Fig. 5

Postnatal alcohol exposure (PAE) activates microglia in the cerebellar vermis. Representative images of the cerebellar vermis are stained for ionized calcium-binding adapter protein molecule 1 (IBA-1, red) to label microglia and 4′,6-diamidino-2-phenylindole (DAPI, blue) to label cell nuclei. Microglial morphology was quantified as resting, transitional 1 (T1), transitional 2 (T2), or amoeboid as shown in the top right panel. The proportion of microglia exhibiting each morphology was quantified during the first withdrawal period on P4 (a–b), the third withdrawal period on P6 (d–e), and also on P45 (g–h) (as shown in Fig. 1a). To investigate regional differences, the cerebellar vermis was divided into three lobule regions and the proportion of microglia in an amoeboid morphology was quantified in each region on P4 (c) and P6 (f). On P45 (i), the proportion of resting microglia was quantified by region as there were no amoeboid microglia. (*p < 0.05, **p < 0.01, ***p < 0.001). n = 4 animals from 4 litters. Scale bars are 40 μm and 10 μm for representative low and high magnification images, respectively. The external granule cell (EGL), Purkinje cell (PCL), and internal granule cell (IGL) layers are labeled in panels a and g