• International Journal of Oral Biology
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• v.43 no.2
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• pp.93-100
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• 2018

The investigation of the embryonic development of the cerebellum has a long history. The postnatal normal development of the cerebellum in rodents and other animals became a popular topic for morphological investigations nearly a century ago. However, surprisingly, only a few studies are available regarding the prenatal normal development of the rodent cerebellum, especially in guinea pigs. Cell proliferation is essential for the development of the nervous system. The assessment of cell proliferation can be achieved by using various methods. In this study, we investigated the cell proliferation of the cerebellar cortex in guinea pigs at different stages of pregnancy and in postnatal life. Fetuses were obtained by cesarean section at 50 or 60 days of gestation (dg). Immunohistochemistry was performed with proliferating cell nuclear antigen (PCNA) antibody in the cerebellum. Strong PCNA immunoreactivity was observed in the external granular layer (EGL), which is a neurogenic zone in the cerebellum. The proportion of PCNA-IR cells was greater at 1 week than at 60 dg in lobule I, but not lobule VIII. After 50 dg, the width of the EGL continued to decline until 1 week, due to the maturation of the EGL cells. These results demonstrate the pattern of PCNA immunoreactivity in the developing cerebellum of guinea pigs. This serves as a guideline to study abnormal cerebellum development.

• Journal of Life Science
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• v.14 no.6 s.67
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• pp.975-985
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• 2004

This study attempts to investigate the developmental alterations of rat cerebral cortex, and the effects of EGEE on the developmental cerebral cortex in the prenatal, postnatal and adults were examined by morphological methods and H-E staining was used for the histological changes. In the case of injection of EGEE, at 14 day of fetal phase, parietal cortex was thickest $(95{\pm}12.7\;{\mu}m)$ but, it was thinner than in the control group $(102{\pm}14.0\;{\mu}m)$ and, occipital cortex $(57{\pm}10.5\;{\mu}m)$ compared with other cortexes was the thinnest in fetal phase. In the suckling phase, each cortex grew thick quickly but, after weanning phase, the growth of the cortex slowed and the thickness of cortex was similar to that of cortex in the adult phase. At 105 day after birth, the parietal cortex was thickest $(934{\pm}21.6\;{\mu}m)$ but, decreased compared with control group $(1113{\pm}19.0\;{\mu}m)$. When EGEE was injected in intraperitoneal of rat, the number of neuroblasts per unit area was largest $(207.7{\pm}11.4/10^{-2}\;mm$ at the mantle layer of parietal cortex at 14 day of fetal phase but, decreased compared with control group $(224.2{\pm}13.8/10^{-2}\;mm$ , and the size was largest $(7.5{\pm}1.3\;{\mu}m)$ at the ependymal cell layer of occipital cortex at 3 day after birth but, decreased compared with control group $(9.0{\pm}1.2\;{\mu}m)$. Simillar to control group, the number of granular cells and pyramidal cells were largest at the II and III layer of parietal cortex, but decreased during developmental phase. The size was largest at the IV and V layer of occipital cortex but it was decreased compared with control group. When EGEE was injected in intraperitoneal of rat, the cerebral cortex from fetal phase to 3 day after birth has differentiated into the 3 layers; ependymal, mantle and marginal layer, but empty cisternaes or vacoules in the cerebral cortexes and the condensed phases of neuroblasts were appeared. From 5 day after birth, it has differentiated into the 4 layers; molecular, external granular, mixed layer of internal granular, external and internal pyramidal cells and multiformal layer but, empty cisternaes or vacoules in the granular and pyramidal cell layers were appeared and the number per unit area of neuron was decreased. In the cerebral cortex of the weaning and adult phases, division of cell layers was not clear and empty cisternae was formed in the cortex with the cells in external granular and pyramidal cell layers, was magnified or condensed around blood vessels of neurons.

• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.628-634
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• 1999

We have studied, by a nonisotopic in situ end-labeling(ISEL) technique, frequency of apoptosis in the external granular layer(EGL) of the cerebellum of immature mice by ${\gamma}$-rays irradiation from $^{60}Co$ or diagnostic ultrasound exposure. The total number of normal cells and cells showing morphological features of apoptosis were counted. The frequency of apoptotic cells was expressed as a percentage of the total number of cells in EGL. The extent of changes following 200 cGy(1090 cGy/min) was studied at 2, 4, 6, 8, 12, or 24 hours after exposure. The maximal frequency was found 6~8 hours after exposure. The immature mice that received 18, 36, 54, 108, 198, 396 cGy of ${\gamma}$-rays or diagnostic ultrasound(7.5MHz, 4.2mW, $I_{SPTA}=7.9mW/cm^2$, $I_{SPTA}=114.3W/cm^2$) for 10 or 30 minutes were examined 6 hours after irradiation. Measurements performed after ${\gamma}$-ray irradiation showed a dose-related increase in apoptotic cells in each of the mice studied. The dose-response curves were analyzed by a linear-quadratic model ; frequency of apoptotic cell in the EGL was y = $(0.1349{\pm}0.01175)D$+$(-0.0001522{\pm}0.0000334)D^2$+0.048($r^2$ = 0.981, D = dose in cGy). In the experiment of ultrasound exposure, the frequency of apoptotic cell was $0.106{\pm}0.130$(10 minutes exposure) and $0.167{\pm}0.220$(30 minutes exposure). We estimated the relative dose of the yield from the experiment with ultrasound by substituting the yield from ultrasound exposure into the curve from the ${\gamma}$-irradiation. The relative dose of ultrasound exposure compared with ${\gamma}$-irradiation were 0.432 cGy(10 minutes exposure) and 0.885 cGy(30 minutes exposure). We have found that there is no evidence to indicate that diagnostic ultrasound involves a significant risk.

• Development and Reproduction
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• v.15 no.3
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• pp.205-213
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• 2011

Ethanol treatment during the brain growth spurt period has been known to induce the death of Purkinje cells. The underlying molecular mechanisms and the role of reactive oxygen species (ROS) in triggering ethanol-induced Purkinje cell death are, however, largely unresolved. We undertook TUNEL staining, western blotting assay and immunohistochemistry for the cleaved forms of caspase-3 and -9, with calbindin D28K double immunostaining to identify apoptotic Purkinje cells. The possibility of ROS-induced Purkinje cell death was immunohistochemically determined by using anti-8-hydroxy-2'deoxyguanosine (8-OHdG), a specific cellular marker for oxidative damage. The results show that Purkinje cell death of PD 5 rat cerebellum following ethanol administration is mediated by the activation of caspase-3 and -9. However, unexpectedly, TUNEL staining did not reveal any positive Purkinje cells while there were some TUNEL-positive cells in the internal and external granular layer. 8-OHdG was detected in the Purkinje cell layers at 8 h, peaked at 12-24 h, but not at 30 h post-ethanol treatment. No 8-0HdG immunoreactive cells were detected in the internal and external granular layer. The lobule specific 8-OHdG staining patterns following ethanol exposure are consistent with that of ethanol-induced Purkinje cell loss. Thus, we suggest that ethanol-induced Purkinje cell death may not occur by the classical apoptotic pathway and oxidative damage is involved in ethanol-induced Purkinje cell death in the developing cerebellum.

• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.499-508
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• 1997

The distributions and morphological characteristics of neurons displaying immunoreactivity to the catecholamine synthetic enzymes, tyrosine hydroxylase(TH), dopamine-${\beta}$-hydroxylase(DBH), and phenylethanolamine-N-methyltransferase (PNMT) were examined in the adjacent sections of the olfactory bulb of the Striped Field Mouse(Apodemus agrarius coreae). None of these cell groups displayed either DBH or PNMT immunoreactivity. Many TH-immunoreactive neurons were present in the olfactory bulb. The vast majority of such cells occurred in the glomerular layer as periglomerular cells surrounding the glomeruli. Numerous addtional cells were present in the external plexiform layer, and scattered in the mitral cell layer and internal plexiform layer. Also TH-immunoreactive neurons were found in the glomerular layer and granular layer of the accessory olfactory bulb.

