Document Detail


Enhanced cystatin C and lysosomal protease expression following 6-hydroxydopamine exposure.
MedLine Citation:
PMID:  16414118     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
6-Hydroxydopamine (6-OHDA) is a selective neurotoxin used to induce apoptosis in catecholamine-containing neurons. Although biochemical products and reactive oxygen species (ROS) of 6-OHDA have been well documented, the activation of cellular pathways following exposure are not well understood. Apoptosis in PC12 (Pheochromocytoma) cells was induced by 6-OHDA in a dose (10-150 microM) and time-dependent (24-72 h) manner compared to experimental controls (no treatment). PC 12 cells exposed to 50 microM 6-OHDA demonstrated the involvement of caspase 3 and lysosomal protease alterations. Following 6-OHDA exposure, the caspase 3-like inhibitor Ac-DEVD-CHO significantly decreased 6-OHDA induced cell death. In addition, alterations in expression of the lysosomal cysteine and aspartic proteases, cathepsin B (CB) and cathepsin D (CD) and the endogenous cysteine protease inhibitor cystatin C were observed utilizing immunocytochemical analysis at 24, 48, and 72 h following 6-OHDA exposure. Furthermore, CB and CD and cystatin C immuno-like reactivity was more pronounced in TUNEL positive cells. Moreover, Western blot analysis confirmed a significant increase in protein expression for CB and CD at 72 h and a temporal and concentration dependent increase in cystatin C in response to 6-OHDA. Cells treated with pepstatin A, an inhibitor for CD, showed a significant decrease in cell death, however, CA-074ME, a specific inhibitor for CB, failed to protect cells from 6-OHDA induced cell death. Thus, these results suggest that apoptosis induced by 6-OHDA exposure is mediated in part through caspase 3 activation and lysosomal protease CD.
Authors:
Daniel C Lee; Fran T Close; Carl B Goodman; Inneke M Jackson; Ceceile Wight-Mason; Lateesha M Wells; Tracy A Womble; Donald E Palm
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2006-01-18
Journal Detail:
Title:  Neurotoxicology     Volume:  27     ISSN:  0161-813X     ISO Abbreviation:  Neurotoxicology     Publication Date:  2006 Mar 
Date Detail:
Created Date:  2006-02-13     Completed Date:  2006-05-22     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  7905589     Medline TA:  Neurotoxicology     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  260-76     Citation Subset:  IM    
Affiliation:
College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, FL 32307, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis / drug effects
Blotting, Western
Caspase 3
Caspases / metabolism
Cathepsin B / biosynthesis
Cathepsin D / biosynthesis
Cell Death / drug effects
Cell Proliferation / drug effects
Cell Survival / drug effects
Cystatin C
Cystatins / biosynthesis*
Cytochromes c / metabolism
DNA / biosynthesis,  genetics
Humans
Immunohistochemistry
In Situ Nick-End Labeling
L-Lactate Dehydrogenase / metabolism
Lysosomes / drug effects,  enzymology*
Oxidative Stress / drug effects
Oxidopamine / toxicity*
PC12 Cells
Peptide Hydrolases / biosynthesis*
Poly(ADP-ribose) Polymerases / metabolism
Rats
Stimulation, Chemical
Sympatholytics / toxicity*
Grant Support
ID/Acronym/Agency:
G12RR03020/RR/NCRR NIH HHS; S06 GM 0811/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/CST3 protein, human; 0/Cst3 protein, rat; 0/Cystatin C; 0/Cystatins; 0/Sympatholytics; 1199-18-4/Oxidopamine; 9007-43-6/Cytochromes c; 9007-49-2/DNA; EC 1.1.1.27/L-Lactate Dehydrogenase; EC 2.4.2.30/Poly(ADP-ribose) Polymerases; EC 3.4.-/Peptide Hydrolases; EC 3.4.22.-/CASP3 protein, human; EC 3.4.22.-/Casp3 protein, rat; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases; EC 3.4.22.1/Cathepsin B; EC 3.4.23.5/Cathepsin D

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