Michael P. Kaye

Patterns of Sympathetic Nerve Projections onto the Canine Heart

Major branches from right and left sympathetic ganglia were electrically stimulated while force of contraction was recorded from multiple areas of the right and left ventricles. Stimulation of the stellate ganglia generally elicited alterations in force of contraction from all test segments, but excitation of selected nerve trunks induced responses in highly localized regions of the heart. Ablation of narrow strips of epicardium resulted in obliteration of contractile responses in specific, highly localized regions of the heart; thus a major fraction of the sympathetic innervation of the ventricular chambers is by way of the epicardial plexus. The anterior surface of the right ventricle is supplied by projection pathways arising within the immediately subepicardial regions of the right A-V groove and, to a lesser extent, from the tissues immediately adjacent to the left anterior descending artery. The left ventricle receives minor projections from the right A-V groove with major projections from subepicardial tissues along the left anterior descending artery. In some animals there also exists a definite left ventricular supply from the region of the left A-V groove. Whereas the thoracic vagi send dense projections to the atria, and particularly to nodal tissue, they also supply both ventricles with inhibitory and augmentor fibers. Although isolated cardiac nerves may carry predominantly sympathetic or parasympathetic fibers, many show rich intermingling of these fibers in trunks distal to the caudal cervical ganglion.

Cardiac dysrhythmias in the conscious dog after surgically induced autonomic imbalance

The ventrolateral cardiac nerve in the dog is a primary branch of the left sympathetics and represents a direct neural link between the central nervous system and the heart. Its electric excitation elicits characteristic shifts in pacemaker and tachydysrhythmias related to its explicit innervation of the inferior atrial, atrioventricular (A-V) junctional and ventricular tissues. Total denervation of the canine heart, sparing the ventrolateral cardiac nerve, produced a long-term model in which only these portions of the heart retained their sympathetic innervation. The trained unanesthetized model dog was subjected to severe exercise in order to determine the effects of elevated levels of sympathetic tone upon these important regions of the conduction system. Reproducible tachydysrhythmias were elicited in all six animals completing the regimen of periodic testing over a period of 136 to 378 days after operation. The abnormal rhythms consisted of shifting cardiac pacemakers and supraventricular A-V junctional and ventricular tachycardias with frequent premature systoles. Comparable abnormalities were not observed in a similarly tested sham-operated animal or in dogs with a totally denervated heart. The exercise-induced dysrhythmias gradually disappeared with time, presumably in relation to autonomic reinnervation of the heart. The characteristic patterns of ventrolateral cardiac nerve and upon its presumed influence upon Purkinje fiber and A-V nodal automaticity and temporal dispersion of refractoriness in myocardial tissues.

Autonomic Neural Control of Cardiac Rhythm: The Role of Autonomic Imbalance in the Genesis of Cardiac Dysrhythmia

Cardiac dysrhythmias result from abnormalities in rate, regularity, or sequence of cardiac activation, and because of direct actions of the autonomic nervous system upon each of these properties, imbalance in this system may play an important role in the genesis of cardiac dysrhythmia. A canine model has been developed in which the extrinsic innervation of the heart is ablated with the exception of the ventrolateral cardiac nerve. This nerve is distributed primarily to the inferior atrial, AV junctional, and ventricular tissues. Following recovery from surgery, the animal is placed on a treadmill and required to perform strenuous exercise. In all of six animals which sustained repeated exercise testing over periods of 4-12 months, dysrhythmias of varying complexities were elicited. None appeared in parallel experiments conducted in control or sham-operated animals. The dysrhythmias consisted of supraventricular, AV junctional, or ventricular tachycardias with occasional premature atrial or ventricular systoles. The dysrhythmias were not influenced by atropine but were generally controlled by propranolol.

Radioisotopic techniques for noninvasive detection of platelet deposition in bovine-tissue mitral-valve prostheses and in vitro quantification of visceral microembolism in dogs.

Platelet deposition on bovine pericardial-tissue mitral-valve prostheses in 11 dogs was observed noninvasively by use of 111In-labeled platelets and quantified after sacrifice at one (n = 3), 14 (n = 3), and 30 (n = 5) days postimplantation (300-400 microCi of labeled platelets having been injected 24 hours previously). Thrombosis on the sewing ring and pericardial leaflets at one and 14 days and on the leaflets at 30 days was delineated in scintiphotos. In vitro quantification (% injected dose) indicated that the leaflets, sewing ring, and perivalvular tissue retained 0.904% of labeled platelets at one day postimplantation, 0.198% at 14 days, and 0.040% at 30 days. Platelet half-life was reduced to 38 hours at 21 days postimplantation but returned toward the normal (50 hours) with fibrous ingrowth in the sewing ring. Microembolism in lung and kidney, as measured by tissue/blood radioactivity ratio, also was decreased significantly at 30 days. 111In-labeled platelets thus provide a sensitive marker for noninvasive imaging and in vitro quantification of platelet deposition on valvular prostheses and microemboli trapped in viscera, although histochemical confirmation will be necessary to correlate the increase in tissue/blood ratio with the presence of microembolism.

