The sympathetic nervous system has great influence on cardiovascular physiology. Cardiac neurotransmission single photon emission computed tomography (SPECT) imaging allows in vivo noninvasive assessment of presynaptic reuptake and storage of neurotransmitters, which offers characterization of the cardiac neuronal function in different diseases of the heart and other altered metabolic or functional conditions. Therefore assessment of the integrity of cardiac sympathetic innervation may help in the diagnosis of these disorders, as well as in prognostication, and will result in better therapy and outcome. At present, the most widely available SPECT tracer by which to assess cardiac neurotransmission is metaiodobenzylguanidine labeled with iodine 123. This article focuses on reviewing the characteristics of cardiac SPECT imaging with I-123 metaiodobenzylguanidine and its role in the assessment of pathophysiologic changes during relevant clinical conditions.