A central aim in fault mechanics is to understand the microphysical mechanisms controlling aseismic-seismic transitions in fault gouges, and to identify microstructural indicators for such transitions. We present new data on the slip stability of calcite fault gouges, and on microstructural development down to the nanometer scale. Our experiments consisted of direct shear tests performed dry at slip rates of 0.1–10 µm/s, at a constant normal stress of 50 MPa, at 18–150 °C. The results show a transition from stable to (potentially) unstable slip above ∼80 °C. All samples recovered showed an optical microstructure characterized by narrow, 30–40-µm-wide, Riedel and boundary shear bands marked by extreme grain comminution, and a crystallographic preferred orientation (CPO). Boundary shear bands, sectioned using FIB-SEM (focused ion beam scanning electron microscopy), revealed angular grain fragments decreasing from 10 to 20 µm at the outer margins to ∼0.3 µm in the shear band core, where dense aggregates of nanograins also occurred. Transmission electron microscopy, applied to foils extracted from boundary shears using FIB-SEM, combined with the optical CPO, showed that these aggregates consist of calcite nanocrystals, 5–20 nm in size, with the (104) dislocation glide system oriented parallel to the shear plane and direction. Our results suggest that the mechanisms controlling slip include cataclasis and localized crystal plasticity. Because crystal plasticity is strongly thermally activated, we infer that the transition to velocity-weakening slip is likely due to enhanced crystal plasticity at >80 °C. This implies that tectonically active limestone terrains will tend to be particularly prone to shallow-focus seismicity.