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Automated three-dimensional reconstruction of the Caenorhabditis elegans germline

Gopal, Sandeep, Boag, Peter, Pocock, Roger
Developmental biology 2017
Caenorhabditis elegans, apoptosis, chromosomes, cytoskeleton, germ cells, methodology, models, mutation, phenotype, protein synthesis, proteins, stem cells
The Caenorhabditis elegans germline is widely used as a model to study stem cell development, chromosome dynamics and apoptosis. Major readouts of germline phenotypes such as cell counting and protein expression profiling are routinely analysed manually and in a two-dimensional manner. The major disadvantages of the existing approaches are 1) they are time-consuming and laborious and 2) there is an inability to study the effects of genetic mutations in three dimensions. Here, we demonstrate a rapid, automated method for analysing the three-dimensional distribution of proteins, germline nuclei and cytoskeletal structures in the C. elegans germline. Using this method, we have revealed previously unappreciated germline organisation and cytoskeletal structures that will have a major impact on the characterisation of germline phenotypes. To conclude, our new method dramatically enhances the efficiency and resolution of C. elegans germline analysis and may be applied to other cellular structures.