Structural recovery in lesioned adult mammalian spinal cord by x-irradiation of the lesion site.

Mechanical injury to the adult mammalian spinal cord results in permanent morphological disintegration including severance/laceration of brain-cord axons at the lesion site. We report here that some of the structural consequences of injury can be averted by altering the cellular components of the lesion site with x-irradiation. We observed that localized irradiation of the unilaterally transected adult rat spinal cord when delivered during a defined time-window (third week) postinjury prevented cavitation, enabled establishment of structural integrity, and resulted in regrowth of severed corticospinal axons through the lesion site and into the distal stump. In addition, we examined the natural course of degeneration and cavitation at the site of lesion with time after injury, noting that through the third week postinjury recovery processes are in progress and only at the fourth week do the destructive processes take over. Our data suggest that the adult mammalian spinal cord has innate mechanisms required for recovery from injury and that timed intervention in certain cellular events by x-irradiation prevents the onset of degeneration and thus enables structural regenerative processes to proceed unhindered. We postulate that a radiation-sensitive subgroup of cells triggers the delayed degenerative processes. The identity of these intrusive cells and the mechanisms for triggering tissue degeneration are still unknown.

Radiation-induced reductions in macrophage recruitment have only slight effects on myelin degeneration in sectioned peripheral nerves of mice.

Macrophage recruitment into the distal nerve stump of the cut or crushed sciatic or saphenous nerves of C57BL/6J mice was reduced by prior whole body irradiation. This procedure was successful in keeping the numbers of cells stained with the mouse macrophage-specific antibody F4/80 to the levels found in unsectioned nerves. Quantitative image analysis of immunostained sections showed that the rate of loss of myelin basic protein was identical in nerves from irradiated and unirradiated mice up to 5 days but thereafter was slower in macrophage-deprived nerves. Similar analysis of semithin sections stained with toluidine blue detected more undegenerated myelin in the nerves from irradiated mice 10 days after operation. Quantitative counts made from electron micrographs of the sectioned nerves at 7 days also showed slightly less extensive myelin breakdown in the nerves from irradiated mice. Complete removal of myelin from some Schwann cells can occur without macrophages, but macrophages accelerate the removal of myelin in the later stages of Wallerian degeneration. It is concluded that there are two phases to the breakdown of myelin in peripheral nerves undergoing Wallerian degeneration: an initial stage entirely dependent on the activity of Schwann cells and a later stage dependent on both Schwann cells and the presence of macrophages.

Dye-induced photolesion in the mammalian retina: glial and neuronal reactions.

Irradiation in the presence of a dye applied to the extracellular space is known to produce degenerative features in irradiated neurones and fibers. In the present study, we confirmed the potential use of this procedure as a lesion technique by showing the removal of degenerating elements as part of the glial reaction to the lesion. The dye Rose Bengal was applied to the vitreous body of a rat eye and a T-shaped irradiation pattern was projected onto the retina within the absorption band of the dye. Degenerative features were restricted to the irradiated area, which could be readily identified from its shape. Retinae examined after various survival times showed that macrophages invaded the damaged area within 1 day, and that mitotic activity of reactive glial cells subsequently occurred in the vicinity of the wound. Both cell types were identified by their structural features. Macrophages were also revealed by a staining technique using the dye Nile Red, whereas reactive glial cells were immunolabeled with an antibody directed against the glial fibrillary acidic protein. Reactive glial cells helped the macrophages to gradually remove injured cells and damaged processes. Their main task, however, appeared to be in scar formation, since their processes seemed to suture the lips of the wound together and restore the limiting membrane at the inner retina. After 2 months' survival time, the parent ganglion cells of most disrupted axon bundles had retrogradely degenerated, but regenerating ganglion cell axons were also observed. These results provide some new data about healing processes in the retina. They demonstrate that the dye-induced photolesion technique can be used to either remove or axotomize selected neurones in neural networks which have been made optically accessible.

[Experimental study on intraoperative irradiation for pancreatic cancer: histological and nutritional changes in early phase after high dose irradiation].

