Measurement of Critical Structures around Paraclinoidal Area : A Cadaveric Morphometric Study

OBJECTIVE: Although removal of the anterior clinoid process (ACP) is essential surgical technique, studies about quantitative measurements of the space broadening by the anterior clinoidectomy are rare. The purposes of this study are to investigate the dimension of the ACP, to quantify the improved exposure of the parasellar space after extradural anterior clinoidectomy and to measure the correlation of each structure around the paraclinoidal area. METHODS: Eleven formalin-fixed Korean adult cadaveric heads were used and frontotemporal craniotomies were done bilaterally. The length of C6 segment of the internal carotid artery on its lateral and medial side and optic nerve length were checked before and after anterior clinoidectomy. The basal width and height of the ACP were measured. The relationships among the paraclinoidal structures were assessed. The origin and projection of the ophthalmic artery (OA) were investigated. RESULTS: The mean values of intradural basal width and height of the ACP were 10.82 mm and 7.61 mm respectively. The mean length of the C6 lateral and medial side increased 49%. The mean length of optic nerve increased 97%. At the parasellar area, the lengths from the optic strut to the falciform liament, distal dural ring, origin of OA were 6.69 mm, 9.36 mm and 5.99 mm, respectively. The distance between CN III and IV was 11.06 mm. CONCLUSION: With the removal of ACP, exposure of the C6 segments and optic nerve can expand 49% and 97%, respectively. This technique should be among a surgeon's essential skills for treating lesions around the parasellar area.

Expression of Intermediate Filament Proteins in Rat Brain after Cryoinjury / 대한소아신경학회지

PURPOSE: Traumatic brain injury is a multifaceted injury that involves direct mechanical damage, intraparenchymal and subarachnoid hemorrhage, breakdown of the blood-brain barrier, excitotoxicity, and ischemia. Even though much investigations were performed, acceptable mechanical informations were rare. The aim of this study was to reveal the expression pattern of intermediate filament proteins associated with gliotic scars in cerebral cortex of rats after cryoinjury. METHODS: A total of 18 male Sprague-Dawley rats weighing 300 g, 2 months old, were used throughout the experiments. To injure the brain, rats were anesthetized for surgery with 3.5% chloral hydrate(1 mL/100 g, intraperitoneally); the frontal bones were exposed by elevating the skin; and craniectomies were performed adjacent to the central suture, midway between lambda and bregma. A cryoinjury was then created by applying a cold probe(3-mm-diameter steel rod chilled in liquid nitrogen) to the left frontal cortex(ipsilateral cortex) for 1 min. Rats were sacrificed at 1, 4, 7 and 14 days postsurgery(n=3, per time point), and three rats were sacrificed as normal controls. Serial brain cryosections were made by cryostat. For immunohistochemistry, brain tissue sections were allowed to react with mouse anti-rat GFAP antibody(1:200), mouse anti-rat vimentin antibody(1: 200), and mouse anti-rat nestin antibody(1:200). RESULTS:Reactive astrocytes expressing GFAP, vimentin and nestin appeared for the first time at 6 hours after cryoinjury. Proliferation of GFAP and nestin positive cells started at 1 day after cryoinjury, reached its maximum on day 4, and returned to normal level after the 7th post-injured day. Proliferation of vimentin positive cells started at 1 day after cryoinjury, reached its maximum on day 4, and returned to normal level after the 14th post-injured day. Characteristic morphological changes in reactive astrocytes were seen at 4 days after cryoinjury. CONCLUSION: The above results suggest that GFAP, vimentin and nestin positive cells attend in the formation of gliotic scars.

Effect of Exercise Training on Histochemical Muscle Recovery after Sciatic Nerve Crush in Rat / 체질인류학회지

