Please go through this article and make a very detailed list

Please go through this article and make a very detailed list of all techniques used by the researchers and what the technique was used to discover or investigate. Be as specific as possible. Copy the link below for the article.

article: https://www.dropbox.com/s/29hb87u75kayi9z/Abeliovich-Gitler_Defects%20in%20trafficking-2017.pdf?dl=0

Solution

The techniques use by researchers are given as:

1) Dopamine replacement therapies, such as the dopamine precursor levodopa, typically lead to the relief of symptoms but lose their potency as the disease progresses. It was discovered because:

Clinical manifestations of PD(Parkinsons disease) include motor deficits such as rigidity, slowness in movement (bradykinesia), postural instability and a characteristic tremor at rest. The motor symptoms of PD result from the selective loss of dopaminergic neurons in the pars compacta of the substantia nigra (SN) in the midbrain, as well as their axon terminals, which project to the dorsal striatum.

2) Ubiquitin monomers or polyubiquitin chains are required for the proper intracellular trafficking and disposal of many proteins in cells, and the accumulation of ubiquitin is therefore consistent with defects in trafficking and disposal pathways. This technique was discovered because:

A neuropathological hallmark of PD is the presence of intraneuronal proteinaceous inclusions, termed Lewy bodies (LBs) or Lewy neurites. These structures are enriched in filamentous forms of the synaptic protein -synuclein as well as the small regulatory protein ubiquitin.

3) Defective trafficking to the lysosome is the most prominent mechanism of pathogenesis that links PD-associated genetic variants and mutations, and it therefore has high priority as a therapeutic target.

4)The emerging role of VPS35 mutations and retromer-complex function in PD means that approaches that boost retromer activity could be a promising therapeutic avenue. The technique was discoered because:

Strategies to selectively improve the function or delivery of glucosylceramidase, including functional activators of the enzyme or molecular chaperones that stabilize its structure have been considered, but their clinical efficacy has not been described. Because several of the most common pathogenic mutations in LRRK2, including G2019S, result in promiscuous LRRK2 kinase activity, approaches to inhibit such activity are being pursued.However, the relevance of altered LRRK2 kinase activity to the development of PD is unresolvedand, paradoxically, inhibitors of LRRK2 kinase activity have been associated with lysosomal disorders in vivo.

5) Improved autophagic flux, for example, through inhibition of the serine–threonine-protein kinase mTOR with rapamycin, has been evaluated, but such a strategy could be counterproductive if the main defect is downstream in the pathway at the fusion of autophagosomes to lysosomes.

6) A therapeutic small molecule called NAB2, which might broadly suppress defects in endosomal trafficking that occur downstream of an excess of -synuclein, has been described using a yeast-based screen.

7) Inhibitors of oxidative or nitrosative damage, which might promote toxic structures, have also been proposed

8) Therapeutic efforts that target -synuclein might be broadly effective if the protein is an essential downstream effector of PD-associated neuron loss, regardless of the initial insult. Alternatively, it is possible that mechanisms that are independent of -synuclein, and perhaps related to trafficking dysfunction, drive PD-associated neurotoxicity in most cases.