There are many modifications of eye shape and structure among fish, the general plan is similar throughout. This study was performed to comparative investigation for the lens shape and the interlocking pattern of lens fiber in genus Zacco (Z. temmincki and Z. platypus) and Pseudogobio (P. esocinus). The equatorial and axis diameter of lens for the classification of lens shape were measured by micrometer. And the interlocking patterns of lens fibers were observed by scanning electron microscopy (SEM). The lens shapes of Z. platypus and Z. temmincki were spherical (axis /equqtorial diameter = 1), but the lens shape of P. esocinus was subspherical type (axis / equqtorial diameter = 0.87). The interlocking patterns of lens fibers showed that Z. temmincki have an "anchor and socket" connection, Z. platy-pus have a "ball and socket" connection, and P. esocinus have a "rod and socket" connection. The results of this study may be utilized in the taxonomic keys for the classification of fish.
Axis, Cervical Vertebra ; Classification ; Microscopy, Electron, Scanning ; Platypus

Sonic hedgehog (Shh) has been known as an essential morphogen for the generation of motor neuron in developing spinal cord. However, motor neuron can be generated in Shh -/- ; Gli3 -/- or Gli2 -/- ; Gli3 -/- mutants although these mutants don't have Shh signaling. To find out the compensatory mechanism for the generation of motor neuron in Shh -/- ; Gli3 -/- mutant, we studied bone morphogenetic protein (BMP) antagonists including follistatin, flik and noggin, and retinoic acid signaling in this mutant. To study expressions of BMP antagonists, we performed in situ hybridization. To examine an activity of retinoic acid, we measured beta -galactosidase activity in retinoic acid response element (RARE) transgenic mouse. The expression of follistatin was reduced at both levels of forelimb and hindlimb in Shh -/- mutant compared to wild type embryo. It was restored at the level of forelimb but reduced at the level of hindlimb in Shh -/- ; Gli3 -/- mutant compared to wild type. The expression of flik was similar with wild type embryo at both levels of forelimb and hindlimb in Shh -/- mutant. The expression of flik was similar with wild type embryo at the level of forelimb however reduced in hindlimb level in Shh -/- ; Gli3 -/- mutant. The expression of noggin, a BMP antagonist, was increased in Shh -/- mutant. Activity of retinoic acid signaling was not affected in Shh -/- or Shh -/- ; Gli3 -/- mutants. From these results, we conclude that retinoic acid but not follistatin and flik, may be involved in the generation of motor neuron in Shh -/- ; Gli3 -/- mutant.
Animals ; Bone Morphogenetic Proteins ; Embryonic Structures ; Follistatin ; Forelimb ; Hedgehogs ; Hindlimb ; In Situ Hybridization ; Mice ; Mice, Transgenic ; Motor Neurons ; Response Elements ; Spinal Cord ; Tretinoin

The present study was designed to observe the expression patterns of NF-kappa B and AP-1, redox-sensitive transcription factors, and Bcl-2 and Bax, apoptosis repressing and promoting factors, respectively, upon repetitive cycles of short ischemia and reperfusion. Nine and thirty five weeks old Sprague-Dawley rats were subjected to the 3, 6, and 10 cycles of the ischemic process for 5 minutes followed by reperfusion for 5 minutes. The rats were divided by 5 groups, according to the time after treatment, such as 0, 3, 6, 24 and 72 hours. For short ischemia and reperfusion, left common iliac artery was occluded 3, 6, and 10 times for 5 minutes of ischemia followed by 5 minutes of reperfusion using rodent vascular clamps and left rectus femoris muscles were removed. The expression profiles and distribution of NF-kappa B, AP-1, Bcl-2, and Bax which were observed using immunohistochemical staining methods with 6 microgram thick paraffin sections of the rectus femoris tissue were as follows: The distribution of NF-kappa B was increased as the cycles of ischemia and reperfusion increased up to 3 hours after treatment. This phenomenon was prominent in 35 weeks-old rats. The distribution of AP-1 was increased as the cycles of ischemia and reperfusion increased up to 3 hours after treatment. This phenomenon was prominent in 9 weeks-old rats. The distribution of Bcl-2 was decreased as the cycles of ischemia and reperfusion increased up to 3 hours after treatment. The extent of such reduction was more prominent in 35 weeks-old rats than 9 weeks-old rats. The distribution of Bax was increased as the cycles of ischemia and reperfusion increased up to 3 hours after treatment. After 3 hours of treatment, Bax positivity was gradually decreased in 9 weeks-old rats, but increased in 35 weeks-old rats to reach a peak at 24 hour after reperfusion. The extent of enhancement in 9 weeks-old rats was higher than that in 35 weeks-old rats. In summary, multiple episodes of short ischemia and reperfusion altered the expression profiles of NF-kappa B, AP-1, Bcl-2, and Bax in the rectus femoris muscle at the similar extents in 9 and 35 weeks-old rats. Such alterations were more more increased when the episodes were more repeated.
Animals ; Apoptosis ; Iliac Artery ; Ischemia* ; Muscles* ; NF-kappa B* ; Paraffin ; Quadriceps Muscle* ; Rats* ; Rats, Sprague-Dawley ; Reperfusion* ; Rodentia ; Transcription Factor AP-1* ; Transcription Factors