It is constant controversy that exercise influence muscle regeneration in peripheral neuropathy. The aim of this experiment is to show that treadmill running exercise under well-controlled conditions is to improve of regeneration in rat gastrocnemius muscles after sciatic nerve crushing injury. Male Sprague-Dawley rats (1 month old, weight 150~180 g) were submitted to bouts of exercise on a treadmill up a 10 degrees decline and speed is 20 m/min for 60 min per day and gastrocnemius muscles were analysed at different exercise periods (5, 10, 15, 20 and 40 days) by immunohistochemistry in comparison with injured non-exercised muscles. Rats were sacrificed at 12th (5 days exercise), 19th (10 days exercise), 26th (15 days exercise), 33rd day (20 days exercise), 61st day (40 days exercise) after sciatic nerve crushing injury. It showed that type II myofibers (target fibers) on center area had reinnervation at sciatic nerve crush injury at 26th day in exercise rats, as at 33rd day appeared giant type II myofibers, myofibers grouping observed in regenerative muscle character, component ratio of closed normal muscle showed at 61st day. Giant type II myofibers showed at 33rd day in non-exercise rats, however did not nearly normal muscle at 61st day. Therefore we concluded that treadmill running exercise is able to improve regeneration processes in gastrocnemius muscles after sciatic nerve crushing injury of rats.

Antigen Presenting Cells Appearing in Injured Rat Brain Detected by Immunohistochemistry Method / 대한소아신경학회지

PURPOSE: Traumatic brain injury is a multifaceted injury that involves direct mechanical damage, intraparenchymal and subarachnoid hemorrhage, breakdown of the blood- brain barrier, excitotoxicity, and ischemia. Despite the dozens of previous investigations, the information about its pathogenic mechanism is still limited. The aim of this study was to reveal the appearance of antigen presenting cells in the cerebral cortex of rats after cauterization. METHODS: A total of 18 male Sprague-Dawley rats weighing 300 g and 2 months old on the average were used throughout the experiment. The frontal bones were exposed by elevating the skin and craniectomies were performed adjacent to the central suture, midway between lambda and bregma. Cauterizing injury was then created by battery-operated small vessel cauterizers to the left frontal cortex. The rats were sacrificed on the 1st, 4th, 7th and 14th days after the surgery(n=3, each time), and three rats were sacrificed as normal controls. Serial brain cryosections were made by cryostat. For immunohistochemistry, brain tissue sections were allowed to react with mouse anti-rat MHC class II antibody(1:500) and mouse anti-rat ED2 antibody(1:200). Also, brain tissues were routinely stained by H-E, and then microscopic observation and cell counts were performed. RESULTS: 1) MHC class II positive dendritic cells were absent in normal cerebral cortex parenchyme, but were found 28 times more in number in injured rats on the 7th day after cauterization. 2) ED2 positive macrophages were absent in normal cerebral cortex parenchyme, and were found 16 times more in number in injured rats on the 7th day after cauterization. 3) The number of MHC class II positive dendritic cells were smaller in number than that of ED2 positive macrophages 6 hours and 1st day later after cauterization, but it was higher in number on the 4th, 7th and 14th days. 4) The number of MHC class II positive dendritic cells were higher in number than that of ED2 positive macrophages around blood vessels and peripheral regions in the injured brain. 5) MHC class II positive dendritic cells were usually aggregated. CONCLUSION: It can be suggested that the increase in number of two kinds of antigen- presenting cells affect cell-mediated immune responses and phagocytosis.

Age-related Changes of MHC Class II Positive Dendritic Cells in Cardiac Muscle of Rat / 체질인류학회지

Cardiac dendritic cells are considered to play an important role in the immunoresponse of the heart. However, It is unknown that changes of shapes and numbers of these cells in the heart. The aim of this study is to reveal age-related changes of MHC class II positive dendritic cells in cardiac muscle of rat. Male Sprague-Dawley rats (1 month, 12 months, and 24 months old) were used in this study. Animals were deeply anesthetized with 3.5% chloral hydrate (1 mL/100 g) and hearts removed. Immunostaining was done according to standard methods used routinely. In brief, tissue sections were incubated with primary antibodies generated in mouse anti-rat MHC class II antibody for single immunostains. Tissue sections were observed by using light microscope and dendritic cells were counted. Average numbers of MHC class II-positive dendritic cells were 1.4 cells per unit area (0.2 mm2) at 1 month old rat, 2.8 cells at 12 months old rat, and 4.6 cells at 24 months old rat, and then numbers of dendritic cells were increased according to ages. According as age increases, cytoplasmic processes of MHC class II-immunoreactive dendritic cells became longer and more complex and aggregated together. It's suggested that age-related changes of MHC class II positive dendritic cells in the cardiac muscle would be related to immunity.

