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This study characterized by light and electron microscopy 49 photochemically induced lesions in adult rat spinal cord at 16 time intervals from 2 days to 17 months after lesioning. Vascular thrombosis, resulting from an intravascular photochemical reaction induced by a rose bengal/laser beam interaction, led within a few days to an extensive area of tissue deterioration. This area, termed the "lesion cavity" in contrast to the "secondary cavity" observed later, was at least 6 mm long and, at the epicenter, extended across most of the spinal cord width and from the dorsal surface to a level near the central canal. The area of spared tissue, 43% of the spinal cord cross-section at 2 days, did not change significantly between 2 and 56 days. Large numbers of macrophages populated the degenerating area by 5 days. This necrotic area was surrounded by a thin peripheral rim of largely intact white matter dorsally and laterally except at the epicenter where the white matter degenerated dorsomedially. In these peripheral regions, demyelination and, by 14 days, remyelination by both oligodendrocytes and Schwann cells (SCs) were evident. By 28 days, far more SCs (and meningeal cells) had entered the dorsal spinal cord, typically at the epicenter where meningeal thickening was most striking, and had migrated farther into the lesion cavity. These SCs and the axons they myelinated remained prominent in dorsal regions for many months, particularly at the epicenter; the proportion of SC to oligodendrocyte myelin diminished away from the epicenter. By 8 weeks, the lesion cavity was considerably diminished in size and thereafter it contained scattered macrophages, SC-myelinated axons, and blood vessels, primarily medially owing to flattening into clefts bilaterally. The cavity was partly bordered by astrocytes whose surfaces toward the lesion cavity were highly irregular and coated with basal lamina. Bare axons, consistently seen by electron microscopy at 5 days to 6 months, were typically ensconced among astrocytes starting at 28 days. Also by this time large, smoothly contoured, empty secondary cavities appeared, usually rostral and caudal to the epicenter; they did not increase in size or number with time. From 28 days to 17 months postlesion they occurred in 68% of the lesioned spinal cords. The secondary cavity border was composed of cells thought to be astrocytes but, surprisingly, the luminal surface was smooth and lacked basal lamina, in contrast to the primary lesion cavity border. Thus, two types of cavities formed after photochemical lesioning. This lesioning technique may provide an appropriate milieu to better understand aspects of the vexing problem of post-traumatic syringomyelia in the human.

Experimental modeling of spinal cord injury is based mostly on mechanical effects, such as the impact of a weight dropped on the exposed spinal cord. The development of the resultant lesion is influenced by many interactive factors, e.g., the efficiencies of momentum and energy transfer to the cord and their profiles in time, and consequently the histopathologic reproducibility of the lesion is often inconsistent. We describe here a recoilless method that avoids these complications (as well as laminectomy) inherently. The vascular endothelium is injured photochemically, yielding chiefly small-vessel thrombosis and associated vasogenic edema sufficient to generate spinal cord necrosis to predetermined, reproducible degrees. This model is thus intended to simulate the secondary response of the vasculature to mechanical injury In the absence of hemorrhage. The most efficient version of this technique utilizes argon-dye laser excitation of the photosensitizing dye rose bengal at its 562-nm absorption maximum in tissue. With the laser beam focused in the shape of a thin (0.3-mm) line transverse to the spinal column at T8, a narrow zone of necrosis is initially produced. Within 1 week this initial zone expands in volume (length, 6-7 mm) to create a space that, when cleared of cellular debris, is suitable for cell Implantation. In cross section, striking features are the sharp horizontal demarcation between necrosed and viable tissue and the uniform progression of lesion depth as a function of Irradiation time. The necrotic region is bordered dorsally and laterally by a thin rim of viable tissue except at the beam focus; starting at 5 days there is evidence of demyelination in this peripheral region. By 14 days, myelination by oligodendrocytes and Schwann cells begins near this rim; large numbers of Schwann cells enter the dorsal cord at the epicenter, and myelinated axons occupy previously degenerated areas. By 2 months, the initial necrotic area begins to diminish, flattening laterally into clefts. Large, empty cavities develop secondarily with luminal surfaces that are smoothly contoured, as seen by electron microscopy, in contrast to the relatively irregular border of the initial necrotic lesion. Many of these morphological attributes mirror those observed after contusion injury. We anticipate that the uniformity and reproducibility of the photochemical lesion will prove useful in conducting statistically efficient tests of strategies, such as the administration of drugs, trophic factors, or transplanted cells, which are proposed to improve outcome after spinal cord injury.

