Organotypic slice culture model demonstrates interneuronal spreading of alpha-synuclein aggregates

Elfarrash, Sara; Jensen, Nanna Møller; Ferreira, Nelson; Betzer, Cristine; Thevathasan, Jervis Vermal; Diekmann, Robin; Adel, Mohamed; Omar, Nor Asiah; Boraie, Mohamed Z.; Gad, Sabry; Ries, Jonas; Kirik, Deniz; Nabavi, Sadegh; Jensen, Poul Henning

doi:10.1101/681064

Cited by 3 publications

(14 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Organotypic hippocampal slices were prepared from 5-7-day-old C57Bl6 pups according to Stoppini et al 1991 [26] with slight modifications as described in [13]. Briefly, the hippocampi were extracted in carbonated low Na cerebrospinal fluid (CSF) (1 mM CaCl 2 , 10 mM D-glucose, 4 mM KCl, 5 mM MgCl 2 , 26 mM NaHCO 3 , 234 mM sucrose and 0.1% phenol red solution) and coronal slices of 400 µm were made using a tissue chopper (Stoelting, #51425).…”

Section: Preparation Of Organotypic Hippocampal Slice Cultures (Ohscs)mentioning

confidence: 99%

“…PFFs were produced from monomeric human recombinant α-syn with residue serine-129 shifted to alanine (S129A). S129A PFF production, characterization and validation of efficient aggregation was done as described in [13]. The usage of S129A PFFs that are nonphosphorylatable at S129 ensures that only the endogenously expressed α-syn is detected when antibodies against pS129 are used during analysis.…”

Section: Microinjection Of Ohscs With S129a Pffsmentioning

confidence: 99%

“…The pipette was loaded using Eppendorf microloader pipette tips (ThermoFisher). A final volume of 0.1 µL of either S129A PFFs (1 mg/mL) or PBS was injected at DG under microscopic guidance as described in [13].…”

Section: Microinjection Of Ohscs With S129a Pffsmentioning

confidence: 99%

“…For each group, ten slices were analyzed. Protein quantification, sequential fractionation into RIPA-soluble andinsoluble fractions and subsequent immunoblotting was carried out as described in [13].…”

Section: Immunoblotting and Sequential Biochemical Fractionation Of Ohscsmentioning

confidence: 99%

“…Despite the advantages of pS129 as a biomarker for pathological α-syn aggregates, its pathophysiological role has been contested [6], [7], [8], [9], [10]. Some studies demonstrate an increased toxicity associated to S129-phosphorylation [8], [9], [11], whereas others suggests it has no role in α-syn aggregation or toxicity [12], [13] or even being a protective modification [6], [7].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Polo-like kinase 2 inhibition reduces serine-129 phosphorylation of physiological nuclear alpha-synuclein but not of the aggregated alpha-synuclein

Elfarrash

Jensen

Ferreira

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Accumulation of aggregated alpha-synuclein (α-syn) is believed to play a pivotal role in the pathophysiology of Parkinson’s disease (PD) and other synucleinopathies. α-Syn is a key constituent protein of Lewy pathology, and α-syn phosphorylated at serine-129 (pS129) constitutes more than 90% of α-syn in Lewy bodies and hence, it is used extensively as a pathological marker for the aggregated form of α-syn. However, the exact role of pS129 remains controversial as well as the kinase(s) responsible for the phosphorylation. In this study, we investigated the effect of Polo-like kinase 2 (PLK2) inhibition on formation of pS129 using ex-vivo organotypic brain slice model of synucleinopathy. Our data demonstrated that PLK2 inhibition has no effect on α-syn aggregation, pS129 or inter-neuronal spreading of the aggregated α-syn seen in the organotypic slices. Instead, PLK2 inhibition reduced the soluble nuclear pS129 level confined in the nuclei. The same finding was replicated in an in-vivo mouse models of templated α-syn aggregation and human dopaminergic neurons, suggesting that PLK2 is more likely to be involved in S129 phosphorylation of soluble non-pathology related fraction of α-syn. We also demonstrated that reduction of nuclear pS129 but not the aggregates specific pS129 following PLK2 inhibition for a short time before sample collection improves the signal to noise ratio when quantifying pS129 aggregate pathology.

show abstract

Section: Preparation Of Organotypic Hippocampal Slice Cultures (Ohscs)mentioning

confidence: 99%

Section: Microinjection Of Ohscs With S129a Pffsmentioning

confidence: 99%

Section: Microinjection Of Ohscs With S129a Pffsmentioning

confidence: 99%

Section: Immunoblotting and Sequential Biochemical Fractionation Of Ohscsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Polo-like kinase 2 inhibition reduces serine-129 phosphorylation of physiological nuclear alpha-synuclein but not of the aggregated alpha-synuclein

Elfarrash

Jensen

Ferreira

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Brain‐Penetration and Neuron‐Targeting DNA Nanoflowers Co‐Delivering miR‐124 and Rutin for Synergistic Therapy of Alzheimer's Disease

