In order to establish the involvement of particular neurochemical brain groups in the response to blood volume expansion, we analyzed Fos-labeling in combination with immunolabeling for serotonin, tyrosine hydroxylase, vasopressin and oxytocin, 90 min after a sham or i.v. isotonic blood volume expansion (BVE) in unanesthetized, unrestrained rats. We also examined the changes in concentration of oxytocin, atrial natriuretic peptide and vasopressin plasma, induced by blood volume load, to confirm our previous studies. The results demonstrate the participation of specific paraventricular and supraoptic nucleus groups of cells (oxytocinergic-vasopressinergic), serotoninergic dorsal raphe nucleus cells and catecholaminergic A1/A2/A6 groups (in the caudal ventrolateral medulla, nucleus of the solitary tract and locus coeruleus respectively), in the regulatory response to BVE. They provide detailed neuroanatomical evidence to support previous observations showing the contribution of these neurochemical systems in the neural, behavioral and endocrine response to isotonic BVE.