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Although small GTPases of the Rho family are powerful regulators of cellular morphology, the signaling modules of Rho\u2010mediated pathways in astrocytes remain enigmatic. Here we demonstrated that the serotonin receptor 4 (5\u2010HT<jats:sub>4<\/jats:sub>R) is expressed in hippocampal astrocytes, both in vitro and in vivo. Through fluorescence microscopy, we established that 5\u2010HT<jats:sub>4<\/jats:sub>R activation triggered RhoA activity via G\u03b1<jats:sub>13<\/jats:sub>\u2010mediated signaling, which boosted filamentous actin assembly, leading to morphological changes in hippocampal astrocytes. We investigated the effects of these 5\u2010HT<jats:sub>4<\/jats:sub>R\u2010mediated changes in mixed cultures and in acute slices, in which 5\u2010HT<jats:sub>4<\/jats:sub>R was expressed exclusively in astrocytes. In both systems, 5\u2010HT<jats:sub>4<\/jats:sub>R\u2010RhoA signaling changed glutamatergic synaptic transmission: It increased the frequency of miniature excitatory postsynaptic currents (mEPSCs) in mixed cultures and reduced the paired\u2010pulse\u2010ratio (PPR) of field excitatory postsynaptic potentials (fEPSPs) in acute slices. 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