Age-related Changes of Antigen Presenting Cells in Rat Brain / 체질인류학회지

Activation of T cells for an immune response requires the participation of antigen presenting cells that express class II major histocompatibility complex gene products on their surface. As far as we know, there is no study on the agerelated changes of ED2 immunoreactive macrophages and MHC class II immunoreactive dendritic cells in the normal rat brain. The aim of the present study is to investigate the age-related changes of dendritic cells and macrophages in rat brain. The distribution and morphology of the macrophages and dendritic cells in the rat brain were studied from the 1 month-, 12 month- and 24 month-old rats by means of immunohistochemical methods using anti-rat MHC class II and anti-rat ED2 monoclonal antibodies. Antigen presenting cells were observed in choroid plexuses and white matter of the rat brain. The numbers of antigen presenting cells gradually increased with age. At all age stages and regions of the rat brain, the numbers of ED2 immunoreactive macrophages was higher than that of MHC class II immunoreactive dendritic cells. According as age increases, shapes of antigen presenting cells became more complex and aggregated together. In conclusion, the above results suggest that the increases of the number and the changes of the morphology in two kinds of the antigen-presenting cells, MHC class II-immunoreactive dendritic cells and ED2-immunoreactive macrophages, with age may influence on effects of cell-mediated immune responses.

Effect of Exercise to Improve of Rat Lower Limb Healing after Physical Injury / 체질인류학회지

The aim of this report is to show that treadmill running exercise under well-controlled conditions is to improve of regeneration in rat gastrocnemius muscles after physical injury. For this, rats were submitted to bouts of exercise on a treadmill up a 10 degrees decline for 60 min and gastrocnemius muscles were analysed at different exercise periods by immunohistochemistry in comparison with injured nonexercised muscles. Rats were used with guidelines for experimental procedures as set forth in the Declaration of Helsinki. We analysed the regenerative processes by detection of immunoreactivity for the two intermediate filaments, desmin and vimentin. Desmin and vimentin are specific components of the cytoskeleton of striated muscle fibers and of mononuclear cells of mesenchymal origin including myoblasts, respectively. We found that non-exercised rats had more desmin-and vimentin-positive myofibers than that of exercised rats at 9th, 16th, 23th, 30th day after physical injury. At 30th day, non-exercised rats had several desmin-and vimentinpositive myofibers, but exercised rats had numerous normal myofibers. These results show that exercise is able to improve regeneration processes in physical injured gastrocnemius muscles of rats.

Changes of c-Fos Immunoreactivity in Midbrain by Deep Pain and Effects of Aspirin

PURPOSE: It had been suggested that pain arising from deep somatic body regions influences neural activity within periaqueductal gray(PAG) of midbrain via distinct spinal pathways. Aspirin is one of the popular non-steroidal anti-inflammatory drugs used in the management of pain. Fos expression was used as a marker for neuronal activity throughout central neurons following painful peripheral stimulation. This study was prepared to investigate changes of c-Fos immunoreactivity in midbrain by deep pain and effects of aspirin. METHODS: Male Sprague-Dawley rats were injected with 0.1 mL of 5% formalin in the plantar muscle of the right hindpaw. For experimental group II, aspirin was injected intravenously before injection of formalin. An aspirin-untreated group was utilized as group I. Rats were sacrificed at 0.5, 1, 2, 6 and 24 hours after formalin injection. Rat's brains were removed and sliced in rat brain matrix. Brain slices were coronally sectioned at interaural 1.00-1.36 mm. Serial sections were immunohistochemically reacted with polyclonal c-Fos antibody. The numbers of c-Fos protein immunoreactive neurons in ventrolateral periaqueductal gray(VLPAG) and dorsomedial periaqueductal gray(DMPAG) were counted and analyzed statistically with Mann-Whitney U tests. RESULTS: Higher numbers of c-Fos protein immunoreactive neurons were found in VLPAG. In both VLPAG and DMPAG of formalin-treated group, the numbers of c-Fos protein immunoreactive neurons were significantly higher at all time points than the formalin-untreated group, which peaked at two hours. The numbers of c-Fos immunoreactive neuron of the aspirin-treated group were less compared to the aspirin-untreated group at each time point. CONCLUSION: These results provide some basic knowledge in understanding the mechanism of formalin-induced deep somatic pain and the effects of aspirin.