For the treatment of pancreatic head cancer, pancreatoduodenectomy is followed by the intraoperative radiation therapy (IORT). The present dose adopted ranged 20 to 30 Gy, however the dose is believed to be insufficient for local control of adenocarcinoma cells. In this study, high dose IORT was performed using rabbit, so histological and nutritional changes were evaluated. Rabbits were divided in three groups; 30 Gy, 50 Gy, 80 Gy. The radiation was performed with electrons focusing base of cranial mesenteric artery (SMA in human). The rabbits were sacrificed at intervals ranging from immediately after to 4 weeks following irradiation. The earliest evidence of histological changes was the loss of endothel, although it was repaired within 1 week. Fragmentation and reduplication of internal elastic lamina were observed after 1 week, however the degree was not dose dependent. Damages of the media was observed in 50 and 80 Gy groups. That is, focal degeneration of smooth muscle cell was demonstrated in 50 Gy group and medial necrosis in 80 Gy group. Degeneration of ganglion cells was observed and its severity was dose dependent. In 80 Gy group, diarrhea occurred more frequently compared with the other groups and body weight loss couldn't recover within 4 weeks. It is concluded that, since necrosis of aortic media and marked degeneration of ganglion cells are inevitable in 80 Gy group, IORT dose should be increased within 50 Gy.

Dose and temporal parameters in delaying injured optic nerve degeneration by low-energy laser irradiation.

Low-energy laser irradiation has been reported to postpone the degenerative processes in crushed optic nerves of rats, which are part of the nonregenerable mammalian central nervous system. In the present study, we evaluated the optimal irradiation parameters for this purpose. Optic nerves of 141 rats were subjected to crush injury and then irradiated through the eye, starting at different points of time before or after the injury, for different durations and periods, using various intensities of either helium-neon laser or noncoherent infrared light (904 nm). The effect was evaluated by measurements of the compound action potentials of the nerve segments between the site of injury and the optic chiasm. The compound action potential amplitude of the crushed nonirradiated nerves, as measured 2 weeks after the injury, was found to be 0.51 +/- 0.30 mV, in contrast to 3.10 +/- 1.03 mV measured in 232 normal nerves. Irradiation with a 10.5 mW helium-neon laser for 2 and 3 min once a day for 14 consecutive days resulted in maximal preservation of action potentials (1.78 +/- 0.72 and 1.95 +/- 0.71 mV, respectively). Irradiations beginning immediately prior to the injury were as effective as irradiations beginning soon after it. Irradiations for longer than 3 min or twice a day aggravated the damage. Noncoherent infrared light was ineffective or adversely affected the injured nerves. Our experiments suggest that optimal delay of posttraumatic optic nerve degeneration in rats is attainable with 10.5 mW helium-neon laser irradiations for 2 or 3 min once a day for 14 consecutive days.

Memory-based learning in preweanling and adult rats after infantile x-irradiation-induced hippocampal granule cell hypoplasia.

Infantile exposure to x-irradiation induced severe hippocampal granule cell hypoplasia in preweanling and young adult rats. Hippocampally damaged pups, tested at 16 days of age, showed deficits in a memory-based discrimination based on single alternations of reward and nonreward when training was conducted at a 60-s intertrial interval (ITI) but not when conducted at a 30-s ITI. This deficit was still present at the 60-s ITI in animals x-irradiated in infancy and tested at 60-65 days of age. These data provide further support for the role of the hippocampus in intermediate-term memory and demonstrate, in a developmental context, the importance of an intact hippocampus in learning that depends on nonspatial memory.

Beneficial effects of x-irradiation on recovery of lesioned mammalian central nervous tissue.

We examined the potential of x-irradiation, at clinical dose levels, to manipulate the cellular constituents and thereby change the consequences of transection injury to adult mammalian central nervous tissue (rat olfactory bulb). Irradiation resulted in reduction or elimination of reactive astrocytes at the site of incision provided that it was delivered within a defined time window postinjury. Under conditions optimal for the elimination of gliosis (15-18 days postinjury), irradiation of severed olfactory bulbs averted some of the degenerative consequences of lesion. We observed that irradiation was accompanied by prevention of tissue degeneration around the site of lesion, structural healing with maintenance of the typical cell lamination, and rescue of some axotomized mitral cells (principal bulb neurons). Thus radiation resulted in partial preservation of normal tissue morphology. It is postulated that intrusive cell populations are generated in response to injury and reactive astrocytes are one such group. Our results suggest that selective elimination of these cells by irradiation enabled some of the regenerative processes that are necessary for full recovery to maintain their courses. The cellular targets of these cells, their modes of intervention in recovery, and the potential role of irradiation as a therapeutic modality for injured central nervous system are discussed.

