The mechanisms and time-course of change in neuromuscular function after intracellular dehydration and subsequent rehydration, are not well understood. In this present study, twelve healthy participants underwent a control trial (CON) and two experimental trials in heat (45 °C, 45% relative humidity [RH]) to achieve a 5% reduction in body mass via dehydration and low-intensity cycling (60 W), then rehydrated rapidly (RHY2) or progressively (RHY24). Participants underwent various measures of transcranial magnetic stimulation (TMS) and peripheral motor nerve stimulation (MNS) before (PRE), after core temperature (CT) returned to baseline (POST), and after 2 h and 24 h. These measures included: motor evoked potential amplitude (MEP), TMS-evoked cortical silent period (cSP), compound muscle action potential (MMAX), voluntary activation (VA), potentiated twitch (Qtw,pot), maximal voluntary contractions (MVC) and serum Ubiquitin carboxyl-terminal hydrolase (UCH-L1). The novel finding was a different corticospinal response to low-intensity exercise (i.e., higher corticospinal inhibition when hydrated) and rehydration strategies (i.e., lower corticospinal excitability after gradual rehydration).