Ultrastructural studies of the supraoptic nucleus (SON) of the hypothalamus suggest that an active retraction and extension of astrocytic processes (structural plasticity) from between magnocellular neuroendocrine neurons plays a role in the release of oxytocin, vasopressin, or both peptides that accompanies parturition, lactation, and dehydration. In support of this, Salm et al. (1985) previously demonstrated a lactation-associated reduction in immunoreactive glial fibrillary acidic protein (GFAP), an astrocyte-specific cytoskeletal constituent. To determine if similar changes occur in response to dehydration, and if they are reversible, the present study examined GFAP-immunoreactivity (IR) in the SON under various hydration states. Rats were dehydrated for 7 days by substitution of drinking water with 2% saline (n = 3), or dehydrated for 7 days followed by 7 days of rehydration (n = 3). A control group (n = 3) with free access to tap water was used for comparisons. The optical density of GFAP-IR was obtained from the SON, globus pallidus, and lateral hypothalamic regions. The areas of the ventral glial limitans subjacent to the SON (SON-VGL) and of linearly equivalent segments of glial limitans more distant from the SON were also determined. Dehydration resulted in a significant reduction in GFAP-IR in the SON compared to control and rehydrated levels. We also found that the area of the SON-VGL was significantly larger than that of linearly equivalent segments of glial limitans elsewhere and that it was significantly reduced in dehydrated rats, returning to control levels with rehydration. GFAP-IR and glial limitans thickness in regions unrelated to body fluid homeostasis lateral to the SON, overlying to dorsal cortex, and subjacent to the optic chiasm were not significantly changed by hydration state. These results are similar to the changes of GFAP-IR reported for lactating rats and provide further evidence for a role of structural plasticity of astrocytes in events surrounding the selective functional activation of local neurons.