• Applied Microscopy
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• v.15 no.2
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• pp.31-68
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• 1985

Authors are trying to unveil the ultrastructural organization of olfactory bulb, which has been summerized under light microscopic level or communicated only in some detail in different view point until now. For the critical point of view, since the phylogenetical approach will give the ultimate value in the correlative study between structural and functional bases (Brodal, 1969), the present study was carried out light and electron microscopic analyses of the structures of the neurons and synaptic organizations in olfactory bulbs from different animals in phylogenetical scale. We selected each one species from five animal classes: the house rabbit(Oryctolagus cuniculus var. domesticus [Gmelin]) from Mammalia, the domestic fowl (Gallus gallus domesticus Brisson) from Aves, the viper (Agkistrodon hylys [G.P. Pallas]) from Reptilia, a frog (Bombiana orientalis Boulenger) from Amphibia and the crussian carp (Carassius carassius [Linne]) from Pisces. For light microscopic study, samples were fixed in 10% formalin and paraffin sections were stained with hematoxylin-eosin. For the electron microscopic study, the tissues were fixed by perfusion through the heart or immersion with 1% paraform-aldehyde-glutaraldehyde mixture (phosphate buffer, pH 7.4), and final tissue block trimmed under dissecting microscope were osmicated (1% OsO4), they were embedded in Araldite or Epon 812, and ultrathin sections were made by LKB-V ultratome following the inspection of semi-thin sections stained with toluidine blue-borax solution. Ultra-thin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100CX electron microscope. We have summerized our morphological analyses as follows: 1. The olfactory bulb of rabbit, viper and frog shows the eight layers of fila olfactoria, glomerular, external granular, external plexiform, mitral cell, internal plexiform, internal granular, medullary but domestic fowl shows the five layers of glomerular, fibrillar, mitral, granular and medullary and the three layers of fibrilla, glomerular and medullary in crussian carp. The sharpness of demarcation between the layers shows deferential tendency according to phylogenetical order. 2. Mitral cells of vertebrate have large triangular or oval shape with spherical nuclei which contain not so much chromatin. The cytoplasm contains numerous cell organelles, of which Nissl's bodies or granular endoplasmic reticula arranged as parallel strands. Development of granular endoplasmic reticula were declined as the phylogentical grade is going lower. 3. Tufted cells of all animal are mostly spindle or polygonal contour and contain oval nuclei which located in periphery of cytoplasm. The nuclei of rabbit, fowl, viper and frog has relatively space chromatin, but a nucleus of crussian carp contain irregularly aggregated chromatin in karyoplasm. Their cytoplasmic volume and cell organelle contents are in between those of mitral cell and granular cell. They contain moderate amount of mitochondria, granular endoplasmic reticula, a few Golgi complex, polysomes, lysosome, etc. 4. Granule of cells of all the vertebrate amimals studied exhibit similar features; cells and their dense nuclei show spherical or oval contour, and they have the thin rim of cytoplasm which contain only a few cell organelles. 5. In rabbit, the soma of mitral cells were in contact with boutons with two types of synaptic vesicles, that is, round and flat vesicles, especially flat vesicles in boutons were showing reciprocal synapses. However, in domestic fowls, vipers, frogs and crussian carps, there were found boutons showing only spherical synaptic vesicles. 6. The boutons containing round synaptic vesicles were made contact with the some of tufted cell of olfactory bulb in the rabbits, fowls, vipers and frogs, but no synaptic boutons were observed in soma of tufted cells in crussian carps. In the frogs, there were observed dendrites were contact with the soma of tufted cells. 7. In the neuropils of plexiform, granular and glomerular layers olfactory bulbs in the vertebrate, the synapses were axo-large dendrites, axo-median and small dendrites, dendrodendritic, and axo-axonal contacts. However, in the neuropil of crussian carps, synapses were observed only in glomerular layer.