Maternal alcohol abuse is thought to be the common cause of mental retardation. Even moderate maternal alcohol consumption may produce fetal alcohol effects with behavioral and learning difficulties, if the drinking is associated with malnutrition. Especially, continuous alcohol consumption during critical period of brain development is very likely to produce fetal alcohol effects. The aims of this study are to investigate whether exogenous thyroxine treatment to alcohol -fed dams may ameliorate the detrimental effects of alcohol on the postnatal development of BDNF -containing Purkinje cell of the cerebellar cortex of the offspring. The morphological features of the growth and maturation were observed at 0, 7, 14, 21, 28 postnatal days via immunohistochemistry. In addition, electron microscopic finding of BDNF -containing Purkinje cell at P14 was also examined. Time -pregnant rats were divided into three groups. Alcohol -fed group received 35 calories of liquid alcohol diet daily from gestation day 6; control pair -fed group was fed a liquid diet in which dextrin replaced alcohol isocalorically; alcohol +/-T4 group received 35 calories liquid alcohol diet and exogenous thyroxine subcutaneously. As a result, a similar developmental pattern of BDNF -immunoreactive Purkinje cells was observed in control pair - fed and alcohol+/-T4 group on and after P14. These cells of alcohol -fed group showed immature features. Single -layer arrangement of these cells in alcohol -fed group was not completely achieved throughout postnatal life. Electron microscopic observations of BDNF -immunoreactive Purkinje cells at P14 revealed large nucleus, small cytoplasm, small amount of ribosomal collection and rudimentary cytoplasmic organelles in alcohol -fed group. The morphology of BDNF -immunoreactive Purkinje cell in alcohol +/-T4 group was similar to that in control pair -fed group. It was characterized by numerous short segments of rough endoplasmic reticulum, many of which showed a tendency of parallel alignment that suggested an attempt at Nissl body configuration. The cytology of Golgi complexes was also found within the cytoplasm in perinuclear location. Those observed differences of postnatal maturation patterns between alcohol -fed and alcohol +/-T4 group may indicate the beneficial effects on the postnatal development of BDNF -containing Purkinje cells in cerebellar cortex in the pups of thyroxine -treated alcohol -exposed dams. These results suggest that the increase of BDNF synthesis during early postnatal life caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effects as a result of the dysthyroid state following maternal alcohol abuse.
Alcohol Drinking ; Alcoholism ; Animals ; Brain ; Brain-Derived Neurotrophic Factor ; Cerebellar Cortex ; Cerebellum* ; Critical Period (Psychology) ; Cytoplasm ; Diet ; Drinking ; Endoplasmic Reticulum, Rough ; Golgi Apparatus ; Immunohistochemistry ; Intellectual Disability ; Learning ; Malnutrition ; Organelles ; Pregnancy ; Purkinje Cells ; Rats* ; Thyroxine*