A microcomputer-based system employing photoelectric devices to record rat movements and footfalls in the grid walking test paradigm was developed and evaluated. Behavioral data obtained from the system were: distance traversed on the grid, time taken to traverse the distance, number of footfalls, times of footfalls, positions of footfalls, durations of footfalls, and whether the footfall was due to a hind or a fore limb. Validation of the system was performed by comparing the data obtained from the videotape analysis with that obtained from the computerized system. Correlation coefficients between the data obtained from the two methods were found to be 0.92 for one observer, 0.84 for a second observer, and 0.88 with the mean of the two observers. An experimental study in which a group of rats was administered dorsal hemisection lesions of the spinal cord was also conducted. Animals in the lesion group took the same amount of time to cross the runway as the control animals, but made more footfalls per crossing and had longer durations per footfall. The studies validate the capacity of the computerized system to efficiently detect fine locomotory deficits, suggesting that it is a viable tool for the evaluation of neurological dysfunctions in experimental rats.

Two inbred strains have been developed from a cross between SHR and WKY. WK-HTs are hypertensive but not hyperactive, and WK-HAs are hyperactive but normotensive. Together with SHR (that express both traits) and WKY (expressing neither trait) we used four strains to follow correlations of biological changes with the expression of hyperactivity or hypertension. We show that the well known sympathetic hyperreactivity of SHRs to acute stress is associated with the hyperactivity trait and not the hypertension among the four strains. Similarly, the well known ventricular hypertrophy in SHRs is more prominent among the hyperactive strains than the hypertensives. Examination of regional brain amine levels revealed an imbalance in forebrain serotonin transmission in the hyperactive strains, and no significant correlations with hypertension. On the other hand, neuropeptides in brainstem and spinal cord revealed a decrease, in hypertension, in neuropeptide Y and PNMT content of terminals of C1 fibers that innervate the spinal cord sympathetic outflow. Also, the two hypertensive strains showed increased TRH-and proctolin-like immunoreactivity in fibers that innervate the C1 cells in the rostral ventrolateral medulla. These findings illustrate the unique advantage provided by WK-HA and WK-HT strains as additional controls for SHRs in studying hypertension and hyperactivity.

The present study describes the production of a spinal cord lesion which is initiated by vascular occlusion resulting from the interaction between the photosensitizing dye erythrosin B and an argon laser beam. The lesion has characteristics similar to those of the central cavity thought to lead to the production of post-traumatic syringomyelia (PTS) in humans. The present study examines the behavioral and morphological characteristics of this injury over a 28-day period. Histological analysis revealed a cavity extending from the dorsal horns to lamina VIII, with some lateral and ventral pathways being spared. The cavity volume reached a maximum 7 days after lesion induction. Behavioral changes were assessed using six different tests of motor and reflex function (motor function, climbing, waterbath, inclined plane, withdrawal to pain, and withdrawal to extension). Lesioned animals exhibited flaccid paralysis for 3-5 days, which resolved afterward. The photochemically induced cavity should provide a reproducible model for examining the effects of cystic spinal cord injury on locomotor and reflex function.

Bone calcium, histomorphometry, and mechanical strength were evaluated in a model of spinal cord injury. Cortical bone area and rates of formation and apposition at the tibiofibular junction (TFJ) and midshaft of the humerus were measured at 35-42, 42-77, and 77-94 days after transection of the spinal cord. All comparisons were between the animals with a spinal lesion and control animals. A 0.34% difference in the length of the tibia of the two groups of animals was observed, the dry weight of the tibia was 28.4% less in rats with a lesion, and there was no significant difference in the amount of calcium per milligram of bone. At 35 days after surgery, the cortical area in the midshaft of the humerus was slightly less (11%) in rats with a lesion, but by 94 days there was no difference in cortical or medullary area. The final (day 94) cortical area at the TFJ in rats with a lesion was less than that in the controls. Bone formation at the TFJ was similar in both groups in the period of 35-42 days and was similar at 77-94 days in the animals with a lesion, and rates of formation and apposition were greater in the humerus of rats with a lesion; the rates did not differ significantly between groups at either site. At 94 days, trabecular bone area in the tibial metaphysis was 41% less in rats with a lesion. Mechanical parameters were significantly less in the femora, but not the humeri, of rats with a lesion.

