Acute and chronic stress can induce the production of reactive oxygen species, which exert a multitude of biological effects covering a wide spectrum, that ranges from physiological regulatory functions to damaging alterations participating in the pathogenesis of increasing number of diseases. With the aim of elucidating the impact of acute and chronic stress on oxidative and antioxidative response in nuchal muscles (Semispinalis capitis, Splenius capitis, Splenius cervicis) of male (M), female (NE-OV) and ovariectomized female (OV), Sprague-Dawley rats were exposed to acute and chronic stress protocol. Chronic stress caused oxidative stress, that was determined by the increased lipid peroxidation level (LPO) and decreased antioxidative defense mechanism in nuchal muscles of males and females. As an indicators of antioxidative response, the ratio of reduced and oxidised glutathione (GSH/GSSG), the activity of antioxidative enzymes (catalase - CAT, glutathione-peroxidase - GPX, glutathione-S-transferase - GST, superoxid-dismutase - SOD and glutathione-reductase - GR) were measured. Acute stress did not have significant impact on the most of antioxidative enzyme activities (CAT, GPX, GST, SOD, GR) in males and females, although GSH/GSSG ratio was reduced. Ovariectomy increased LPO and reduced some of antioxidative enzyme activities (CAT, GPX). Oxidative and antioxidative response to acute and chronic stress, in nuchal muscles of males and females, showed similarity in most of the measured parameters, and it differed from the response in ovariectomized females. Unlike acute stress, chronic stress disrupts antioxidative defense signficantly more, thus it can cause changes in muscle contraction and muscle fatigue, which may lead to chronic pain in neck and headache.