We have examined the factors related to hair growth regulation during postnatal growth periods in C57BL/6N, hairless and alopecia areata mice model. We first studied the number, localization and granulation status of skin mast cells during postnatal growth periods by toluidine blue, alcian blue and H& E staining methods. We second studied immunoreactive density of neuropeptide (substance P, CGRP, CRF, CRF -receptor, CRF -binding protein) in skin by immunohistochemical methods. We third studied changes of subpopulation of splenocytes and thymocytes during postnatal growth periods in C57BL/6N, hairless and alopecia areata mice model by flow cytometry. The results were as follows : In C57BL/6N mice, the number of mast cells was decreased from 1day to 35 day during postnatal growth period. But skin of alopecia areata model mice in 21 day, the number of mast cells was more increased than that of normal skin. Immunoreactive density of neuropeptide (Substance P, CGRP, CRF, CRF -receptor, CRF -binding Protein) in skin during postnatal growth periods in C57BL/6N mice was weakly stained in 1, 3, 8 day, but immunoreactive density of neuropeptide was heavily stained in 35day and 21 day (no pigment). Splenic B and T lymphocytes were gradually increased with growth, and significantly increased in 29 days -old (hairless, pigment group) than 21 days -old (hairless, no pigment group) C57BL/6N mice. Especially, CD4 positive Th cells in splenic T lymphocytes were markedly increased. But thymic T lymphocytes were not shown any differences. In hairless mice, the number of mast cells was not changed from 3 day to 7 day, but more increased from 13day to 21day. Immunoreactive density of neuropeptide (substance P, CGRP, CRF) in skin during postnatal growth periods in hairless mice was weakly stained in 3, 7 and 13 day. but immunoreactive density of neuropeptide was heavily stained from 17 to 56 day. Splenic B and T lymphocytes were most highly increased in 13 days -old genetically hairless mice. And CD4 positive Th cells and CD8 positive Tc cells in splenic T lymphocytes were significantly increased. And also thymic Th lymphocytes were increased in 13 days -old and 17 days -old hairless mice. These experiment suggest that the factors related to hair loss were mast cells, immunoreactive density of neuropeptide (substance P, CGRP, CRF) and immune response during postnatal growth periods in C57BL/6N, hairless and alopecia areata mice model.
Alcian Blue ; Alopecia Areata* ; Alopecia* ; Animals ; Flow Cytometry ; Hair* ; Lymphocytes ; Mast Cells ; Mice* ; Mice, Hairless ; Neuropeptides ; Skin ; Substance P ; T-Lymphocytes ; Thymocytes ; Tolonium Chloride

Location of the modiolous and morphological variations of the risorius and zygomaticus major muscles are related to the facial expression. The zygomaticus major, levator labii superioris, depressor labii inferioris, depressor anguli oris, risorius, orbicularis oris, buccinator and levator anguli oris muscles insert on the lateral border of the lip, forming the modiolus and mutually associating each other for functioning. The knowledge of the location of the modiolus and surrounding structures are essential to anatomy, prosthodontics, linguistic, physiology and computer simulation based on facial expressions. The authors examined the location of the modiolus, the morphological variations and anatomical relationship of risorius and zygomaticus major muscle to understand the features of the smile of Korean by dissecting 39 cadavers. The location of the modiolus can be showed as three types, according to their height related to the intercheilion horizontal line. Type A that modiolus locate at the intercheilion line was shown in 20 sides (26.0%), type B that modiolus locate above the intercheilion line was shown in 12 sides (15.6%), then type C that modiolus locate under the intercheilion line was shown in 45 sides (58.4%). Most modioli located at 10 ~20 mm lateral to the mouth corner and 0 ~10 mm below the intercheilion line. The risorius muscle was classified into five types by directions of muscle fibers. The depressor anguli oris -risorius type (type I) was observed in 31 sides (40.2%), the platisma -risorius type (type II) was observed in 30 sides (39.0%). Previously, it has been known that zygomaticus major muscle attaches to the modiolus mainly as one bundle. However, the results were clearly shown that two bundles of the zygomaticus major muscle attaches to the modiolus and the position of the mouth edge in 18 sides (23.4%). To sum it up, facial expression is of fundamental importance concerning the morphological variations and these results also can be considered for the facial reconstruction surgery and computer animation department.
Cadaver ; Computer Simulation ; Facial Expression* ; Linguistics ; Lip ; Mouth ; Muscles ; Physiology ; Prosthodontics