A double-peroxidase procedure was used to study the ultrastructural relationships between terminals and fibers containing three putative neurotransmitters and retrogradely identified sympathetic preganglionic neurons (SPNs) located in the intermediolateral cell column (IML) of the rat. SPNs with axons in the cervical sympathetic trunk were retrogradely labeled with horseradish peroxidase. In addition, terminals and fibers containing substance P, enkephalin, and serotonin were detected using immunohistochemistry. Sections containing both retrogradely labeled SPNs and immunoreactive processes were processed for electron microscopy. Ultrastructural examination revealed synaptic contacts between terminals containing each of these three neurotransmitters and retrogradely labeled dendrites from SPNs. Also, immunoreactive terminals were apposed to retrogradely labeled cell bodies. Therefore, these transmitters may alter sympathetic function by their direct action on SPNs.

post-lesion. The present study describes the competition and interaction between sprouting primary afferent fibers and descending catecholaminergic fibers with the outgrowth from an intraspinal transplant of fetal raphe neurons transplanted either 7 days or 3 months post-lesion. Adult Sprague-Dawley rats received an intrathecal injection of 5,7-dihydroxytryptamine (5,7-DHT) at the C6 spinal cord level to lesion the descending serotonergic system. At 7 days or 3 months post-lesion, cell suspensions of E14.515 fetal raphe cells were transplanted at the T8 spinal cord level. The control groups included unoperated controls, sham 5,7-DHT lesioned animals, and 5,7-DHT lesioned animals that. received sham transplants at either 7 days or 3 months post-lesion. The immunohistochemical localization of serotonin (5-HT), calcitonin gene-related peptide (CGRP), dopamine /3-hydroxylase (D/3H) and phenylethanolamine N-methyltransferase (PNMT) was performed at 1, 3, 6, 12 and 18 months post-transplant. A Zeiss IBAS-2000 image analysis system was used to quantify the percent area of the IML that was covered by 5HT, CGRP-, DflH, and PNMT-immunofluorescent fibers. The total cross sectional areas of the spinal cords taken from all the groups indicated that there was no change in the total area of the spinal cord or of the. IML region following the 5,7-DHT lesion and the transplant. Animals with 5,7-DHT lesions showed exten

A proctolin (PROC)-like peptide was studied immunohistochemically in the hypothalamus, lower brainstem and spinal cord of the rat using an antiserum against PROC conjugated to thyroglobulin. Neuronal cell bodies containing PROC-like immunoreactivity (PROC-LI) were observed in the dorsomedial, paraventricular and supraoptic nuclei of the hypothalamus and in the nucleus raphe magnus, nucleus raphe pallidus, nucleus raphe obscurus and nucleus interfascicularis nervi hypoglossi in the medulla oblongata. Fibers containing PROC-LI were seen in the median eminence and in other hypothalamic nuclei, and in the lower brainstem in cranial motor nuclei including the dorsal motor nucleus of the vagus nerve, the motor trigeminal nucleus, the facial nucleus and nucleus ambiguous, and in lower numbers in the nucleus of the solitary tract and locus coeruleus. Fibers containing PROC-LI were also located in the spinal cord, in the intermediolateral cell column at thoracic levels and in the ventral horns at all levels of the spinal cord. After transection of the spinal cord, all PROC-immunoreactive fibers below the lesion disappeared. Following injection of Fast blue into the thoracic spinal cord, retrogradely labeled cells in the nuclei raphe pallidus, obscurus and magnus and nucleus interfasciculari nervi hypoglossi were seen to contain PROC-LI. PROC-LI had a similar distribution as thyrotropin-releasing hormone (TRH)-LI in the above-mentioned areas and coexistence of TRH-LI and PROC-LI was shown in cell bodies in the hypothalamus and medulla oblongata. PROC-LI could also be shown to coexist with 5-hydroxytryptamine (5-HT)-LI in neuronal cell bodies in the lower brainstem. The results demonstrate the occurrence of a PROC-like peptide in the mammalian nervous system, and these neurons seem to be at least largely identical to previously described TRH systems. A possible involvement of the PROC-like peptide in spinal motor control is discussed in relation to the well-established role of PROC in control of motor behavior in insects and invertebrates.