Established immortalized Lymphoblast cell lines from Huntintin patients as well as non-huntingtin patients were used for this study.
Transgenic mice:YAC18, line 29 expressing normal Htt and mutant YAC72, similar to huntingtin disease level of Htt were used. Two types of mutant YAC72, a "high" and "low" level of Htt expression were used. Line 44 has twice the endogenous level (high)and number 2511 has one-half (low) the level of endogenous Htt.
Mitochondria were isolated from lymphoblasts cultured cells and mouse brains.
Mitochondria membrane calcium retention and flux was monitored with tetraphenyl phosphonium (TPP+)-sensitive electrode or calcium green-5V fluorescence. Mitochondria were incubated in medium with sucrose, potassium chloride, glycyl-glycine, potassium phosphate, succinate and calcium chloride.
GST fusion proteins having 19 or 62 poly-L-glutamines were prepared as well as control fusion proteins.
Electron immunocytochemistry used the anti-huntingtin (EM48) antibody which is directed against amino acids 1–256 of human Htt, and selectively labels aggregated Htt26.
Statistical comparisons between two groups made by unpaired t-test and comparisons between more than two groups used ANOVA.