Age-Related Changes of MHC Class II-immunoreactive Dendritic Cells in Rat Brain / 대한소아신경학회지

PURPOSE: Dendritic cells are antigen presenting cells(APC) that express class II major histocompatibility complex gene products on their surface. Recently, it was proved that dendritic cells activate antitumor immunity for intracranial germ cell tumor. The aim of the present study is to investigate the age-related changes of MHC class II-immunoreactive dendritic cells in the rat brain. METHODS: Male rats(Sprague-Dawley) were sacrificed at 1 month, 12 months and 24 months after birth. Brains were removed and sliced in rat brain matrix. Brain slices were cryosectioned coronally at interaural 5.70-6.70 mm. Brain tissue sections were immunohistochemically reacted with monoclonal MHC class II antibody. RESULTS: MHC class II-immunoreactive dendritic cells were observed in choroid plexuses and white matter(corpus callosum, cerebral peduncle and external capsule). The number of MHC class II-immunoreactive dendritic cells was slightly increased with age. As age increases, shapes of MHC class II-immunoreactive dendritic cells became more complex and aggregated together. CONCLUSION: As age increases, MHC class II-immunoreactive dendritic cells in choroid plexuses and white matter of the brain became not only more complex in shape, but also increased in number to improve immunity.

Inflammatory Response of Secretoneurin -Immunoreactive Nerves in the Rat Uterus after Formalin Exposure / 체질인류학회지

It is known that there are numerous chemotatic secretoneurin -immunoreactive nerve fibers and movable MHC class II -immunoreactive dendritic cells in the normal uterine cervix. And the relationships between them are not fully understood. The aim of this study is to reveal that secretoneurin could give to chemotatic influence to dendritic cells in inflammational state. Virgin female Sprague -Dawley rats (n = 20; approximately 2 months old; 200 ~250 g body weight) were used in this study. Animals (n = 10) were injected with 5% formalin (0.5 ml/day, 5 days) in experiment group. Animals were deeply anesthetized with 3.5% chloral hydrate (100 mg/kg, i.p.) and uterine cervix were removed. Immunostaining was done according to standard methods used routinely. In brief, tissue sections were incubated with primary antibodies generated in mouse anti -rat MHC class II antibody and mouse or rabbit anti -rat secretoneurin antibody for single and double immunostains. FITC for secretoneurin and rhodamine for MHC class II were used as secondary antibodies in double stains. Tissue sections were observed by using light and confocal laser scanning microscopes. The results were as follows; 1. Numerous secretoneurin -immunoreactive nerve fibers were located in the lamina propria and those were not found in the epithelium of normal rat uterine cervix. 2. MHC class II -immunoreactive dendritic cells were mainly located in the epithelium and the lamina propria of normal rat uterine cervix. 3. On the inflammation state, MHC class II -immunoreactive dendritic cells were mainly located in the lamina propria and those were not found in the epithelium of rat uterine cervix. According to above results, it is suggested that secretoneurin can give to chemotatic influence to dendritic cells in inflammational state. Therefore, secretoneurin is considered to be used for dendritic cell immunotheraphy.

Phenotypic Changes of Intermediate Filament Proteins during Degeneration and Regeneration of Rat Leg Muscles following Physical Injury / 대한정형외과학회잡지

PURPOSE: The aim of this experiment was to observe the phenotypic changes of intermediate filaments in skeletal muscle fibers during the degeneration and regeneration of the physical injury. MATERIALS AND METHODS: The gastrocnemius muscles of rats were physically damaged by needles and serial cryosections of the damaged muscles were prepared at 2, 4, 6, 9, 15, 21 and 35 days after injury. The cryosections were immunolabelled with desmin, vimentin and histochemically reacted with NADH-TR (nicotinamide adenine dinucleotide tetrazolium reductase). RESULTS: Myotubes, asvisualized by desmin and vimentin, appeared at 9 days after injury. The regenerative myofibers were similar to normal muscles 35 days after injury. Degeneration and regeneration occurred simultaneously and positive reactions for desmin disappeared earlier than those of vimentin. CONCLUSION: Both desmin and vimentin are strong staining tools for the evaluation of myopathy. The phenotypic patterns of intermediate filaments showed various degrees of regeneration in the early stages after physical injury.

