Extraocular muscles of the rat possess numerous nerves suitable for the study of fine structure. In these muscles, small nerves made up of one to ten myelinated and unmyelinated nerve fibers are surrounded by two or three layers of perineurium. The perineurium is arranged in concentric sleeves, each one cell thick. Continuous boundary membranes separate the perineural sleeves from the epineural and endoneural tissue space, but the boundary membranes may be spotty or absent between individual sleeves. The presence of boundary membranes around perineural cells distinguishes them from nearby fibroblasts which lack similar membranous investment. Tight intercellular junctions join the cells comprising each sleeve so that the nerves are completely ensheathed in perineurium. The number of sleeves decreases as the nerve becomes smaller, either by the termination of the innermost sleeve or by the loss of a sleeve as the nerve branches. The last sleeve ends shortly before the termination of the nerve. The perineurium is thus open-ended peripherally and, at these places, the epineurium and endoneurium are continuous. Continuities between the epineurium and endoneurium also exist at the entrance and exit of blood vessels supplying the nerve and at points where reticular fibers pierce the perineurium. These structural features correlate well with the action of the perineurium as a diffusion barrier and as a pathway in the transmission of infections.
Use of the capillary bed of skeletal muscle as an in vivo recipient site to transplant autologous endothelial cells that have undergone gene transfer ex vivo has considerable potential as a technique of somatic gene therapy. Here we document a previously unrecognized capacity of endothelial cells to adhere and incorporate spontaneously into confluent endothelial cell monolayers in vitro and in vivo. This spontaneous adhesion and incorporation of endothelial cells enabled us to seed lacZ-transduced endothelial cells into the wall of skeletal muscle capillaries of the hindlimb of the rat. Certain transduced endothelial cells became incorporated within the capillary wall, whereas others remained within the capillary lumen where they formed focal, electron-dense, contacts with host endothelium. lacZ expression in the capillary bed was documented for up to 1 month after transplantation. Use of the intact capillary bed of skeletal muscle as an in vivo recipient site for transduced, autologous endothelial cells holds promise as a strategy for somatic gene therapy to treat various genetic and acquired human diseases.
The endothelium plays an important role in mediating vasodilator effects of several agents (acetylcholine, thrombin, A23187, etc.). The goal of this study was to determine the ability of oxygen free radicals generated by electrical field stimulation to alter endothelial function in isolated tissue systems. Tail artery strips and the mesenteric microvasculature isolated from Sprague-Dawley rats were used. Following smooth muscle contraction induced by norepinephrine, these preparations relaxed in response to acetylcholine chloride or ionophore A23187. All vessels were then subjected to electrical stimulation (9 V, 1-2 ms, 4 Hz) of the physiological buffer in which they were bathed or perfused. In some of these preparations, an antioxidant, (10(-4) M sodium ascorbate, 3.6 X 10(-5) M glutathione, 1.3 X 10(-2) M dimethyl sulfoxide) was included in the buffer. Relaxation responses persisted in vessels where an antioxidant had been included in the electrically stimulated buffer. Tissues stimulated without this protection did not relax on subsequent exposures to endothelium-dependent vasodilators. Scanning-electron microscopy of the tissues revealed significant endothelial damage (cell membrane pitting) in tissues exposed to electrical stimulation without antioxidant protection. These results suggest that electrical stimulation causes endothelial damage in isolated vascular preparations. This seemingly adverse effect proves to be a useful tool for removing the endothelium in studies of isolated vascular tissues.
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