• Archives of Reconstructive Microsurgery
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• v.24 no.1
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• pp.37-39
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• 2015

Epidermal inclusion cyst is a common mass in life. It is covered with a stratified squamous epithelium, thus, there is a granular cell layer adjacent to the keratin-containing cyst lumen. It can be caused by mechanical force, trauma, or a spontaneous event. It can rupture spontaneously or be ruptured by external mechanical forces. Epidermal inclusion cysts that exhibit inflammation or recur should be removed by simple excision. In this case, the patient showed an epidermal inclusion cyst under an anterolateral thigh free flap, which can cause the palpable mass to go unnoticed. First we thought he had neuroma formation after a surgical procedure on his foot. However it was an epidermal inclusion cyst, which was diagnosed by a special pathologist. It is a curious and rare case.

• Korean Journal of Veterinary Research
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• v.45 no.2
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• pp.139-149
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• 2005

In order to study about the lobule and layer formation and cell migration of the mouse cerebellum from at the birth to 15th day effected by 2.5, 5 and 10 Gy r-raddiation at the 19th pregnancy. The routine tissue preparation and staining procedure, Immunohistochemical staining method by the several antibody and western brotting method were utilized from the birth to the15th day. The results were as followings. 1. The body and cerebellum weights were more slowly increase of the the 2.5 Gy, 5 Gy and 10 Gy irradiation group compare to the control group, and the health condition of the 2.5 Gy group was a little bad. but the 10 Gy group was more severe and begun to die from the 12th day after birth. 2. The thickness, proliferation and migration of the 2.5, 5 and 10 Gy irradiated external granular cells from the maginal zone to the medullary area forming the molecular layer from the 6th day to the 15th day after birth were thinner, weaker and more slower according to the radiated dosages than the control group in the cresyl violet staining. 3. The proliteration, migration and lobulation of the 5 Gy radiated groups from the first day to the 15th day after birth were more weak, incomplete and irregular shape in the immunostaining with Dab, Cdk5, P35, calbindin and Zebrin antibody. 4. In the western blotting analysis using the Reelin, Dab, Cdk5 and P35 antibody. The Bands were in the 60 KD, 80 KD, 33 KD and 35 KD, and there were no differences between the control and irradiated groups in the molecular band except the Reelin. 5. As a results, the proliferation and migration of the outer granular and purkinje cells, and lobulation of the cerebellum by the several dosaege of the ${\gamma}$-ray radiation were proportionally incomplete according to dosage.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.679-685
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• 1998

We have studied, by a nonisotopic in situ DNA end-labeling (ISEL) technique, frequency of apoptosis in the external granular layer (EGL) of the cerebellum after whole-body irradiation of newborn mice and intestinal crypt cell of adult mice by gamma-rays from $^{60}Co$. The extent of changes following 2 Gy(10.9 Gy/min) was studied at 2, 4, 6, 8, 12, or 24h after exposure. The maximal frequency was found 4-8h after exposure. The mice that received 0.18, 0.36, 0.54, 1.08, 1.98, or 3.96 Gy were examined 6h after irradiation. Measurements performed after irradiation showed a dose-related increase in apoptotic cells in each of the mice studied. The dose-response curves were analyzed by a linear-quadratic model; frequency(%) of apoptotic cell in the newborn mice cerebellum was ($13.49{\pm}1.175$)D+$(-1.52{\pm}0.334)D^2$+0.048($r^2=0.981$, D = dose in Gy) and frequency(number per crypt) of apoptotic cell in the intestinal crypt of adult mice was ($3.857{\pm}0.420$)D+$(-0.535{\pm}0.120)D^2$+0.155($r^2=0.952$, D = dose in Gy). It provides the basis required for a better understanding of results which will be obtained in any further studies for biological responses of radiation using newborn and adult mice.