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Highlights • β 3 -receptor protein expression is detectable in rat mPFC tissue. • β 3 -receptors are found in layer V mPFC pyramidal neurons. • Stimulation of β 3 -adrenoreceptors evokes inward currents in mPFC pyramidal neurons. Abstract While the expression of β 3 -adrenergic receptors is firmly established in adipose, kidney and heart tissue, their expression and function in the brain remains unclear despite their potential role in depression and stress-related disorders. This study aimed to investigate the expression of β 3 -adrenoreceptors and their involvement in the mechanism controlling the resting holding current in layer V medial prefrontal cortex (mPFC) pyramidal neurons in young rats. Applications of the selective β 3 -adrenergic receptor agonists BRL 37344 and SR 58611 A evoked inward currents in the tested neurons. The inward currents evoked by BRL 37344 or noradrenaline (a nonselective physiological adrenergic receptor agonist) were prevented or decreased, respectively, by the selective β 3 -receptor antagonist L-748,337. Western blot and fluorescence immunohistochemistry analyses revealed β 3 -adrenergic receptor protein expression in the mPFC. Thus, based on the results obtained here, functional β 3 -adrenergic receptors are expressed in layer V mPFC pyramidal neurons and their activation evokes inward currents. [ABSTRACT FROM AUTHOR]