Prolonged Pressor Effects of Selective Stimulation of the Stellate Ganglion

With both intensity and duration of individual current pulses maintained constant and at low values, frequencies were varied from ½ to 80/sec., and the stimulating current was applied to the stellate ganglion of the open chest dog. Whereas systolic and pulse pressures increased rapidly with the application of high frequency currents, the pressures were not long maintained even though the stimulation was continuous. With low frequency currents, systolic and pulse pressures rose more slowly but to higher levels which were sustained for periods of 5 to 11 hours. It is believed that neither cardiac acceleration nor systemic vasoconstriction contributed significantly to the pressor responses, but rather that augmentation of myocardial contraction and increased systolic ejection were the important causative factors.

The Unbalanced Heart

The heart often appears to function independently because it can maintain regular activity even when isolated from the rest of the body. Likewise, it maintains its regularity in the midst of powerful

Quantification of Deposition of Neutrophilic Granulocytes on Vascular Grafts in Dogs With 111In-Labeled Granulocytes

A new radioisotopic technique has been developed for quantification of deposition of neutrophilic granulocytes on vascular grafts. Nine healthy mongrel dogs underwent bilateral femoral artery resection and reconstruction with grafts of femoral vein and Gore-Tex. Pure granulocytes that had been separated from whole blood by centrifugal elutriation were labeled with 111In-tropolone in plasma. The granulocyte harvesting efficiency was 25 +/- 12%, and the labeling efficiency was 87 +/- 7%. Three hours after injection of labeled granulocytes and 2 hours after reperfusion, the grafts were harvested and cut into several segments for study of areas of anastomoses and midsections. On the basis of the radioactivity in the blood and in anastomotic and graft sections, the area of graft sections, and the neutrophilic granulocyte and differential leukocyte counts, the number of neutrophilic granulocytes adherent to a unit area and the total number of neutrophilic granulocytes on graft sections were calculated. These quantifications of the deposition of neutrophilic granulocytes indicated that the midsections of Gore-Tex grafts retained more neutrophilic granulocytes than did the midsections of vein grafts. Although the anastomotic areas retained more neutrophilic granulocytes than did the midsections of vein grafts, the opposite finding prevailed for the Gore-Tex grafts. A major fraction of neutrophilic granulocytes on Gore-Tex grafts was incorporated into thrombus. Semiquantitative information obtained by scintigraphy of the deposition of neutrophilic granulocytes on vascular grafts also confirmed this observation.

Cardiac Effects of Nerve Growth Factor in Dogs

Nerve growth factor, a protein necessary for the growth and maintenance of sympathetic nerves, was administered to newborn puppies and to pregnant bitches. Chemical analysis of the hearts of these animals at 3 months of age revealed a marked increase in myocardial norepinephrine. Gross and microscopic studies suggested myocardial hypertrophy. These changes occurring in the absence of systemic hypertension and caused by administration of nerve growth factor to dogs in the prenatal period raise questions concerning a potential role of nerve growth factor in the spontaneous development of myocardial abnormalities.

Late Thrombogenicity of Grafts in the Venous System

Although some previous experiments have demonstrated encouraging early results with venous vascular prosthesis, the long-term results have not been sufficiently investigated. The late thrombogenicity of 12 autogenous vein grafts and eight polytetrafluoroethylene (PTFE) grafts, three of which had been subjected to endothelial seeding, was evaluated 6 months after implantation in the canine venous system by determination of the survival time of the indium-labelled autogenous platelets and of the light and electron microscopic appearance of the graft-to-blood interface. Twelve dogs receiving autogenous vein grafts had a platelet half-life of 64 ± 13 (mean ± s.d.) hours and a completely thrombus-free internal graft surface covered by a smooth endothelium. The platelet half-life in five dogs with non-seeded PTFE grafts was significantly (P < 0.01) shorter (44 ± 11h). The PTFE-to-blood interface consisted of a 100- to 170-μm-thick pseudointima with a thrombus-free surface area of 25-90% and substantial endothelial coverage in only two animals. Three dogs receiving endothelial-seeded PTFE grafts had a platelet half-life of 38, 46 and 53h, respectively, a 10- to 80-μm-thick pseudointima covered with typical endothelium, and a thrombus-free surface area of 90-100%. Our results demonstrate that PTFE grafts are still thrombogenic at 6 months following implantation in the canine venous system. Therefore, additional measures are required to maintain graft patency. Our limited experience with endothelial seeding does not allow any definite conclusions, but our findings indicate that this procedure might be beneficial.