Itch sensation is one of the major sensory experiences of humans and animals. Recent studies using genetic deletion techniques have proposed that gastrin-releasing peptide (GRP) is a key neurotransmitter for itch in the spinal cord. However, these studies are mainly based on behavioral responses and lack direct electrophysiological evidence that GRP indeed mediates itch information between primary afferent fibers and spinal dorsal horn neurons. In this review, we review recent studies using different experimental approaches and propose that glutamate but not GRP acts as the key neurotransmitter in the primary afferents in the transmission of itch. GRP is more likely to serve as an itch-related neuromodulator. In the cerebral cortex, we propose that the anterior cingulate cortex (ACC) plays a significant role in both itch and pain sensations. Only behavioral measurement of itch (scratching) is not sufficient for itch measurement, since scratching the itching area also produces pleasure. Integrative experimental approaches as well as better behavioral scoring models are needed to help to understand the neuronal mechanism of itch and aid future treatment for patients with pruritic diseases.