The experimental analysis of the neuroendocrine interactions regulating sexual behavior has traditionally relied on studying the effects of CNS lesions and pharmacological treatments with hormones or drugs purportedly acting through specific neurotransmitter systems. New methodological developments have allowed the assessment of several indices of neural function in experimental animals, particularly the rat, as they relate to behavioral changes. In the field of sexual behavior, ex vivo analyses have been used to measure markers of energy metabolism, such as 2-deoxyglucose uptake and Na,K-ATPase activity, the tissue content of neurotransmitters and metabolites, the levels of steroid receptors and neurosteroids, and immediate-early gene expression products in different areas of the CNS. In vivo studies have monitored brain electrical activity and temperature, as well as the extracellular levels of neurotransmitters and metabolites by cerebrospinal fluid sampling, push-pull perfusion and, especially, electrochemical recordings and microdialysis, in the course of mating and exposure to various relevant stimuli. The findings with the different methodologies are generally consistent and agree with those of previous surgical and pharmacological manipulations. They provide data on temporal relationships between neurobiological and behavioral events and suggest new interpretations for different aspects of the male copulatory pattern.