Objective This study was undertaken to characterize structural and pharmacological properties of the pig popliteal artery in order to develop a novel system for the examination of lower limb blood flow regulation in a variety of cardiovascular pathologies, such as diabetes‐induced peripheral artery disease. Methods Popliteal arteries were isolated from streptozocin‐induced diabetic pigs or age‐matched saline‐injected control pigs for morphological study using transmission electron microscopy and for examination of vasoreactivity to pharmacological agents using wire myography. Results Transmission electron microscopy of the porcine popliteal artery wall revealed the presence of endothelial cell‐smooth muscle cell interactions (myoendothelial junctions) and smooth muscle cell‐smooth muscle cell interactions, for which we have coined the term “myo‐myo junctions.” These myo‐myo junctions were shown to feature plaques indicative of connexin expression. Further, the pig popliteal artery was highly responsive to a variety of vasoconstrictors including norepinephrine, phenylephrine, and U46619, and vasodilators including acetylcholine, adenosine 5′‐β‐thio diphosphate, and bradykinin. Finally, 2 weeks after streptozocin‐induced diabetes, the normalized vasoconstriction of the pig popliteal artery to norepinephrine was unaltered compared to control. Conclusions The pig popliteal artery displays structural and pharmacological properties that might prove useful in future studies of diabetes‐associated peripheral artery disease and other lower limb cardiovascular diseases.