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Objective Dopamine is a neurotransmitter that plays crucial role in the movement, reward and emotional response.Neurons containing dopamine are mainly clustered in mesencephalon.These dopaminergic (DA) neurons formed nigrostriatal, mesocortical and mesolimbic projections.Mesocortical projection is closely associated with psychiatric disorder.However, the molecular mechanisms which specify mesocortical projection, remain elusive.Recent studies showed that Lim homeobox transcription factor 1-beta (lmxlb) involved in the development of the midbrain.Methods To address the role of limlb in mesocortical projection, we injected retrograde fluorescent microspheres into the prefrontal cortex of neonatal mice and labeled mesocortical DA subtype neurons.Additionally, immunohistochemical staining was performed to examine the expression of lmx1b, and in situ hybridization was further performed to examine the signal of limx1b in mesencephalon.Results Mesocortical neurons were labeled with injection fluorescent microspheres in prefrontal cortex.Immunohistochemical staining demonstrated that lirnx 1 b was expressed in labeled cells.Moreover, Limx1b could be detected in mesencephalon.Conclusion This study showed that lmx1b involved mesocortical projection, which is significant for understanding the pathogenesis of psychiatric disorder.