PINK1 and Alzheimer's and Multiple Sclerosis

I would like to highlight a recent publication referencing use of our PTEN-induced kinase/PINK1 Antibody.

Jack Van Horssen et al. investigated PINK1 expression in well-characterized brain samples derived from MS and AD individuals using immunohistochemistry.

Micha M.M. Wilhelmus, Susanne M.A. van der Pol, Quentin Jansen, Maarten E. Witte, Paul van der Valk, Annemieke J.M. Rozemuller, Benjamin Drukarch, Helga E. de Vries and Jack Van Horssen. Association of Parkinson disease-related protein PINK1 with Alzheimer disease and multiple sclerosis brain lesions. Free Radical Biology and Medicine. Volume 50, Issue 3, 1 February 2011, Pages 469-476.

Abstract: Mitochondrial dysfunction and oxidative stress are hallmarks of various neurological disorders, including multiple sclerosis (MS), Alzheimer disease (AD), and Parkinson disease (PD). Mutations in PINK1, a mitochondrial kinase, have been linked to the occurrence of early onset parkinsonism. Currently, various studies support the notion of a neuroprotective role for PINK1, as it protects cells from stress-mediated mitochondrial dysfunction, oxidative stress, and apoptosis. Because information about the distribution pattern of PINK1 in neurological diseases other than PD is scarce, we here investigated PINK1 expression in well-characterized brain samples derived from MS and AD individuals using immunohistochemistry. In control gray matter PINK1 immunoreactivity was observed in neurons, particularly neurons in layers IV–VI. Astrocytes were the most prominent cell type decorated by anti-PINK1 antibody in the white matter. In addition, PINK1 staining was observed in the cerebrovasculature. In AD, PINK1 was found to colocalize with classic senile plaques and vascular amyloid depositions, as well as reactive astrocytes associated with the characteristic AD lesions. Interestingly, PINK1 was absent from neurofibrillary tangles. In active demyelinating MS lesions we observed a marked astrocytic PINK1 immunostaining, whereas astrocytes in chronic lesions were weakly stained. Taken together, we observed PINK1 immunostaining in both AD and MS lesions, predominantly in reactive astrocytes associated with these lesions, suggesting that the increase in astrocytic PINK1 protein might be an intrinsic protective mechanism to limit cellular injury.

Immunohistochemistry

Immunohistochemistry was used to detect PINK1 immunostaining in temporal neocortex and white matter in AD, MS, and control subjects. Cryosections (5 μm) were air-dried and fixed in acetone for 10 min. Next, sections were incubated with an affinity-purified rabbit anti-PINK1 antibody (1:100; Neuromics, Edina, MN, USA) for 60 min at room temperature. Then, the slides were incubated with EnVision kit horseradish peroxidase-labeled anti-mouse/rabbit (DAKO, Glostrup, Denmark) for 30 min at room temperature and finally diaminobenzidine tetrachloride. Between incubation steps, sections were thoroughly washed with phosphate-buffered saline (PBS). After a short rinse in tap water sections were incubated with hematoxylin for 1 min and extensively washed with tap water for 10 min. Finally, sections were dehydrated with ethanol followed by xylol and mounted with Entellan (Merck, Darmstadt, Germany). All antibodies were diluted in PBS containing 0.1% bovine serum albumin (Boehringer–Mannheim, Germany), which also served as a negative control. Negative controls were essentially blank.

Image: In active lesions PINK1 immunostaining was intense in reactive astrocytes (arrows). Double labeling of PINK1 (green) with the astrocytic marker GFAP (red) demonstrated PINK1 expression in astrocytes (inset).

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Glutaredoxin 2 prevents aggregation of mutant SOD1

Our PTEN-induced kinase, PINK1 or PARK6 Antibody is an excellent marker for Amyotrophic Lateral Sclerosis (ALS) and Parkinson's Disease (PD) researchers.

Here's  new publication referencing use of this antibody:  Alberto Ferri, Paolo Fiorenzo, Monica Nencini, Mauro Cozzolino, Maria Grazia Pesaresi, Cristiana Valle, Sara Sepe, Sandra Moreno, and Maria Teresa Carrì. Glutaredoxin 2 prevents aggregation of mutant SOD1 in mitochondria and abolishes its toxicity.
Hum. Mol. Genet., first published on Sep 20, 2010 as doi: doi:10.1093/hmg/ddq383

Abstract:
Vulnerability of motoneurons in amyotrophic lateral sclerosis (ALS) arises from a combination of several mechanisms, including protein misfolding and aggregation, mitochondrial dysfunction and oxidative damage. Protein aggregates are found in motoneurons in models for ALS linked to a mutation in the gene coding for Cu,Zn superoxide dismutase (SOD1) and in ALS patients as well. Aggregation of mutant SOD1 in the cytoplasm and/or into mitochondria has been repeatedly proposed as a main culprit for the degeneration of motoneurons. It is, however, still debated whether SOD1 aggregates represent a cause, a correlate or a consequence of processes leading to cell death. We have exploited the ability of glutaredoxins (Grxs) to reduce mixed disulfides to protein thiols either in the cytoplasm and in the IMS (Grx1) or in the mitochondrial matrix (Grx2) as a tool for restoring a correct redox environment and preventing the aggregation of mutant SOD1. Here we show that the overexpression of Grx1 increases the solubility of mutant SOD1 in the cytosol but does not inhibit mitochondrial damage and apoptosis induced by mutant SOD1 in neuronal cells (SH-SY5Y) or in immortalized motoneurons (NSC-34). Conversely, the overexpression of Grx2 increases the solubility of mutant SOD1 in mitochondria, interferes with mitochondrial fragmentation by modifying the expression pattern of proteins involved in mitochondrial dynamics, preserves mitochondrial function and strongly protects neuronal cells from apoptosis. The toxicity of mutant SOD1, therefore, mostly arises from mitochondrial dysfunction and rescue of mitochondrial damage may represent a promising therapeutic strategy.

Related Reagents: Parkin Parkin-2 PARK2 Co-regulated (PACRG) PARK7 (DJ-1) LRRK2 (PARK8)                         Neurodegenerative Disease Research Antibodies

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