A zebrafish model exemplifies the long preclinical period of motor neuron disease

A zebrafish model exemplifies the long preclinical period of motor neuron diseas

Graphene oxide nanosheets modulate spinal glutamatergic transmission and modify locomotor behaviour in an in vivo zebrafish model

Graphene oxide (GO), an oxidised form of graphene, is widely used for biomedical applications, due to its dispersibility in water and simple surface chemistry tunability. In particular, small (less than 500 nm in lateral dimension) and thin (1-3 carbon monolayers) graphene oxide nanosheets (s-GO) have been shown to selectively inhibit glutamatergic transmission in neuronal cultures in vitro and in brain explants obtained from animals injected with the nanomaterial. This raises the exciting prospect that s-GO can be developed as a platform for novel nervous system therapeutics. It has not yet been investigated whether the interference of the nanomaterial with neurotransmission may have a downstream outcome in modulation of behaviour depending specifically on the activation of those synapses. To address this problem we use early stage zebrafish as an in vivo model to study the impact of s-GO on nervous system function. Microinjection of s-GO into the embryonic zebrafish spinal cord selectively reduces the excitatory synaptic transmission of the spinal network, monitored in vivo through patch clamp recordings, without affecting spinal cell survival. This effect is accompanied by a perturbation in the swimming activity of larvae, which is the locomotor behaviour generated by the neuronal network of the spinal cord. Such results indicate that the impact of s-GO on glutamate based neuronal transmission is preserved in vivo and can induce changes in animal behaviour. These findings pave the way for use of s-GO as a modulator of nervous system function

GCH1 deficiency activates brain innate immune response and impairs tyrosine hydroxylase homeostasis

The Parkinson’s disease (PD) risk gene GTP cyclohydrolase 1 (GCH1) catalyzes the rate-limiting step in tetrahydrobiopterin (BH4) synthesis, an essential cofactor in the synthesis of monoaminergic neurotransmitters. To investigate the mechanisms by which GCH1 deficiency may contribute to PD, we generated a loss of function zebrafish gch1 mutant (gch1-/-), using CRISPR/Cas technology. gch1-/- zebrafish develop marked monoaminergic neurotransmitter deficiencies by 5 dpf, movement deficits by 8 dpf and lethality by 12 dpf. Tyrosine hydroxylase protein levels were markedly reduced without loss of ascending dopaminergic (DAergic) neurons. L-Dopa treatment of gch1-/- larvae improved survival without ameliorating the motor phenotype. RNAseq of gch1-/- larval brain tissue identified highly upregulated transcripts involved in innate immune response. Subsequent experiments provided morphological and functional evidence of microglial activation in gch1-/-. The results of our study suggest that GCH1 deficiency may unmask early, subclinical parkinsonism and only indirectly contribute to neuronal cell death via immune-mediated mechanisms. Our work highlights the importance of functional validation for GWAS risk factors and further emphasises the important role of inflammation in the pathogenesis of PD

GCH1 deficiency activates brain innate immune response and impairs tyrosine hydroxylase homeostasis

The Parkinson's disease (PD) risk gene GTP cyclohydrolase 1 (GCH1) catalyzes the rate-limiting step in tetrahydrobiopterin (BH4) synthesis, an essential cofactor in the synthesis of monoaminergic neurotransmitters. To investigate the mechanisms by which GCH1 deficiency may contribute to PD, we generated a loss of function zebrafish gch1 mutant (gch1–/–), using CRISPR/Cas technology. gch1–/– zebrafish develop marked monoaminergic neurotransmitter deficiencies by 5 d postfertilization (dpf), movement deficits by 8 dpf and lethality by 12 dpf. Tyrosine hydroxylase (Th) protein levels were markedly reduced without loss of ascending dopaminergic (DAergic) neurons. L-DOPA treatment of gch1–/– larvae improved survival without ameliorating the motor phenotype. RNAseq of gch1–/– larval brain tissue identified highly upregulated transcripts involved in innate immune response. Subsequent experiments provided morphologic and functional evidence of microglial activation in gch1–/–. The results of our study suggest that GCH1 deficiency may unmask early, subclinical parkinsonism and only indirectly contribute to neuronal cell death via immune-mediated mechanisms. Our work highlights the importance of functional validation for genome-wide association studies (GWAS) risk factors and further emphasizes the important role of inflammation in the pathogenesis of PD

Aminergic modulation of glycine release in a spinal network controlling swimming in Xenopus laevis

1. Neuromodulators can effect changes in neural network function by strengthening or weakening synapses between neurons via presynaptic control of transmitter release. We have examined the effects of two biogenic amines on inhibitory connections of a spinal rhythm generator in Xenopus tadpoles.2. Glycinergic inhibitory potentials occurring mid-cycle in motoneurons during swimming activity are reduced by 5-hydroxytryptamine (5-HT; serotonin) and enhanced by noradrenaline (NA). These opposing effects on inhibitory synaptic strength are mediated presynaptically where 5-HT decreases and Nrl increases the probability of glycine release from inhibitory terminals.3. The amines also have contrasting effects on swimming: 5-HT increased motor burst durations while NA reduced swimming frequency Aminergic modulation of glycinergic transmission mag thus control fundamental parameters of swimming and force the spinal network to generate opposite extremes of its spectrum of possible outputs.</p

Aminergic modulation of glycine release in a spinal network controlling swimming in Xenopus laevis

1. Neuromodulators can effect changes in neural network function by strengthening or weakening synapses between neurons via presynaptic control of transmitter release. We have examined the effects of two biogenic amines on inhibitory connections of a spinal rhythm generator in Xenopus tadpoles.2. Glycinergic inhibitory potentials occurring mid-cycle in motoneurons during swimming activity are reduced by 5-hydroxytryptamine (5-HT; serotonin) and enhanced by noradrenaline (NA). These opposing effects on inhibitory synaptic strength are mediated presynaptically where 5-HT decreases and Nrl increases the probability of glycine release from inhibitory terminals.3. The amines also have contrasting effects on swimming: 5-HT increased motor burst durations while NA reduced swimming frequency Aminergic modulation of glycinergic transmission mag thus control fundamental parameters of swimming and force the spinal network to generate opposite extremes of its spectrum of possible outputs.</p