Immunocytochemical Study on Distribution and Morphology of Dendritic Cells in the Tongue during Development and Growth of the Rat / 체질인류학회지

Activation of T cells for an immune response requires the participation of antigen presenting cells (APC) that express class II major histocompatibility complex gene products on their surface. Until recently, the macrophages have been considered to be the prime candidates for this role, but it is now recognized that other cells, including dendritic cells, B cells, activated T cells and endothelial cells, can present antigen effectively. Particularly, among them, dendritic cells (DC) are considered to be very efficient APC for various T -cell dependent immune responses in comparison with other types of APC. Nonlymphoid dendritic cells including Langerhans cells and interstitial dendritic cells strongly express the MHC class II products and have characteristic dendritic morphology. As far as we know, there is no study on the ontogeny of MHC class II -immunoreactive dendritic cells in the rat tongue. The aim of the present study is to investigate the ontogeny and morphological characterization of dendritic cells in the tongue of growing and developing rats. The distribution and morphology of the dendritic cells in the rat tongue were studied from the fetal 15 -day until 180 days after birth by means of immunocytochemical methods using anti -rat MHC class II monoclonal antibodies. The results were as follows: 1. MHC class II -immunoreactive dendritic cells were first found in the muscle layer of 17 -day fetus, and in the epithelium and lamina propria of the tongue at birth. 2. The number of MHC class II -immunoreactive dendritic cells was gradually increased with age, particularly, in the epithelium at 14th day, in the lamina propria at 14th day, and in the muscle layer at 21st day after birth. 3. Numbers of MHC class II -immunoreactive dendritic cells were higher in the dorsal part than in ventral part of the tongue at all developmental stages. Especially, dendritic cells were twice higher numbers in the epithelium, a little higher numbers in the lamina propria and almost same numbers in the muscle layer. 4. With age, shapes of MHC class II -immunoreactive dendritic cells were changed from round to dendritic and aggregated together. In conclusion, the above results suggest that the increases of the number and the changes of the morphology in MHC class II -immunoreactive dendritic cells, with age may influence on effects of cell -mediated immune responses.

Effects of c -Fos Immunoreactivity on Central Nervous System after Administration of Aspirin / 체질인류학회지

Aspirin is one of the popular non -steroid anti -inflammatory drugs used in the management of pain. This study was performed to investigate the effects of aspirin on c -Fos expression in rat CNS after inducing somatic pain with formalin. Male S.D. rats were injected subcutaneously with 0.1 ml of 5% formalin in the plantar surface of right hindpaw. For experimental group, aspirin was administered orally before injection of formalin. Asprin -untreated group was utilized as the control group. Rats were sacrificed at 0.5, 1, 2, 6 and 24 hours after formalin injection. Rat brains were removed and sliced in rat brain matrix. Brain slices were coronally sectioned at interaural 5.70 ~6.70 mm. Serial sections were immunohisto-chemically reacted with polyclonal c -Fos antibody. The numbers of c -Fos protein immunoreactive neurons in the cingulate cortex, primary somatosensory area, and hippocampus were counted and analyzed statistically with Mann - Whitney U test. Results were as follows: 1. Higher numbers of c -Fos immunoreactive neurons were found in the cingulate cortex, primary somatosensory area and hippocampus. 2. Both aspirin -treated and -untreated groups, numbers of c -Fos immunoreactive neurons were significantly higher all time points than formalin -untreated group, which peacked at 2 hours. 3. The numbers of c -Fos immunoreactive neuron of the aspirin -treated group were less compared to the aspirin - untreated group at each time point. In conclusion, these results provide some basic knowledge in understanding the mechanism and control of formalin - induced somatic pain.

Expression Changes of c-Fos Protein of Rat Brain Following Pentylenetetrazol-induced Seizures / 대한소아신경학회지

PURPOSE: The expression of c-Fos protein has been shown to be a useful marker for elevated levels of neuronal activity generated in the brain following different stimuli, including seizures. This study was conducted to investigate distribution and numbers of neurons where dentate and cingulate gyrus become activated following pentylenetetrazol-induced seizures by means of expression patterns of c-Fos protein. METHODS: Rats were sacrificed at increasing times(1 hour, 2 hours, 8 hours, 1 day, 4 days and 7 days) after pentylenetetrazol-induced seizure. Rat brains were removed and sliced in rat brain matrix. Brain slices were coronal sectioned at interaural 5.70-6.70mm. Serial sections were immunohistochemically reacted with polyclonal c-Fos antibody. The distribution and numbers of c-Fos protein immunoreactive neurons in dentate gyrus and cingulate gyrus were examined and analyzed statistically with Mann-Whitney U test. RESULTS: The numbers of c-Fos protein immunoreactive neurons in dentate gyrus peaked at 1 hours and reached almost normal conditions at 7 days after seizure. Also, same patterns were occurred in cingulate gyrus. Concentration value that pentylenetetrazol can induce was different from each animals and c-Fos immunoreactive cells were various kinds of neurons. CONCLUSION: Higher numbers of c-Fos protein immunoreactive neurons were found in dentate and cingulate gyrus at the same times after seizure. These findings suggest that neurons of dentate and cingulate gyrus play a crucial role in seizure onset following pentylenetetrazol-induced seizure.

