Determining the Localization of Absent in Melanoma (AIM2) Protein in Human Fibroblast Cells in 2D and 3D Environments
Cells use varying modes of migration depending on their environment. Cells migrating in a highly crosslinked 3D cell-derived matrix (CDM) use a nuclear piston mode of migration. Piston cells form pressure-driven protrusions termed lobopodia to squeeze their bulky nucleus through narrow pores in the CDM. Cells have defenses to prevent damage to DNA and during migration, cells require extra damage control. The protein Absent in Melanoma (AIM2), a cytosolic DNA sensor, is known to mediate DNA damage, but its cellular localization is unknown in migrating cells. AIM2 is also part of the actomyosin machinery that creates lobopodial protrusions. As piston cells migrate through tight spaces, the nucleus briefly ruptures, allowing DNA to leak into the cytosol where AIM2 activates the AIM2 inflammasome to further protect the cell. This work aimed to compare AIM2 localization in 2D and 3D migrating fibrosarcoma cells. We hypothesized that AIM2 localizes perinuclearly in both 2D and 3D migrating cells. We performed an immunofluorescence assay to determine AIM2 localization on 2D glass and in 3D CDM. Results showed AIM2 is located in the nuclear envelope in both 2D and 3D, with more cytoplasmic signal in 2D cells. We concluded that the function of AIM2 is dependent on dimensionality.
Cells use varying modes of migration depending on their environment. Cells migrating in a highly crosslinked 3D cell-derived matrix (CDM) use a nuclear piston mode of migration. Piston cells form pressure-driven protrusions termed lobopodia to squeeze their bulky nucleus through narrow pores in the CDM. Cells have defenses to prevent damage to DNA and during migration, cells require extra damage control. The protein Absent in Melanoma (AIM2), a cytosolic DNA sensor, is known to mediate DNA damage, but its cellular localization is unknown in migrating cells. AIM2 is also part of the actomyosin machinery that creates lobopodial protrusions. As piston cells migrate through tight spaces, the nucleus briefly ruptures, allowing DNA to leak into the cytosol where AIM2 activates the AIM2 inflammasome to further protect the cell. This work aimed to compare AIM2 localization in 2D and 3D migrating fibrosarcoma cells. We hypothesized that AIM2 localizes perinuclearly in both 2D and 3D migrating cells. We performed an immunofluorescence assay to determine AIM2 localization on 2D glass and in 3D CDM. Results showed AIM2 is located in the nuclear envelope in both 2D and 3D, with more cytoplasmic signal in 2D cells. We concluded that the function of AIM2 is dependent on dimensionality.