Eruptive movement of the tooth germ accompanies the resorption and formation of the surrounding alveolar bone. Bisphosphonate has effects on osteoclasts to inhibit bone resorption, being currently used for the treatment of bone disease such as osteoporosis. This study was conducted to elucidate effects of bisphosphonate on tooth eruption in growing rats. Alendronate, a derivative of bisphosphonate, was injected into newborn rats at the concentration of 0.5 mg/kg and 2.5 mg/kg for 3, 7 and 10 days. The incisor and molar teeth normally erupted at postnatal 22 days in the saline-treated control group, whereas no teeth could not be seen in both groups of the alendronate treatment. No conspicuous histological changes could not be found in the teeth germs in the alendronate treated rats. The number of osteoclasts in both the first and the second molars was maximum on the postnatal day 10. The number of osteoclasts significantly increased by both 3 and 7 days of alendronate treatment (p< 0.01). But the number significantly decreased by 10 days of alendronate treatment in both concentrations (p< 0.01). The size of osteoclasts was not different between the control and alendronate groups. Thus, these results suggest that bisphosphonate acts on osteoclast formation to inhibit bone resorption and subsequent tooth eruption.
Alendronate ; Animals ; Bone Development* ; Bone Diseases ; Bone Resorption ; Humans ; Incisor ; Infant, Newborn ; Molar ; Osteoclasts ; Osteoporosis ; Rats* ; Tooth Eruption ; Tooth Germ ; Tooth*

This study is aimed to investigate the signal transduction pathway of amyloid beta peptide (Abeta)-induced neuronal toxicity and the inhibitory effects of epigallocatechin gallate (EGCG), one of the major constituents of green-tea and the potent anti-oxidant, on the nerve cell damage in PC12 cells. Cellular toxicity was estimated by MTT assay and observation of morphological changes in PC12 cells. By using the methods such as measurement of Reactive Oxygen Species (ROS), western blot and RT-PCR, the underlying mechanisms and signal transduction pathway of Abeta- induced neurotoxicity and the inhibitory effects of EGCG were examined. Abeta-induced cellular toxicity was found in a dose dependent manner. This is confirmed by morphological observations of cultured cells such as findings of cell death similar to apoptosis. Abeta-induced neurotoxicity was effectively inhibited by EGCG pretreatment. Moreover, EGCG reduced ROS as same potent as he NAC (N-acetyl cystein), the ROS scavenger. Among the several process of signal transduction for cell death, a intracytoplasmic cytochrome c, the protein associated with the mitochondria- dependent pathway, was increased from 12 hours after Abeta treatment and the increased cytochrome c by Abeta was blocked by EGCG. Expression levels of Bax/Bcl-2 in relation to intracytoplasmic release of cytochrome c were examined by RT-PCR. Abeta up-regulated Bax expression but did not affect Bcl-2 expression. EGCG was found to block the effect of Abeta-induced Bax increase. From these results, it is speculated that Abeta-induced neuronal toxicity may be assumed to be affected by ROS and the mitochondria-dependent pathway of cell death as well. EGCG, besides having the role of anti-oxidant, is found to have a protective effect against Abeta-induced neurotoxicity through the inhibition of the expression of the protein associated with the mitochondria-dependent cell death pathway.
Amyloid beta-Peptides ; Amyloid* ; Animals ; Apoptosis ; Blotting, Western ; Cell Death ; Cells, Cultured ; Cytochromes c ; Neurons* ; PC12 Cells ; Reactive Oxygen Species ; Signal Transduction