Changes of c-Fos Immunoreactivity in Rat Brain Neurons According to Ages After Induced Pain / 대한소아신경학회지

PURPOSE: The pathways of pain conduction in brain are not well known. Also, differences of somatic pain conduction between adult and young age have not been fully elucidated. This study was conducted to investigate any differences in the expression of c-Fos protein between adult and young rats after somatic pain was induced by formalin. METHODS: Male rats (n=70) were injected subcutaneously with 0.1mL of 5% formalin in the plantar surface of right hindpaw. Rats were sacrificed at 30 minutes, 1 hour, 2 hours, 6 hours, 24 hours after noxious formalin stimuli to hindpaws and rectums. Rat brains were removed and sliced in rat brain matrix. Brain slices were coronally sectioned at interaural 5.70-6.70mm. Serial sections were immunohistochemically reacted with polyclonal c-Fos antibody. The numbers of c-Fos protein immunoreactive neurons in cingulate cortex, primary somatosensory area, and hippocampus were examined and analyzed statistically with Mann-Whitney U test. RESULTS: The number of c-Fos protein immunoreactive neurons in the cingulate cortex, primary somatosensory area and hippocampus peaked at 2 hours after formalin-induced pain on adult rats. The number of c-Fos protein immunoreactive neurons in the cingulate cortex, primary somatosensory area and hippocampus peaked at 1 hour after formalin-induced pain on young rats. The numbers of c-Fos protein immunoreactive neurons of adult groups were higher than that of young groups at all points of time. CONCLUSION: The immunoreactions in adult group expressed more intense than those in young group. Earlier expression of immunoreactions in young group suggests of faster conduction of pain, compared to those in adult group. Larger number of c-Fos protein immunoreactive neurons were found within specific regions in both groups. These results could provide some basic knowledge in understanding the mechanism and control of pain in pediatric group.

Phenotypic Changes of Intermediate Filament Proteins during Degeneration and Regeneration of Rat Leg Muscles following Sciatic Nerve Injury / 대한정형외과연구학회지

PURPOSE: The intermediate filament proteins, desmin and vimentin, are specific components of the cytoskeleton of striated muscle fibers and of mononuclear cells of mesenchymal origin including myoblasts, respectively. Desmin has also been found in presumptive myoblasts of mammals. The aim of this experiment was attempted to observe the phenotypic changes of intermediate filaments in skeletal muscle fibers during early stages of sciatic nerve crushing injury. MATERIALS AND METHODS: The sciatic nerves of rats were surgically crushed by hemostat and serial cryosections of soleus and extensor digitorum longus(EDL) muscles were prepared at 2, 4, 6, 8, 10, 15, 20 and 27 days after nerve injury. Serial cryosections were immunolabelled with desmin, vimentin and laminin and were histochemically reacted with NADH-TR. RESULTS: 1) Firstly, desmin positive fibers were appeared in fast-twitch type C fibers of both muscles at 6 days after nerve crushing, but were not reacted for vimentin. 2) Co-expressions of desmin and vimentin were firstly detected in fast-twitch type A fibers of EDL muscles at 8 days after nerve injury. In soleus muscles, co-expressions of desmin and vimentin were firstly seen in slow-twitch type B fibers at 10 days after nerve injury. Many atrophic fibers, that contained several central nuclei like myotubes and co-expressed desmin and vimentin, were appeared in EDL muscles at 10 days after nerve injury. Although whole regions of fibers were regenerated in EDL muscles, only peripheral regions of fibers were regenerated in soleus muscles at 15 days after nerve injury. Many atrophic fibers, co-expressed of desmin and vimentin, were appeared in EDL muscles at 20 days after nerve injury. These whole fibers represented various degrees of regenerating stages. Most of mature fibers containing several central nuclei, only expressed vimentin slightly, were seen in soleus muscles at 20 days after nerve injury. Most fibers of both muscles were matured at 27 days after nerve injury, but some fibers in EDL muscles were still in processing of degeneration and regeneration. No expressions of desmin and vimentin indicated that muscle fibers were almostly matured in soleus muscles at 27 days after nerve injury. 3) Targetoid or target fibers which informed reinnervation, were appeared firstly in soleus muscles at 20 days and were seen in both muscles at 27 days after nerve injury. All targetoid and target fibers were type B fibers. CONCLUSION: Desmin was revealed in processes of degeneration and regeneration and vimentin was appealed in regeneration process. At the same time, positive immunoreactivity of desmin and vimentin showed specific differences in degree of degeneration and regeneration according to different muscles and muscle fibers.

