Digital holographic microscopy discriminates sex differences in medial prefrontal cortex GABA neurons following amphetamine sensitization

Sex differences have been noted in patterns of drug use and relapse, and in particular with amphetamine abuse, implicating estradiol in mediating female neurobehavioral responses. To investigate the interaction of estradiol with amphetamine-induced hyperactivity, we compared male, intact female (INTACT), ovariectomized (OVX) and ovariectomized estradiol-treated (OVX + EB) female rats receiving repeated amphetamine (AMPH) treatment. All rats received intermittent AMPH injections for three days, and baseline and post-injection locomotor activity as well as fine-motor movements were recorded. Upon completion of behavioral experiments, immunohistochemistry was performed to assess parvalbumin-immunoreactive (PV-IR) GABAergic neurons in the medial prefrontal cortex (mPFC). Results indicate that AMPH induced greater behavioral response during habituation among the INTACT animals, and post-injection hyperactivity was apparent on days 2 and 3, among INTACT and OVX + EB females. For INTACT animals, the hyperactivity was most pronounced when estrogen levels were high. Immunohistochemical analysis using digital holographic microscopy revealed INTACT and OVX + EB females had less expression and smaller somatic area of PV-IR neurons in the mPFC. These data provide evidence for rapid development of sex differences in response to AMPH that correlates with sexually dimorphic alterations in a subset of mPFC GABAergic neurons implicated in modulating forebrain dopamine projections.