Stroke is a major cause of mortality in Portugal and morbidity related to brain lesion in the world. Ischemic stroke occurs when blood flow is interrupted, depleting oxygen and nutrients supply in the tissue. After, blood flow is re-established, the tissue suffers a second damage due to increased oxidative stress. This disease is characterized by excessive oxidative stress, inflammation, excitotoxicity, and, ultimately, cell death. Although therapies are available, they do not target directly brain tissue and present some limitations in efficacy and a short time window of application. Therefore, novel therapeutic approaches against cerebral ischemia are urgently needed. The rational of this thesis is to target glial cells for promoting cerebral protection by limiting neuroinflammation and glial cell death. Central nervous system (CNS) associated disorders are usually considered as a neuronal issue. However, glia are key cells implicated in stress response and involved in neuroprotection; prevention of glial dysfunction strongly protects brain tissue and, ultimately, the patient outcome. Glial cells, such as astrocytes and microglia, are the major cellular group of the brain. These cells are vital players in the brain homeostasis, having roles in development and synaptic plasticity, inflammatory response, metabolic support, and neuronal survival. Astrocytes have important functions in modulation and maturation of synapsis, metabolic support and redox and excitotoxicity protection of neurons. Being in constant microenvironment sensing and in communication with their neighbours, astrocytes are highly sensitive cells able to perform a quicky evaluation of the CNS and quicky intervene to ensure homeostasis and plasticity. On the other hand, microglia are also phagocytic and immunocompetent cell population responsible for the inflammatory response. Although an efficient inflammatory response against infection or injury is crucial to preserve the CNS function; prolonged or exacerbated inflammation is a hallmark of several CNS diseases, namely cerebral ischemia. Therefore, this thesis intends to contribute to the development of approaches (i) that improve astrocyte responsiveness to their microenvironment, and (ii) that limit microglial neuroinflammatory response, which may promote CNS homeostasis, namely against cerebral ischemia. Carbon monoxide (CO) is an endogenous gas that results from haem degradation by the enzyme haem oxygenase. CO is widely described in the literature as an anti neuroinflammatory, an anti-apoptotic and a modulator of cell metabolism molecule. Exposure to low doses of CO or to endogenous CO limits cell death and neuroinflammation in cultures of neurons, astrocytes and microglia, as well as in in vivo models of stroke and neonatal hypoxia ischemia. Nevertheless, CO’s downstream players are not fully described. Disclosing CO’s partners in its protective molecular mechanism will allow a deeper understanding of CO’s biological role, potentially leading to the development of better CO-based therapies, particularly against cerebral ischemia. Therefore, this thesis aims to deeper understand the underlying molecular mechanisms of in CO’s cytoprotection in glia cells. Chapter I introduces main information present in the literature regarding glial cells and CO, as well as the downstream player candidates, Neuroglobin (Ngb) and primary cilia. Chapter II targets the role of Ngb on CO’s anti inflammatory role in microglia, in particular by assessing Ngb relevance in CO’s modulation of cellular metabolism. Chapter III focuses on the ability of CO to modulate the astrocytic primary cilia, which are organelles related to the surveillance of the microenvironment of the cell. These data are then discussed in Chapter IV, where unanswered question and important future work is proposed. Finally, as an annex, Chapter V presents data regarding a technical manuscript in preparation, focused on de novo lipogenesis. This manuscript was developed in collaboration, within the scope of the assessment of CO’s modulation of microglial metabolism.