Figure 5

Increased phosphorylation of Akt and GSK3β in neuronal cells of sonicated rat brain regions. Paraffin sections of rat brains were immunofluorescently stained with primary antibodies against astrocyte marker, Glial Fibrillary Protein (GFAP, green), Neuronal marker (NeuN, green), phospho-Akt (pAkt, red) and phospho-GSK3 β (pGSK3β, red). In all sections, DAPI (blue) represents nuclear staining of the cells. A, Panel [i] shows a control section without application of any primary antibodies. Panel [ii] represents a direct staining of the astrocytes with Alexa-fluor488-conjugated GFAP (green, right arrow). Panels [iii] and [iv] represent immunofluorescence staining with pAkt (red, curved arrow) and GFAP (green, right arrow) in non-sonicated (-US) as well as sonicated (+US) hemispheres respectively. Panels [v] and [vi] illustrate co-staining of pGSK3β (red, pentagon) and GFAP (green, right arrow) in '-US' and '+US' brain regions respectively. B, Panels [ii] and [iii] represent neuronal cells morphology as directly stained with Alexa-Fluor 488-conjugated NeuN (green, lightning bolt). Co-staining of pAkt (red, curved arrow) with NeuN (green) and also pGSK3β (red, pentagon) with NeuN (green) are shown in panels [iii, iv] and [v, vi] respectively. The regions of the brain sections with IgG extravasation (arrow head) and their surrounding neuronal cells with elevated levels of pAkt and pGSK3β are shown in panels [iv, vi].