Background: Restrictions due to COVID-19 lockdowns reduced the possibilities of children and adolescents for being active, with negative consequences in adopting a healthy lifestyle. Purpose: To compare Greek adolescents’ self-reported weekly participation in physical activity, during and before the two initial strict lockdowns, due to COVID-19. Secondary aims were to examine these differences with regard to gender, and associations between weekly physical activity participation with health status variables. Methods: Three hundred and sixty-three adolescents (Ν=363) from secondary schools, in the Greek territory (108 boys and 255 girls) filled in the Godin-Shephard Leisure-Time Physical Activity Questionnaire and the TNO-AZL Questionnaire for Children’s Health-Related Quality of Life Children’s Form, online. It was a cross-sectional study and data were collected during first and second strict lockdowns, from different adolescents who filled in the above online questionnaires once. Results: Adolescents reduced significantly weekly frequency of strenuous, moderate and total physical activity, during lockdowns (p.05). Time factor had a different effect on boys and girls, only in moderate physical activity (p.05). Positive correlations were found between strenuous and total physical activity with positive moods, and moderate physical activity with cognitive functioning and school performance, during lockdowns (p.05). Conclusions: This study adds information regarding the negative impact on physical activity participation in Greek adolescents during COVID-19 strict lockdowns. The findings, also, highlighted that reduced physical activity among adolescents during lockdowns associated with poorer psychosocial status. Thus, these results could be used to inform strategies for promoting health movement behaviors, in order to reduce possible negative consequences during future pandemics.