Lack of Neuroprotective Effect of Celastrol Under Conditions of Proteasome Inhibition by Lactacystin in In Vitro and In Vivo Studies: Implications for Parkinson’s Disease

Konieczny, Jolanta; Jantas, Danuta; Lenda, Tomasz; Domin, Helena; Czarnecka, Anna; Kuter, Katarzyna; Śmiałowska, Maria; Lasoń, Władysław; Lorenc‐Koci, Elżbieta

doi:10.1007/s12640-014-9477-9

Cited by 35 publications

(23 citation statements)

References 80 publications

(111 reference statements)

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“…In this study, we have characterized a mouse model of PD that is based on direct proteasomal inhibition in the SN of young and adult mice. Although LC has been used as a PD model in earlier studies either by injecting it into the medial forebrain bundle or striatum (Fornai et al 2003;Vernon et al 2011;Shen et al 2013;Konieczny et al 2014a), and recently in mouse (Bentea et al 2015;Kumar et al 2015) and rat (Konieczny et al 2014b;Harrison et al 2015;Pienaar et al 2015) SN, to our knowledge this is the first time when LC injection has also been used in adult mice and wide-spread neuroinflammation has been observed and thoroughly analyzed. The model shows several well-established features of PD -motor deficits, nigrostriatal dopamine loss and aSyn accumulation as the most important, and we indeed detected differences in response between young and adult mice.…”

Section: Discussionmentioning

confidence: 99%

Nigral injection of a proteasomal inhibitor, lactacystin, induces widespread glial cell activation and shows various phenotypes of Parkinson’s disease in young and adult mouse

et al. 2017

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Proteinaceous inclusions, called Lewy bodies, are used as a pathological hallmark for Parkinson's disease (PD). Lewy bodies contain insoluble α-synuclein (aSyn) and many other ubiquitinated proteins, suggesting a role for protein degradation system failure in the PD pathogenesis. Indeed, proteasomal dysfunction has been linked to PD but commonly used in vivo toxin models, such as 6-OHDA or MPTP, do not have a significant effect on the proteasomal system or protein aggregation. Therefore, we wanted to study the characteristics of a proteasomal inhibitor, lactacystin, as a PD model on young and adult mice. To study this, we performed stereotactic microinjection of lactacystin above the substantia nigra pars compacta in young (2 month old) and adult (12-14 month old) C57Bl/6 mice. Motor behavior was measured by locomotor activity and cylinder tests, and the markers of neuroinflammation, aSyn, and dopaminergic system were assessed by immunohistochemistry and HPLC. We found that lactacystin induced a Parkinson's disease-like motor phenotype 5-7 days after injection in young and adult mice, and this was associated with widespread neuroinflammation based on glial cell markers, aSyn accumulation in substantia nigra, striatal dopamine decrease, and loss of dopaminergic cell bodies in the substantia nigra and terminals in the striatum. When comparing young and adult mice, adult mice were more sensitive for dopaminergic degeneration after lactacystin injection that further supports the use of adult mice instead of young when modeling neurodegeneration. Our data showed that lactacystin is useful in modeling various aspects of Parkinson's disease, and taken together, our findings emphasize the role of a protein degradation deficit in Parkinson's disease pathology, and support the use of proteasomal inhibitors as Parkinson's disease models.

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Section: Discussionmentioning

confidence: 99%

Nigral injection of a proteasomal inhibitor, lactacystin, induces widespread glial cell activation and shows various phenotypes of Parkinson’s disease in young and adult mouse

et al. 2017

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“…Celastrol has low toxicity and is in fact being explored as a therapeutic due to its positive effects on the immune system (95)(96)(97) . The concentration of celastrol that is sufficient to toggle Gal4-MYB activity in polycomb chromatin is well below the reported LD50 values for this drug (98)(99)(100)(101)(102) .…”