Ouyang

Zhu

et al. 2022

Small

View full text Add to dashboard Cite

tau hyperphosphorylation, mitochondrial dysfunction and oxidative stress, and neuroinflammatory response. [3] Among them, the amyloid theory that focuses on excessive deposition of β-amyloid (Aβ) has been extensively studied with wide acceptance. [4] In this theory, imbalance of Aβ metabolism and catabolism in the central nervous system (CNS) has been regarded as the main reason for AD-like pathology. [5] Aβ is the product of a proteolytic process of a transmembrane protein, amyloid precursor protein (APP), by β-secretase 1 (BACE1, β-site APP cleavage enzyme 1) and the γ-secretase. [6] The overgeneration and abnormal aggregation of Aβ 1-42 can cause the collapse of dendritic spines and neuronal loss, induce oxidative stress, and promote inflammatory by activating microglia, which in turn exacerbates the pathogenesis of AD. [7] Several strategies focus on blocking Aβ aggregation and alleviating Aβ-mediated neurotoxicity have been developed, including aggregation inhibition, copper chelation, Aβ photo-oxidation, fibril thermal dissociation, and Aβ degradation. [8] However, many Aβ targeting pharmacological treatments failed to slow down genitive decline or improve global functioning in phase 3 clinical trials although with definite efficacy at animal model. [9] Besides the pathological discrepancy between real clinical patient and AD animal model, [10] another important reason is that AD is a multifactorial disease and therapy targeting Aβ alone is far more enough. [11] Despite significant Alzheimer disease (AD) is the leading cause of dementia that affects millions of old people. Despite significant advances in the understanding of AD pathobiology, no disease modifying treatment is available. MicroRNA-124 (miR-124) is the most abundant miRNA in the normal brain with great potency to ameliorate AD-like pathology, while it is deficient in AD brain. Herein, the authors develop a DNA nanoflowers (DFs)-based delivery system to realize exogenous supplementation of miR-124 for AD therapy. The DFs with well-controlled size and morphology are prepared, and a miR-124 chimera is attached via hybridization. The DFs are further modified with RVG29 peptide to simultaneously realize brain-blood barrier (BBB) penetration and neuron targeting. Meanwhile, Rutin, a small molecular ancillary drug, is co-loaded into the DFs structure via its intercalation into the double stranded DNA region. Interestingly, Rutin could synergize miR-124 to suppress the expression of both BACE1 and APP, thus achieving a robust inhibition of amyloid β generation. The nanosystem could pro-long miR-124 circulation in vivo, promote its BBB penetration and neuron targeting, resulting in a significant increase of miR-124 in the hippocampus of APP/PS1 mice and robust therapeutic efficacy in vivo. Such a bio-derived therapeutic system shows promise as a biocompatible nanomedicine for AD therapy.

show abstract

The history of Danish neuroscience

Paulson

Schousboe

Hultborn

2023

Eur J of Neuroscience

View full text Add to dashboard Cite

The history of Danish neuroscience starts with an account of impressive contributions made at the 17th century. Thomas Bartholin was the first Danish neuroscientist, and his disciple Nicolaus Steno became internationally one of the most prominent neuroscientists in this period. From the start, Danish neuroscience was linked to clinical disciplines. This continued in the 19th and first half of the 20th centuries with new initiatives linking basic neuroscience to clinical neurology and psychiatry in the same scientific environment. Subsequently, from the middle of the 20th century, basic neuroscience was developing rapidly within the preclinical university sector. Clinical neuroscience continued and was even reinforced during this period with important translational research and a close co‐operation between basic and clinical neuroscience. To distinguish ‘history’ from ‘present time’ is not easy, as many historical events continue in present time. Therefore, we decided to consider ‘History’ as new major scientific developments in Denmark, which were launched before the end of the 20th century. With this aim, scientists mentioned will have been born, with a few exceptions, no later than the early 1960s. However, we often refer to more recent publications in documenting the developments of initiatives launched before the end of the last century. In addition, several scientists have moved to Denmark after the beginning of the present century, and they certainly are contributing to the present status of Danish neuroscience—but, again, this is not the History of Danish neuroscience.

show abstract

Organotypic slice culture model demonstrates interneuronal spreading of alpha-synuclein aggregates

Cited by 3 publications

References 42 publications

Polo-like kinase 2 inhibition reduces serine-129 phosphorylation of physiological nuclear alpha-synuclein but not of the aggregated alpha-synuclein

Polo-like kinase 2 inhibition reduces serine-129 phosphorylation of physiological nuclear alpha-synuclein but not of the aggregated alpha-synuclein

Brain‐Penetration and Neuron‐Targeting DNA Nanoflowers Co‐Delivering miR‐124 and Rutin for Synergistic Therapy of Alzheimer's Disease

The history of Danish neuroscience

Contact Info

Product

Resources

About