Determination of copper poisoning in Wilson's disease using laser ablation inductively coupled plasma mass spectrometry.

Abstract

Copper (Cu) is an essential trace element that is vital to the health of all living organisms. As a transition metal, it is involved in a myriad of biological processes. Balance studies estimated that the adult human requirement for copper is in the range of 1.3 to 2 mg per day. Cu deficiency alters immune function, neuropeptide synthesis and antioxidant defense, while the excess in Cu results in oxidative stress and progressive structural damage of mitochondrial and clinically in hepatic and/or neurological symptoms. This becomes particularly visible in Wilson's disease (WD) representing a rare autosomal recessive inherited disorder with a disease prevalence of about 1 in 30,000 people. The affected gene, i.e., ATP7B, belongs to the class of ATP-dependent, P-type Cu-transporting ATPases. To understand the pathomechanism in WD, several experimental models for studying WD were established. Independent studies performed in these models showed that the inactivation of the Atp7b gene results in a gradual increase in Cu in many organs during life span. However, the exact distribution of Cu and the potential impact of elevated Cu concentrations on other metals within the tissue are only sparely analyzed. Recently, novel laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS)-based protocols for metal bio-imaging in liver and brain were established. In the present review, we will discuss the methodological background of this innovative technique and summarize our experiences using LA-ICP-MS imaging in biological monitoring, exact measurement, and spatial assignment of metals within tissue obtained from Atp7b null mice and clinical specimens taken from patients suffering from genetically confirmed WD. Using WD as an example, the data discussed demonstrates that LA-ICP-MS has multi-element capability, allowing precise measurement and visualization of metals in the tissue with high spatial resolution, sensitivity, quantification ability, and exceptional reproducibility.