ABSTRACT
TAR DNA-binding protein 43 (TDP-43) has been identified as the major constituent of the proteinaceous inclusions that are characteristic of most forms of amyotrophic lateral sclerosis (ALS) and ubiquitin positive frontotemporal lobar degeneration (FTLD). Wild type TDP-43 inclusions are a pathological hallmark of >95% of patients with sporadic ALS and of the majority of familial ALS cases, and they are also found in a significant proportion of FTLD cases. ALS is the most common form of motor neuron disease, characterized by progressive weakness and muscular wasting, and typically leads to death within a few years of diagnosis. To determine how the translocation and misfolding of TDP-43 contribute to ALS pathogenicity, it is crucial to define the dynamic behavior of this protein within the cellular environment. It is therefore necessary to develop cell models that allow the location of the protein to be defined. We report the use of TDP-43 with a tetracysteine tag for visualization using fluorogenic biarsenical compounds and show that this model displays features of ALS observed in other cell models. We also demonstrate that this labeling procedure enables live-cell imaging of the translocation of the protein from the nucleus into the cytosol.
Subject(s)
Amyotrophic Lateral Sclerosis/metabolism , Cysteine/chemistry , DNA-Binding Proteins/metabolism , Fluoresceins/chemistry , Models, Biological , Organometallic Compounds/chemistry , Sequence Tagged Sites , Amino Acid Sequence , Cell Line, Tumor , Cell Nucleus/metabolism , Cytosol/metabolism , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , Electroporation , Fluorescent Dyes , Frontotemporal Lobar Degeneration/metabolism , Humans , Kinetics , Optical Imaging , Protein Transport , Time-Lapse Imaging , TransfectionABSTRACT
Tubal occlusion using Filshie clip is one of the most commonly performed operations for female sterilization. It is usually a simple and safe procedure, and operative complications are uncommon. We report a rare case of spontaneous expulsion of a Filshie clip through the anterior abdominal wall 5 years after sterilization. The management and possible underlying mechanisms are discussed and the related literature is reviewed.