Phenotypic Changes of Intermediate Filament Proteins during Degeneration and Regeneration of Rat Leg Muscles following Sciatic Nerve Injury / 대한정형외과연구학회지

PURPOSE: The intermediate filament proteins, desmin and vimentin, are specific components of the cytoskeleton of striated muscle fibers and of mononuclear cells of mesenchymal origin including myoblasts, respectively. Desmin has also been found in presumptive myoblasts of mammals. The aim of this experiment was attempted to observe the phenotypic changes of intermediate filaments in skeletal muscle fibers during early stages of sciatic nerve crushing injury. MATERIALS AND METHODS: The sciatic nerves of rats were surgically crushed by hemostat and serial cryosections of soleus and extensor digitorum longus(EDL) muscles were prepared at 2, 4, 6, 8, 10, 15, 20 and 27 days after nerve injury. Serial cryosections were immunolabelled with desmin, vimentin and laminin and were histochemically reacted with NADH-TR. RESULTS: 1) Firstly, desmin positive fibers were appeared in fast-twitch type C fibers of both muscles at 6 days after nerve crushing, but were not reacted for vimentin. 2) Co-expressions of desmin and vimentin were firstly detected in fast-twitch type A fibers of EDL muscles at 8 days after nerve injury. In soleus muscles, co-expressions of desmin and vimentin were firstly seen in slow-twitch type B fibers at 10 days after nerve injury. Many atrophic fibers, that contained several central nuclei like myotubes and co-expressed desmin and vimentin, were appeared in EDL muscles at 10 days after nerve injury. Although whole regions of fibers were regenerated in EDL muscles, only peripheral regions of fibers were regenerated in soleus muscles at 15 days after nerve injury. Many atrophic fibers, co-expressed of desmin and vimentin, were appeared in EDL muscles at 20 days after nerve injury. These whole fibers represented various degrees of regenerating stages. Most of mature fibers containing several central nuclei, only expressed vimentin slightly, were seen in soleus muscles at 20 days after nerve injury. Most fibers of both muscles were matured at 27 days after nerve injury, but some fibers in EDL muscles were still in processing of degeneration and regeneration. No expressions of desmin and vimentin indicated that muscle fibers were almostly matured in soleus muscles at 27 days after nerve injury. 3) Targetoid or target fibers which informed reinnervation, were appeared firstly in soleus muscles at 20 days and were seen in both muscles at 27 days after nerve injury. All targetoid and target fibers were type B fibers. CONCLUSION: Desmin was revealed in processes of degeneration and regeneration and vimentin was appealed in regeneration process. At the same time, positive immunoreactivity of desmin and vimentin showed specific differences in degree of degeneration and regeneration according to different muscles and muscle fibers.

Changes of c-Fos Protein Expression in Rat Brain Neurons after Formalin Induced Pain / 대한소아신경학회지

