Introduction: Pharmacological evaluation of the unique equine laminar microvasculature is crucial to understanding its role in health and in diseases such as laminitis. However, separating the distinctive characteristics of arterial versus venous components of this complex vascular network has previously proved to be extremely difficult. Encased in a hard hoof capsule, isolation of individual blood vessels presents a considerable challenge. Exacerbating this difficulty, the laminar venous network is adapted to sustain high intravascular pressures and consequently has thickened walls, making the normally straightforward visual distinction between arteries and veins problematic. Here we describe a novel harvesting and dissection method coupled with a functional analysis procedure that facilitates distinction of arteries and veins.
Methods: Laminar tissue was recovered from the hoof of euthanized, clinically normal horses by dissection at the coronary band and stored in cold Krebs-Henseleit physiological salt solution prior to further dissection in the laboratory to remove 2 mm segments of vessels 100-500 microm in diameter. Active length tension measurements were made to evaluate optimal conditions for experimentation, and based on the differences in contractility and appearance, an experimental protocol was set up to allow a) initial distinction between arteries and veins and b) in vitro pharmacological evaluation.
Results: Active length tension studies clearly revealed the presence of two populations of vessels distinguished by either a large or a lower maximal contraction that subsequent histological evaluation confirmed to be arteries and veins respectively. Functional distinction using relative contractility to 60 mM potassium salt solution then demonstrated equine laminar veins to have increased sensitivity to the agonist endothelin 1 (ET-1) compared to arteries.
Discussion: In vitro evaluation of laminar vessels is possible despite anatomical obstacles. Furthermore, a clear distinction can be made between laminar veins and arteries using functional characteristics providing vessels of a similar size range are selected. Utilising these novel procedures, investigators can unambiguously analyse the pharmacological characteristics of equine laminar veins and arteries to decipher the physiological mechanisms responsible for the control of laminar blood flow.