We previously showed that prothymosin alpha (ProT alpha) improves cerebral ischemia-induced motor dysfunction. Our recent study also demonstrated that heterozygous ProT alpha deletion exhibited an enhanced anxiety-like behavior in mice. However, it remains elusive which brain regions or cells are related to these phenotypes. Here we generated conditional G gamma 7-specific ProT alpha knockout mice using G protein gamma 7 subunit gene (Gng7)-cre promoter to see the brain robustness roles of ProT alpha in the striatum and hippocampus. The younger conditional ProT alpha (Gng7) knockout mice at the age of 10 weeks showed no significant phenotypes in motor dysfunction in the Rotarod test and locomotor activity in the open-field test, whereas significant motor dysfunction was obtained by 15 min transient middle cerebral artery occlusion (tMCAO)-induced cerebral ischemia. The aged conditional ProT alpha (Gng7) knockout mice at the age of 20 weeks showed hypolocomotor activity with less center time in the open-field test and impaired motor coordination in the Rotarod test without ischemia. Thus, this study suggests that ProT alpha has important roles in the maintenance of motor coordination and anxiety-like behavior. (C) 2019 Elsevier Inc. All rights reserved.