A therapeutic target for Parkinson's disease and its application
A Parkinson's disease, short peptide technology, applied to Parkinson's disease treatment targets and their application fields, can solve problems such as increasing gait and cognitive risk
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Embodiment 1
[0034] Example 1 Demonstration experiment of the existence of monosynaptic connection of STN-ANT
[0035] The inventors found that the nuclei downstream of the STN also include the unreported anterior thalamic nucleus (ANT) by virus tracing method.
[0036] First, in terms of anatomical connections, after STN injection with a fluorescently tagged anterograde non-transsynaptic virus (rAAV2 / 9-CaMKIIα-EYFP-WPRE-pA, BrainVTA PT-0102), brain slices were excised to observe ANT nuclei fibers labeled with the fluorescent label EYFP, such as figure 1 Shown in A; further, by means of the Cre-LoxP recombinase system, the cis-transmonosynaptic virus (rAAV2 / 1-CaMKIIα-Cre-WPRE-pA, BrainVTA PT-0220) with the Cre enzyme gene was injected into the STN At the same time, the virus with LoxP system (rAAV-Ef1α-Dio-EYFP-WPRE-pA, BrainVTA PT-0012) was injected into ANT, when Cre enzyme entered ANT from STN anterogradely across monosynapses, the Lox site in ANT could be transferred to ANT. At the p...
Embodiment 2
[0037] Example 2 Increased plasticity of the STN-ANT loop on the injured side in unilateral Parkinson's disease model mice
[0038]Construction of unilateral Parkinson's disease mouse model: unilateral striatal 6-OHDA injection (Ref. Francardo V, Recchia A, Popovic N, et al. 2011. Impact of the lesion procedure on the profiles of motor impairment and molecular responsiveness to L-DOPA in the 6-hydroxydopamine mouse model of Parkinson's disease. Neurobiol. Dis. 42, 327-340), to establish a unilateral Parkinson's disease mouse model, which was successfully modeled by apomorphine-induced spinning behavior after 7 to 10 days .
[0039] The successfully modeled mice were injected into STN with a virus (rAAV-CaMKIIα-hChR2(H134R)-mCherry-WPRE-pA, Brain VTA PT-0279) with the light-sensing protein ChR2, and electrophysiologically coupled by optogenetics. Methods The α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor (α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid) w...
Embodiment 3
[0049] Example 3 TAT-AMPA-S845 peptide modulates changes in synaptic plasticity
[0050] According to previous literature reports, phosphorylation of AMPA receptors in the upper membrane is a major way to affect synaptic strength, and the increase in the upper membrane caused by phosphorylation of the AMPAR isoform GluR1 at the S845 site has received extensive attention. In order to explore this phenomenon, Western blot (Western Blot, WB) was used to measure AMPAR and its phosphorylation sites in the ipsilateral (Ips) and non-injured (contralateral, Con) sides of PD in ANT nuclei. (S831, S845) changes, the inventor found that only the phosphorylation of S845 site increased, and other phosphorylation sites did not see changes. This result indicates that the increase in phosphorylation of the AMPAR isoform GluR1 S845 of the ANT nucleus in the PD state leads to an increase in the membrane of AMPAR, which leads to changes in plasticity. In vivo, phosphorylation at S845 is mediate...
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