Traumatically injured astrocytes release a proteomic signature modulated by STAT3-dependent cell survival

Molecular markers associated with CNS injury are of diagnostic interest. Mechanical trauma generates cellular deformation associated with membrane permeability with unknown molecular consequences. We used an in vitro model of stretch‐injury and proteomic analyses to determine protein changes in murine astrocytes and their surrounding fluids. Abrupt pressure‐pulse stretching resulted in the rapid release of 59 astrocytic proteins with profiles reflecting cell injury and cell death, i.e., mechanoporation and cell lysis. This acute trauma‐release proteome was overrepresented with metabolic proteins compared with the uninjured cellular proteome, bearing relevance for post‐traumatic metabolic depression. Astrocyte‐specific deletion of signal transducer and activator of transcription 3 (STAT3‐CKO) resulted in reduced stretch‐injury tolerance, elevated necrosis and increased protein release. Consistent with more lysed cells, more protein complexes, nuclear and transport proteins were released from STAT3‐CKO versus nontransgenic astrocytes. STAT3‐CKO astrocytes had reduced basal expression of GFAP, lactate dehydrogenase B (LDHB), aldolase C (ALDOC), and astrocytic phosphoprotein 15 (PEA15), and elevated levels of tropomyosin (TPM4) and α actinin 4 (ACTN4). Stretching caused STAT3‐dependent cellular depletion of PEA15 and GFAP, and its filament disassembly in subpopulations of injured astrocytes. PEA15 and ALDOC signals were low in injured astrocytes acutely after mouse spinal cord crush injury and were robustly expressed in reactive astrocytes 1 day postinjury. In contrast, α crystallin (CRYAB) was present in acutely injured astrocytes, and absent from uninjured and reactive astrocytes, demonstrating novel marker differences among postinjury astrocytes. These findings reveal a proteomic signature of traumatically‐injured astrocytes reflecting STAT3‐dependent cellular survival with potential diagnostic value. GLIA 2016;64:668–694 Main points Mechanically traumatized astrocytes leak proteins into surrounding fluids. The fluid derived astroglial trauma‐release proteome captures profiles of astrocyte injury and cell death. STAT3 regulates cell survival based on the energy capacity of injured astrocytes.