eCite Digital Repository
Intracellular dialysis disrupts Zn2+ dynamics and enables selective detection of Zn2+ influx in brain slice preparations
Citation
Aiba, I and West, AK and Sheline, CT and Shuttleworth, CW, Intracellular dialysis disrupts Zn2+ dynamics and enables selective detection of Zn2+ influx in brain slice preparations, Journal of Neurochemistry, 125, (6) pp. 822-831. ISSN 0022-3042 (2013) [Refereed Article]
Copyright Statement
2013 International Society for Neurochemistry
Abstract
We examined the impact of intracellular dialysis on fluorescence detection of neuronal intracellular Zn2+ accumulation. Comparison between two dialysis conditions (standard; 20 min, brief; 2 min) by standard whole-cell clamp revealed a high vulnerability of intracellular Zn2+ buffers to intracellular dialysis. Thus, low concentrations of zinc-pyrithione generated robust responses in neurons with standard dialysis, but signals were smaller in neurons with short dialysis. Release from oxidation-sensitive Zn2+ pools was reduced by standard dialysis, when compared with responses in neurons with brief dialysis. The dialysis effects were partly reversed by inclusion of recombinant metallothionein-3 in the dialysis solution. These findings suggested that extensive dialysis could be exploited for selective detection of transmembrane Zn2+ influx. Different dialysis conditions were then used to probe responses to synaptic stimulation. Under standard dialysis conditions, synaptic stimuli generated significant FluoZin-3 signals in wild-type (WT) preparations, but responses were almost absent in preparations lacking vesicular Zn2+ (ZnT3-KO). In contrast, under brief dialysis conditions, intracellular Zn2+ transients were very similar in WT and ZnT3-KO preparations. This suggests that both intracellular release and transmembrane flux can contribute to intracellular Zn2+ accumulation after synaptic stimulation. These results demonstrate significant confounds and potential use of intracellular dialysis to investigate intracellular Zn2+ accumulation mechanisms.
Item Details
Item Type: | Refereed Article |
---|---|
Keywords: | hippocampal slice, metallothionein, pyrithione, whole-cell, Zinc, ZnT3 |
Research Division: | Biomedical and Clinical Sciences |
Research Group: | Neurosciences |
Research Field: | Central nervous system |
Objective Division: | Health |
Objective Group: | Clinical health |
Objective Field: | Clinical health not elsewhere classified |
UTAS Author: | West, AK (Professor Adrian West) |
ID Code: | 85242 |
Year Published: | 2013 |
Web of Science® Times Cited: | 7 |
Deposited By: | Menzies Institute for Medical Research |
Deposited On: | 2013-06-20 |
Last Modified: | 2017-11-06 |
Downloads: | 0 |
Repository Staff Only: item control page