Reactive astrogliosis propagation in a model of in vitro scratch injury

CIERI, MARÍA BELÉN; MURTA, VERÓNICA; ROSCISZEWSKI, GERARDO; CADENA, VANESA ; LUKIN, JERÓNIMO; RAMOS, ALBERTO; AUZMENDI, JERÓNIMO

CABA

Congreso; 2nd FALAN CONGRESS |FALAN-IBROLARC; 2016

FALAN

Glial scar formation is a complex process whichinvolve establishment of a physical barrier surrounding of the brain injury. Reactiveastrocytes extend their process near to injury and contribute to form the barrier.In this work we analyze the role of the toll like receptor (TLR), Ca+2,connexin and cell composition over the spread of astrogliosisand the glial scarformation. Using an in vitro woundmodel, first we showed that TLR2 and TLR4 increasing 1.3 times the astrocyticinvasive capability with a different dynamic profile in an NFκB dependentmanner.  On the other hand, extracellularCa+2 is required to trigger the invasion in connexin independentmode. Furthermore, stimulating TLRs ways, blocking connexin or chelating Ca+2,we find an increase of 10-20% in GFAP expression with a gradient expression profilefrom the injury. Additionally, astrocytes cultured without microglia showed asimilar level of GFAP expression, but only TLR2 way keep the same expressionprofile.  Last, to evaluate thecontribution of TLRs ways to scar formation we use the same experimental modelover an aged mixed glial culture. Only the stimulation of TLR4 with LPSgenerated a structure like scar at 24h after injury. We conclude that TLRs waysplays a key role in the spread of the astrogliosis in a NFκB dependent manner butthe glial scar formation require a more complex environment where other celltype are necessary.