ИСТИНА

Progress in fundamental research is directly related to the emergence of new research methods that not only expand the established classical concepts, but can provide information that fundamentally changes them. . According to our data, vimentin filaments, binding to mitochondria, determine their distribution and mobility in cells (Nekrasova et al., 2011), affect the level of their membrane potential (Chernoivanenko et al., 2015), there is a region responsible for the interaction of vimentin filaments with mitochondria in the N-terminal part of the vimentin molecule (Nekrasova et al., 2011) and similar amino acid sequences are found in other proteins, for example, in desmin. Since direct interaction of vimentin filaments with microtubules (Schaedel et al., 2021) and actin filaments (Esue et al., 2006) has already been shown, these facts together allowed us to assume that the connection of individual cytoskeleton components with each other and with mitochondria is not limited to interaction through cross-linker and motor proteins. Vimentin (and possibly other intermediate filament proteins) can regulate cytoskeletal interactions with mitochondria (Alieva et al., 2024). A revolutionary study performed using cryo-electron tomography and fundamentally changing our understanding of the three-dimensional structure of vimentin filaments (Eibauer et al., 2024) motivated us to use the capabilities of the cryo-electron microscopy method to try to identify the binding sites of vimentin with other cytoskeletal components and mitochondria. To meet this challenge is fundamentally possible if we combine super-resolution microscopy (3D-SIM) and cryo-electron tomography. The task is to identify and to characterize in detail the ultrastructure of the binding sites of vimentin (and possibly other intermediate filaments) to mitochondria - in particular, to test whether the N-terminus of vimentin (desmin) extends through the outer mitochondrial membrane into the intermembrane space, or whether it penetrates through two membranes and enters the mitochondrial matrix Historically, the main problem for addressing appropriate biological questions for fluorescence correlative light and electron microscopy has been the so-called ‘resolution gap’. We hope that 3D-SIM usage instead of conventional fluorescence microscopy in combination with cryo-electron tomography will allow us to overcome the ‘resolution gap’ and solve our problems.