PURPOSE: The effects of pain on brain is not well known. Also, differences between somatic and visceral pains have not been fully elucidated. This study was conducted to investigate changes in the expression of c-Fos protein after somatic and visceral pains were induced by formalin. METHODS: Male rats(n=65) were underwent one of three procedures : (i) Control group, rats were left undisturbed in their cages; (ii) Somatic pain group, rats were injected subcutaneously with 0.1 ml of 10% formalin in the plantar surface of right hindpaw; (iii) Visceral pain group, rats were administered with same amount of formalin, as described above, in the rectum. Rats were sacrificed at increasing times(30 minutes, 1 hour, 2 hours, 6 hours, 1 day, 3 days and 7 days) after noxious formalin stimuli to hindpaws and rectums. Rat brains were removed and sliced in rat brain matrix. Brain slices were coronal sectioned at interaural 5.70-6.70mm. Serial sections were immunohistochemically reacted with polyclonal c-Fos antibody. The numbers of c-Fos protein immunoreactive neurons in cingulate cortex, primary somatosensory area, and hippocampus were examined and analyzed statistically with Mann-Whitney U test. RESULTS: 1) The numbers of c-For protein immunoreactive neurons in cingulate cortex, primary somatosensory area and hippocampus peaked at 2 hours after somatic pain stimuli and reached almost normal conditions at 7 days. 2) The numbers of c-Fos protein immunoreactive neurons in cingulate cortex, primary somatosensory area and hippocampus peaked at 1 day after visceral pain stimuli and reached almost normal conditions at 7 days. 3) The numbers of c-Fos protein immunoreactive neurons of somatic pain groups were higher than that of visceral groups at all times and the difference of numbers peaked at 2 hours after pain stimuli. CONCLUSION: Reactions of somatic pain stimuli influenced more changable than visceral pain stimuli to brain. Conduction velocities of somatic pain were more faster than those of visceral pain. Higher numbers of c-Fos protein immunoreactive neurons were found in specific regions. These results provide some basic knowledge in understanding the mechanism and control of pain.

A Study on the Epidermis Undergoing Apoptosis After Bone Marrow Transplantation / 대한해부학회지

GVHD (Graft-versus-Host Disease) results from the cytotoxic T lymphocytes from the bone marrow recognizing the recipient's minor histocompatibility antigens. In experimental murine models, either CD4+ or CD8+ T-cell subsets can cause GVHD, depending upon the particular strain combination utilized. Recent studies suggest that the keratinocyte undergo apoptosis in GVHD. However, morphological data supporting this concept are still lacking. The present study was undertaken in order to document apoptosis in experimental acute GVHD via sequential analysis of ultrastructure .Acute GVHD was produced across minor histocompatibility loci using appropriately matched murine strains. Acute GVHD was mediated with the use of highly purified preparations of donor CD4+ and CD8+ T-cell subsets. Whole T cells were used as a positive control and T cell depleted bone marrow as a negative control. Conventional transmission electron microscopy was used to define apoptosis structurally Sequential ultrastructure revealed that the keratinocyte underwent apoptosis in CD4+, CD8+ and whole T cell groups. This study demonstrates the sequential ultrastructure of the keratinocyte undergoing apoptosis from the beginning to the end. Both of the basal and the suprabasal keratinocytes show the morphology of early apoptosis, and the detachment of the tonofibril from the basement membrane and the adjacent cell was the general findings in the apoptotic cell Sequences of the cytoplasmic condensation was demonstrated . Through ultrastructural quantitation the apoptotic indices were depicted in all the experimental groups. Characteristically, numerous lymphocytes underwent apoptosis in CD8+ groups at day 28 and 35.

The Differential Effects of Cyclophosphamide on Dendritic Cells and Tissue Macrophages / 대한해부학회지

This study was undertaken to investigate the effects of cyclophosphamide (CY) on dendritic cells (DCs) and ED2 positive tissue macrophages in adult rat lymphoid and non-lymphoid organs. A single dose of CY (150 mg/kg) was administered to Sprague-Dawley rats by intraperitoneal injection. After the rats were sacrificed in groups of three at 1 day, 3 days, 1 week, 2 weeks and 1 month, the tongue, heart, urinary bladder, thymus, spleen and cervical lymph nodes were removed. The immunocytochemical characterization of the cells was carried out using the monoclonal antibodies OX6, OX62 and ED2 in cryostat-cut sections. CY elicited a decrease in number of intraepithelial and interstitial DCs in urinary bladder and in number of interdigitating DCs in all the lymphoid organs examined in this study, whereas CY did not cause any alteration in the number and distribution pattern of dendritic cells in tongue and hearts. CY increased the size and number of tissue macrophages in all the organs examined in this study. Most of these features began to appear from the first day and reached the maximun on the third and seventh day, but two weeks after CY administration, these phenomena declined. In conclusion, CY has differential effects on the rat DCs and macrophages and also on the subpopulations of DC according to the location and the functional state.