Fig. 7

Reactive astrogliosis in brainstem of mTBI mice 7 days following injury. a An example brainstem section at the Bregma level -8.2 stained with GFAP (green, immunohistochemistry) and Gfap (red, in situ hybridization). Scale bar = 200 µm. b Insets in Fig. 7a representing differences in Gfap mRNA/GFAP antibody (Ab) staining in various brainstem regions. Representative confocal images from a sham (c) and a mTBI (d) mouse showing colocalization of Gfap mRNA (red, in situ hybridization) on GFAP + astrocytes (green, immunohistochemistry) in the Sp5c. Scale bar = 50 µm. Inset of 7d from a mTBI mouse Sp5C shows increased Gfap mRNA expression in the processes of GFAP + astrocytes in lamina III of Sp5C. Scale bar = 20 µm. e) Graph representing percent Gfap mRNA expression in GFAP + fibrous astrocytes in the Sp5c of sham (n = 155 fibers) and mTBI (n = 168 fibers) groups. f) Close proximity of GFAP-ir fibers or cells to Iba1/CD68 + cells in different regions of the brainstem are shown in white arrows. GFAP-ir fibers or cells that are not in close contact with Iba1/CD68 + cells are shown in purple arrows. g) Representative images showing localized expression of Slc1a2 in the Sp5C of sham and mTBI mice. Scale bar = 500 µm. h) Presence of GFAP + astrocytes (green, immunohistochemistry) in close proximity to Slc1a2 mRNA puncta (purple, in situ hybridization) in the lamina III of Sp5C are shown in white arrows. Scale bar = 50 µm. IRt—Intermediate reticular nucleus; Mdv—Medullary reticular nucleus, ventral part; py—pyramidal tract; sc—spinocerebellar tract; Sp5c (MDH)- spinal trigeminal nucleus caudalis (medullary dorsal horn); ts—tectospinal tract