MECHANICALLY INDUCED REACTIVE GLIOSIS CAUSES ATP-MEDIATED ALTERATIONS IN ASTROCYTE STIFFNESS

Abstract

Reactive gliosis is a process triggered in astrocytes after traumatic injury, yet the exact consequences of gliosis on cellular survival and neural regenerative processes in the injured brain remain only partly understood. One recently discovered feature influencing neuronal growth and differentiation is the physical stiffness of the environment surrounding pioneering neurites. In this study, the mechanical properties of cultured cortical astrocytes are measured following a mechanical stretch injury that induces reactive gliosis. In mechanically injured cultures, there was a significant increase in glial fibrillary acidic protein (GFAP) immunoreactivity 24 h following a rapid, transient 15% strain. In these same cultures, astrocytes in the surrounding region--the "mechanical penumbra"--also exhibited increased GFAP immunoreactivity compared to naive cultures. Correlated with these changes in GFAP was a general softening of the non-nuclear regions of the astrocytes, both in the injured and penumbra cells, as measured by atomic force microscopy (AFM). The elastic modulus in naive cultures was observed to be 57.7+/-5.8 kPa in non-nuclear regions of naive cultures, while 24 h after injury the modulus was observed to be 26.4+/-4.9 kPa in the same region of injured cells. In the penumbra of injured cultures, the modulus was 23.7+/-3.6 kPa. Alterations in astrocyte stiffness in the area of injury and mechanical penumbra were ameliorated by pretreating cultures with a nonselective P2 receptor antagonist (PPADS). Since neuronal cells generally prefer softer substrates for growth and neurite extension, these findings may indicate that the mechanical characteristics of reactive astrocytes are favorable for neuronal recovery after traumatic brain injury.