Temporal parameters of low energy laser irradiation for optimal delay of post-traumatic degeneration of rat optic nerve.

Compression injury of a central nerve results in its degeneration with irreversible loss of function due to the inability of the mammalian central nervous system (CNS) to regenerate. In contrast, the CNS of lower vertebrates has a high capacity to regenerate. Recently, low energy laser irradiation was shown to attenuate degeneration in injured CNS nerves. The optic nerves of rats were subjected to moderate crush, calibrated so that some electrophysiological activity was preserved. The nerves were then subjected to low energy laser irradiation (10.5 mW, 2 min daily) for various periods. The electrical activity of the nerves, distal to the site of injury, was determined by measuring the compound action potential at the termination of the experiment. Two weeks of irradiation begun immediately after injury and continued daily thereafter, resulted in a compound action potential which was significantly higher (mean +/- S.E.M. 1856 +/- 535 microV) than that of non-irradiated injured nerves (351 +/- 120 microV). The effect was temporary and subsided within a week. This two-week irradiation was slightly more effective than a treatment lasting one week (1406 +/- 225 microV) and was significantly more effective than 4 days of irradiation (960 +/- 133 microV). The number of treatments is therefore important. The time at which the treatment commences relative to the injury is also critical. Irradiation initiated two hours after the crush was about half as effective as immediate irradiation (810 +/- 42 microV). No apparent effect was evident when the laser was applied for the first time 5 h, or longer, after the crush.(ABSTRACT TRUNCATED AT 250 WORDS)

An ultrastructural study of the geniculo-suprachiasmatic projection in the rat

A projeþõo geniculo-supraquiasmática foi estudada em ratos, examinando-se, ao microscópico eletr¶nico, as terminaþ÷es sinápticas em degeneraþõo que aparecem no núcleo suprachciasmático depois que uma lesõo por radiofrequÛncia é feita no núcleo geniculado lateral ventral. Os elementos pré-sinápticos em degeneraþõo eram terminais de ax¶nios e os elementos pós-sinápticos eram geralmente dendritos. Vesículas sinápticas claras, pleomórifcas e, as vÛzes, vesículas granulares eram visíveis nas terminaþ÷es axonais em degeneraþõo. As sinapses degeneradas eram simétricas, ou do tipo II de Gray (AU)

An ultrastructural study of the geniculo-suprachiasmatic projection in the rat

A projeçäo geniculo-supraquiasmática foi estudada em ratos, examinando-se, ao microscópico eletrônico, as terminaçöes sinápticas em degeneraçäo que aparecem no núcleo suprachciasmático depois que uma lesäo por radiofrequência é feita no núcleo geniculado lateral ventral. Os elementos pré-sinápticos em degeneraçäo eram terminais de axônios e os elementos pós-sinápticos eram geralmente dendritos. Vesículas sinápticas claras, pleomórifcas e, as vêzes, vesículas granulares eram visíveis nas terminaçöes axonais em degeneraçäo. As sinapses degeneradas eram simétricas, ou do tipo II de Gray

[Changes in synapses after gamma-irradiation of the rat head]. / Izmeneniia sinapsov pri gamma-obluchenii golovy krys.

High reactivity and, at the same time, flexibility of interneuronal contacts were observed after exposure of rat head to 2-100 Gy radiation. At high doses (200-400 Gy) radiation-induced changes played a major role in the development of the cerebral form of radiation sickness. A complete asynapsis is probably one of the causes of the animals death "under the ray" (irradiation of the head with a dose of 1000 Gy).

Stimulatory effect of He-Ne low dose laser on injured sciatic nerves of rats.