Section: Discussionmentioning

confidence: 62%

Components from the human c-myb transcriptional regulation system reactivate epigenetically repressed transgenes

Barrett

McCracken

Elmer

et al. 2018

Preprint

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Epigenetic silencing of transgenes has been a persistent challenge for mammalian cell engineering. Foreign DNA can be incorporated into closed chromatin before and after it has been integrated into a host cell's genome. To identify elements that mitigate epigenetic silencing, we tested components from the c-myb and NF-kB transcriptional regulation systems in transiently transfected DNA and at chromosomally integrated transgenes in PC-3 and HEK293 cells. DNA binding sites for MYB (c-myb) placed upstream of a minimal promoter strongly enhanced expression from transiently transfected plasmid DNA. We targeted p65 and MYB fusion proteins to chromosomal transgenes that were silenced by ectopic Polycomb chromatin or by uncharacterized endogenous chromatin. Transient expression of Gal4-MYB induced sustained activation of the Polycomb-silenced UAS-Tk-luciferase transgene. We used custom guide RNAs and dCas9-MYB to target MYB to different sites. Transgene activation within ectopic Polycomb chromatin required proximity of dCas9-MYB to the transcriptional start site, while activation at the naturally repressed transgene was position-independent. Our report is the first to demonstrate the use of MYB in the context of the CRISPR-activation system. The results demonstrate that DNA elements and fusion proteins derived from c-myb can mitigate epigenetic silencing to improve transgene expression in engineered cell lines.

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“…Celastrol has low toxicity and is in fact being explored as a therapeutic due to its positive effects on the immune system [92][93][94]. The concentration of celastrol that is sufficient to toggle Gal4-MYB activity in polycomb chromatin is well below the reported LD50 values for this drug [95][96][97][98][99].…”

Section: Discussionmentioning

confidence: 99%

Components from the Human c-myb Transcriptional Regulation System Reactivate Epigenetically Repressed Transgenes

Barrett

McCracken

Elmer

et al. 2020

IJMS

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A persistent challenge for mammalian cell engineering is the undesirable epigenetic silencing of transgenes. Foreign DNA can be incorporated into closed chromatin before and after it has been integrated into a host cell’s genome. To identify elements that mitigate epigenetic silencing, we tested components from the c-myb and NF-kB transcriptional regulation systems in transiently transfected DNA and at chromosomally integrated transgenes in PC-3 and HEK 293 cells. DNA binding sites for MYB (c-myb) placed upstream of a minimal promoter enhanced expression from transiently transfected plasmid DNA. We targeted p65 and MYB fusion proteins to a chromosomal transgene, UAS-Tk-luciferase, that was silenced by ectopic Polycomb chromatin complexes. Transient expression of Gal4-MYB induced an activated state that resisted complete re-silencing. We used custom guide RNAs and dCas9-MYB to target MYB to different positions relative to the promoter and observed that transgene activation within ectopic Polycomb chromatin required proximity of dCas9-MYB to the transcriptional start site. Our report demonstrates the use of MYB in the context of the CRISPR-activation system, showing that DNA elements and fusion proteins derived from c-myb can mitigate epigenetic silencing to improve transgene expression in engineered cell lines.

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Lack of Neuroprotective Effect of Celastrol Under Conditions of Proteasome Inhibition by Lactacystin in In Vitro and In Vivo Studies: Implications for Parkinson’s Disease

Cited by 35 publications

References 80 publications

Nigral injection of a proteasomal inhibitor, lactacystin, induces widespread glial cell activation and shows various phenotypes of Parkinson’s disease in young and adult mouse

Nigral injection of a proteasomal inhibitor, lactacystin, induces widespread glial cell activation and shows various phenotypes of Parkinson’s disease in young and adult mouse

Components from the human c-myb transcriptional regulation system reactivate epigenetically repressed transgenes

Components from the Human c-myb Transcriptional Regulation System Reactivate Epigenetically Repressed Transgenes

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