Orthostatic testing, involving the transition from different body positions (e.g., from lying or sitting position to an upright or standing position), offers valuable insights into the autonomic nervous system (ANS) functioning and cardiovascular regulation reflected through complex adjustments in, e.g., measures of heart rate (HR) and heart rate variability (HRV). This narrative review explores the intricate physiological mechanisms underlying orthostatic stress responses and evaluates its significance for exercise science and sports practice. Into this matter, active orthostatic testing (e.g., active standing up) challenges the cardiovascular autonomic function in a different way than a passive tilt test. It is well documented that there is a transient reduction in blood pressure while standing up, leading to a reflex increase in HR and peripheral vasoconstriction. After that acute response systolic and diastolic blood pressures are usually slightly increased compared to supine lying body position. The ANS response to standing is initiated by instantaneous cardiac vagal withdrawal, followed by sympathetic activation and vagal reactivation over the first 25-30 heartbeats. Thus, HR increases immediately upon standing, peaking after 15-20 beats, and is less marked during passive tilting due to the lack of muscular activity. Standing also decreases vagally related HRV indices compared to the supine position. In overtrained endurance athletes, both parasympathetic and sympathetic activity are attenuated in supine and standing positions. Their response to standing is lower than in non-overtrained athletes, with a tendency for further decreased HRV as a sign of pronounced vagal withdrawal and, in some cases, decreased sympathetic excitability, indicating a potential overtraining state. However, as a significant main characteristic, it could be noted that additional pathophysiological conditions consist in a reduced responsiveness or counter-regulation of neural drive in ANS according to an excitatory stimulus, such as an orthostatic challenge. Hence, especially active orthostatic testing could provide additional information about HR(V) reactivity and recovery giving valuable insights into athletes' training status, fatigue levels, and adaptability to workload. Measuring while standing might also counteract the issue of parasympathetic saturation as a common phenomenon especially in well-trained endurance athletes. Data interpretation should be made within intra-individual data history in trend analysis accounting for inter-individual variations in acute responses during testing due to life and physical training stressors. Therefore, additional measures (e.g., psychometrical scales) are required to provide context for HR and HRV analysis interpretation. However, incidence of orthostatic intolerance should be evaluated on an individual level and must be taken into account when considering to implement orthostatic testing in specific subpopulations. Recommendations for standardized testing procedures and interpretation guidelines are developed with the overall aim of enhancing training and recovery strategies. Despite promising study findings in the above-mentioned applied fields, further research, thorough method comparison studies, and systematic reviews are needed to assess the overall perspective of orthostatic testing for training monitoring and fine-tuning of different populations in exercise science and training.