 aberrantly accumulates in the cytoplasm of motor neurons where it forms aggregates. However it has until recently been unclear whether the toxic effects of TDP-43 involve recruitment to motor axons, and what effects this might have on axonal growth and integrity. Here we use chick embryonic motor neurons, in vivo and in vitro, to model the acute effects of TDP-43. We show that wild-type and two TDP-43 mutant proteins cause toxicity in chick embryonic motor neurons in vivo. Moreover, TDP-43 is increasingly mislocalised to axons over time in vivo, axon growth to peripheral targets is truncated, and expression of neurofilament-associated antigen is reduced relative to control motor neurons. In primary spinal motor neurons in vitro, a progressive translocation of TDP-43 to the cytoplasm occurs over time, similar to that observed in vivo. This coincides with the appearance of cytoplasmic aggregates, a reduction in the axonal length, and cellular toxicity, which was most striking for neurons expressing TDP-43 mutant forms. These observations suggest that the capacity of spinal motor neurons to produce and maintain an axon is compromised by dysregulation of TDP-43 and that the disruption of cytoskeletal integrity may play a role in the pathogenesis of ALS and FTD-TDP.

The surface ectoderm of the chick embryo exhibits dynamic variation in its response to neurogenic signals.

The epibranchial placodes are specialized areas of surface ectoderm that make a vital contribution to the peripheral nervous system, producing sensory neurons of the cranial ganglia. They have long been characterized as a series of patches of thickened ectoderm in the vicinity of each pharyngeal cleft. We have previously demonstrated that Sox3 is not only expressed in these structures but also marks a larger, earlier domain. Here we demonstrate that neurons are produced from the Sox3-positive ectoderm that lies outside of the classically-defined epibranchial placodes. Our data show that these regions contribute neurons to the cranial ganglia, but then cease producing neurons as they lose Sox3 expression. We further demonstrate that the ectoderm in these regions is responsive to extracellular or intracellular stimuli that initiate aspects of neuronal differentiation. This response to neurogenic stimuli is lacking in regions of ectoderm distant from the normal sites of neurogenesis and the response to constitutively active Bmp receptor in particular, disappears coincident with loss of Sox3 expression. Finally, we show that a dominant repressor form of Sox3 blocks the ability of the ectoderm to undergo neurogenesis. Thus, Sox3 appears to be essential for the neurogenic capacity of surface ectoderm exhibited by the epibranchial placodes.