Obesity has been a major public health problem worldwide and it has reached epidemic levels in the past few decades. It is also a significant contributor to cardiovascular disease, development of hypertension, and microvascular target organ damage. Although the underlying mechanism is unclear, endothelial dysfunction has been attributed to obesity which, in turn, can disrupt the vasodilation of the small arteries. The Endothelial Dependent Hyperpolarization (EDH) is the predominant vasodilatory mechanism in the small resistance arteries. EDH is tightly controlled by brief local Calcium (Ca2+) signals: Ca2+pulsars, which are IP3-mediated, and Ca2+ sparklets that reflect Ca2+ entry through TRPV4 channels. Studying Ca2+ signals is critical in obesity- related research due to its direct impact on EDH and on endothelial health. However, no studies have been conducted to address endothelial local Ca2+ signals as related to obesity in humans. Therefore, this research is aimed at measuring the changes to endothelial Ca2+ signaling (Ca2+ pulsars and Ca2+ sparklets) in human obesity and relate this to the vasodilatory capacity of the vascular endothelium. A rigorous approach and study protocol were developed to address the use of human arteries that were isolated from omental biopsies taken from healthy women undergoing elective cesarean sections at terms (37-42 weeks gestation). Isolated omental arteries were examined using high speed spinning disk confocal microscopy (Andor XD Revolution) for imaging endothelial Ca2+ events in an en-face configuration. Endothelial functional studies were carried out using four-channel wire myograph system (Danish MyoTech) to assess wire-induced stretch and agonist effects. Results have shown that in human endothelium, Bradykinin (30 nM) induced an increase in pulsars frequency (n=6). Pulsars frequency was unaffected by obesity (n=5). In regard to Ca2+ sparklets, imaging results have shown that TRPV4 Ca2+ sparklets could not be imaged by directly activating TRPV4 channels using GSK101 (100 nM) in the human endothelium. However, the myography studies have demonstrated that TRPV4 activator GSK101 (1-100 nM) dose dependently relax pre-constricted omental artery segments (n= 5). GSK101- induced relaxation was abolished by the large conductance potassium channels inhibitor Paxilline (10 μM) (n=6) in the Vascular Smooth Muscle Cell (VSMC). GSK101 is believed to have an agonistic effect on the TRPV4 channels in the VSMC and causes vasodilation of the artery. These findings serve as a fundamental basis for better understanding the mechanism of EDH in human arteries and represent a promising area for future research.