An article published this year in “CELLULAR PHYSIOLOGY AND BIOCHEMISTRY”
using our Annexin V-FITC
Apoptosis Detection Kit, by our customers from Institute for Maternal and Child Health – IRCCS
“Burlo Garofolo” – Trieste, University of Trieste, Trieste, Italy, in the study
of Lack of Prenylated Proteins, Autophagy Impairment and Apoptosis in SH-SY5Y
Neuronal Cell Model of Mevalonate Kinase Deficiency. Congrats and Thanks.
Summary:
Background/Aims: Mevalonate Kinase Deficiency (MKD), is a hereditary disease due to
mutations in mevalonate kinase gene (MVK). MKD has heterogeneous
clinical phenotypes: the correlation between MVK mutations and MKD
clinical phenotype is still to be fully elucidated. Deficiency of prenylated
proteins has been hypothesized as possible MKD pathogenic mechanism. Based on
this hypothesis and considering that neurologic impairment characterizes
Mevalonic Aciduria (MA), the most severe form of MKD, we studied the effects of
I268T and N301T MVK mutations on protein prenylation, autophagy and
programmed cell death in SH-SY5Y neuroblastoma cell lines. Methods: SH-SY5Y
cells were transiently transfected, with the pCMV-6 plasmid containing MVK wild
type and the two mutated sequences. Protein prenylation levels were evaluated
using GFP-RhoA-F to assess farnesylation, and GFP-RhoA to evaluate
geranylgeranylation; autophagy was measured by evaluating LC3 and p62 protein
levels, while Annexin V-FITC and Propidium Iodide staining allowed apoptosis
detection. Results: MVK mutants’ over-expression causes
decreased levels of farnesylation and geranylgeranylation, and also increased
LC3 lipidation in SH-SY5Y, with concomitant p62 accumulation. Treatment with
bafilomycin A1 (an inhibitor of vacuolar H+-ATPase, a late autophagy inhibitor)
further increase LC3-II and p62 levels, suggesting that degradation of
autophagolysosome could be impaired. SH-SY5Y, with both MVK mutants,
showed apoptosis increase; the presence of N301T associated with augmented cell
death. Conclusions: We hypothesize that mevalonate pathway
impairment causes alteration of farnesylation and geranylgeranylation proteins
and alteration of the autophagic flux; these changes can induce apoptosis,
possibly more relevant in the presence of N301T mutation.
Reference:
Product link:
FITC Annexin V Apoptosis Detection Kit: http://www.immunostep.com/apoptosis-tools/3741-anxvkf-100t.html
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