Injury to a mammalian peripheral nerve is accompanied by a restorative process that is manifested after a delay. This process is expressed morphologically by the emergence of new nerve fibers. Restoration of function occurs when the regenerating fibers reconnect with the target organ. Because of the low rate of fiber elongation, the denervated target is partially degenerated by the time that the regenerating fibers approach it. To prevent such an atrophy, one must find a way to prevent the degeneration of the nerve, to speed up regeneration, or to maintain the target during the period of nerve degeneration. In the present work, we examined the potential of treatment with low energy laser radiation for improving regeneration or preventing degeneration of mammalian peripheral nerve after injury. After repeated injury for 20 consecutive days, treatment of the sciatic nerve of the rat with low energy laser (He-Ne, 17 mW) caused a significant increase in the amplitude of the action potential recorded in the corresponding gastrocnemius relative to the action potential of injured but not treated nerves. The action potential of the injured sciatic nerves that were laser-irradiated increased to values close to that of a noninjured nerve. The studies include follow-up for 1 year after the injury. This electrophysiological manifestation of the effect of laser treatment on injured nerves was accompanied by a diminution of the size of the scar tissue from these nerves. Yet to be resolved is whether these two phenomena (i.e., electrophysiological and morphological responses) coincide or whether they relate to each other.(ABSTRACT TRUNCATED AT 250 WORDS)

Laser influence on the rate of degeneration of damaged nerves.

The sciatic nerves were sectioned and sutured back. The rate of degeneration at the point of suture was studied. Laser treatment increased the rate of degeneration of the damaged fibers.

Effects of low-energy He-Ne laser irradiation on posttraumatic degeneration of adult rabbit optic nerve.

Axons of the mammalian peripheral and central nervous systems degenerate after nerve injury. We have recently found that He-Ne laser irradiation may prevent some of the consequences of the injury in peripheral nerves of mammals. In the present study, the efficacy of the laser in treating injured neurons of the mammalian CNS was tested. Optic nerves of adult rabbits were exposed daily for 8-14 days to He-Ne laser irradiation (14 min, 15 mW) through the overlying muscles and skin. As a result of this treatment, the injured nerves maintained their histological integrity, which is invariably lost in injured mammalian CNS neurons.

Delayed effects of ionizing radiation on the ear.

The question of damage to the ear from exposure to ionizing radiation was addressed by exposing groups of chinchillas to fractioned doses of radiation (2 Gy per day) for total doses ranging from 40 to 90 Gy. In order to allow any delayed effects of radiation to become manifest, the animals were sacrificed two years after completion of treatment and their temporal bones were prepared for microscopic examination. The most pronounced effect of treatment was degeneration of sensory and supporting cells and loss of eighth nerve fibers in the organ of Corti. Damage increased with increasing dose of radiation. The degree of damage found in many of these ears was of sufficient magnitude to produce a permanent sensorineural hearing loss.

A sensitive assay for detecting hypersensitivity to ionizing radiation in lymphoblastoid lines from patients with Duchenne muscular dystrophy and primary neuronal degenerations.

Hypersensitivity to the lethal effects of DNA-damaging agents is usually demonstrated using the classical colony-forming ability assay with cultured fibroblast lines. Based on the ability of viable cells in lymphoblastoid lines (Epstein-Barr virus-transformed B lymphocytes) to exclude the vital dye trypan blue, we have developed a more rapid survival assay which has been useful in detecting hypersensitivity to ionizing radiation in certain diseases characterized by primary degeneration of excitable tissue. We now present a complete description of this post-X-ray survival assay. We also demonstrate the suitability of both our assay and our method of data analysis for detecting hypersensitivity to ionizing radiation. This demonstration is based on a detailed analysis of assay results with lymphoblastoid lines from 28 normal donors, 3 ataxia telangiectasia (AT) patients, 2 obligate AT heterozygotes, 7 patients with diseases characterized by cellular hypersensitivity to ultraviolet radiation (UV), and 10 Duchenne muscular dystrophy (DMD) patients.

Recovery of absolute threshold with UVA-induced retinal damage.

A within-trial psychophysical procedure tracked the initial loss and subsequent recovery of visual thresholds in albino rats exposed to ultraviolet light at 350 nanometers and 0.4 milliwatts per square centimeter. Absolute thresholds increased up to 5 log units immediately following the 15 hour ultraviolet exposure, with a daily recovery of 1-2 log to asymptotic thresholds over a 7-day post-exposure period. The corresponding retinal damage on Day 1 included extensive vesiculation of the photoreceptor outer segments, vacuolation of the inner segments, and pyknosis of cell nuclei. The total number of photoreceptor nuclei and outer segments was unchanged relative to control eyes through post-exposure Day 3. Both nuclei and outer segment counts then consistently decreased 15-20 percent between Days 3-7. The two-stage loss of photoreceptors but daily recovery of absolute thresholds again suggests a significant dissociation of retinal structure and psychophysical function in light-induced ocular pathology.