Figure 2
Sera from permanent middle cerebral artery occlusion (pMCAO) treated gal-3+/+ but not gal-3−/−, mice cause myenteric neuronal loss in vitro.
(A,B) Gal-3+/+ (black circles) and Gal-3−/− (black squares) sera from 3 (A) and 7 days (B) sham mice do not affect myenteric neuronal survival in C57BL/6 derived cultures. Neither do 3 (A) or 7 days (B) pMCAO sera from gal-3−/− mice (open squares), while 3 (A) and 7 days (B) pMCAO sera from gal-3+/+ mice (open circles) significantly reduce myenteric neuronal survival in C57BL/6 derived cultures. (C) Purified gal-3 (10−6 M) induces myenteric neuronal loss in C57BL/6 derived cultures. This was prevented by the presence of inhibitors of TAK1 (TAK1i, 10−6 M) or AMPK (AMPKi, 10−5 M). (D) The gal-3+/+ pMCAO serum (1:100, 3 and 7 days post surgery) -induced neuronal loss could be prevented by presence of inhibitors of TAK1 (TAK1i, 10−6 M) or AMPK (AMPKi, 10−5 M). (E) Neither LPS nor sera (1:200) from sham or pMCAO 3 and 7 days treated gal-3+/+ and gal-3−/− mice affect survival of myenteric neuronal in TLR4−/− derived cultures. (F) Western blot analysis of gal-3 protein levels in immune cells collected by peritoneal lavage and digestive organs. Results are expressed as % neuronal survival compared to controls run in parallel (A–E) or as % gal-3 of actin (F), in mean ± SEM, n = 3–18, * P < 0.05, **P < 0.01, ***P < 0.001.
