A number of recent discoveries suggest close links between anxiety- or probable relief-like behaviors and intracellular signaling molecules controlling morphogenesis in glial cells such as oligodendrocytes (oligodendroglial cells) in the brain, which have protective effects on neuronal cells. In the former behaviors, their intracellular signaling molecules include small GTPase members, some of which mediate cell morphological changes. Rnd2 is one such member, belonging to the Rho family of small GTPases. Despite the currently known functions of Rnd2, the precise roles of Rnd2 in cell morphogenesis and related functions in health and disease states remain to be elucidated. Herein we show that signaling through anxiety-related loss of function of the rnd2 gene is related to the regulation of oligodendroglial cell morphological differentiation in the FBD-102b cell line, which is often utilized as oligodendroglial cell differentiation model. Knockdown of Rnd2 with the clustered regularly interspaced palindromic repeats (CRISPR)/CasRx system or RNA interference has been shown to inhibit morphological differentiation. Similarly, knockdown of Prag1 and Fyn kinase, signaling molecules acting downstream of Rnd2, also blunts differentiation. Rnd2 or Prag1 knockdown also decreases Fyn phosphorylation, which is critical for its activation and for oligodendroglial cell differentiation and myelination. Of note, hesperetin, a citrus flavonoid with protective effects on oligodendroglial cells as well as neuronal ones, can recover the defective differentiation induced by the knockdown of Rnd2/Prag1/Fyn. These results suggest that signaling through Rnd2/Prag1/Fyn is directly associated with normal oligodendroglial cell morphological differentiation. Deficiency of the signaling cascade is recovered by hesperetin, presenting one potential molecular construction underlying anxiety and possible therapeutic targets.