Immunolabeling Protocols for Studying Meiosis in Plant Mutants Defective for Nuclear Envelope Components

The nuclear envelope (NE) is a dynamic boundary that allows the communication between nuclear and cytoplasmic components. It has essential roles in a variety of physiological processes including cell division. The linker of nucleoskeleton and cytoskeleton (LINC) complexes span the NE and are important during meiosis, the specialized cell division needed for sexual reproduction. During this division, the LINC complex proteins AtSUN1 and AtSUN2, located in the inner nuclear membrane (INM), are involved in tethering telomeres to the NE. This attachment promotes chromosome movements by the forces that are generated in the cytoplasmic face. In Arabidopsis, the double mutant Atsun1 Atsun2 exhibits a delayed prophase I meiotic progression, partial synapsis, and recombination defects that lead to the formation of unbalanced gametes and sterility. In meiocytes from these mutants, immunolabeling can be applied to analyze possible changes in the dynamics of different meiotic proteins. In addition, if the specific antibodies are available, this technique is an easy and useful tool to determine the spatial distribution of NE proteins.