Astragalus membranaceus is used as a natural herbal medicine in East Asia for preventing carcinogenesis and reducing side effects induced by chemotherapy in cancer patients. Although the mechanism of anti-tumor activity is not known, the polysaccharides may potentiate the host defense mechanism through the activation of immune system. The objective of this study is to investigate the mechanism by which APS activates macrophages. To analyze macrophage activation and iNOS gene expression, we performed nitrite generation assay, immunohistochemistry, and RT-PCR. In the present study we show that a polysaccharide isolated from the Astragalus membranaceus (Astragalus Polysaccharide, APS) significantly induces nitric oxide (NO). Immunohistochemical staining of inducible NO synthase (iNOS) showed that the increase of NO was due to the induction of iNOS production. To further study the mechanism responsible for the induction of iNOS, we investigated the effect of APS on the iNOS mRNA expression. RT-PCR analysis showed that APS produced significant induction of iNOS gene expression. In conclusion, we demonstrate that a polysaccharide isolated from Astragalus membranaceus stimulates macrophages to generate NO through the activation of iNOS gene expression.
Astragalus membranaceus* ; Carcinogenesis ; Drug Therapy ; Far East ; Gene Expression ; Herbal Medicine ; Humans ; Immune System ; Immunohistochemistry ; Macrophage Activation ; Macrophages ; Nitric Oxide ; Nitric Oxide Synthase ; Polysaccharides ; RNA, Messenger

In skeletal muscles, oxygen free radicals generated during ischemia -reperfusion are known as inducers that cause cellular injury and apoptosis and contribute to the pathogenensis of reperfusion injury. Ischemia -reperfusion for 2 hours may cause reversible changes, while prolonged ischemia -reperfusion causes irreversible changes. Following ischemia -reperfusion, diverse signals are transduced to induce a variety of gene expression. Ischemic preconditioning, defined as brief episodes of ischemia and reperfusion, is known to provide protection from the consequences of prolonged ischemia followed by reperfusion. NF -kappa B is a transcription factor that activated during ischemic preconditioning and ischemia -reperfusion. It initiates inflammation through inducing transcription of proinflammatory, procoagulant and vasoactive gene, while mediates the expression of cytoprotective proteins that block apoptosis or inhibit inflammation. The present study was performed to study the change of NF -kappa B immunoreacitvity in rat anterior tibialis and soleus muscles in response to ischemia -reperfusion and preconditioning. Experimental animals, Sprague -Dawley rats (250 ~300 g), were divided into 6 groups; 1) control, 2) ischemic preconditioning, 3) 2 hours of ischemia, 4) 4 hours of ischemia, 5) 2 hours of ischemia after ischemic preconditioning, 6) 4 hours ischemia after ischemic preconditioning. For ischemic preconditioning, left common iliac artery was occluded three times for 5 minutes followed by 5 minutes of reperfusion using vascular clamp. Ischemia was done by occlusion of the same artery for 2 or 4 hours. The specimens of tibialis anterior and soleus muscles were obtained 0, 1, 3, 6, and 24 hours after onset of reperfusion. The specimens were paraffin sectioned at 6 micrometer and NF -kappa B expression was examined using immunohistochemical methods. The results obtained were as follows : 1. In normal control group, immunoreactivity of NF -kappa B was moderate to strong in tibialis anterior muscles and weak in soleus muscles. 2. In tibialis anterior, immunoreactivity of NF -kappa B was decreased in 2 and 4 hours of ischemia comparede with normal control group. In soleus muscle, immunoreactivity of NF -kappa B was decreased in 2 hours of ischemia but it was comparable to that of normal control group in 4 hours of ischemia. 3. Ischemia for 4 hours induced more remarkable change in NF -kappa B immunoreactivity than that for 2 hours. 4. After ischemic preconditioning, changes in NF -kappa B immunoreactivity after 2 and 4 hours of ischemia were decreased compared with normal control group. 5. In ischemia for 2 and 4 hours, changes in NF -kappa B immunoreactivity of tibialis anterior muscles were more severe than that of soleus muscles. These results suggest that in the skeletal muscle, changes in NF -kappa B immunoreactivity of 4 hours of ischemia were more remarkable than that of 2 hours ischemia, and changes in NF -kappa B of tibialis anterior muscles were more remarkable than that of soleus muscles. Ischemic preconditiong attenuated the alteration of the NF -kappa B immunoreactivity induced by ischemia -reperfusion in the muscles.
Animals ; Apoptosis ; Arteries ; Free Radicals ; Gene Expression ; Iliac Artery ; Inflammation ; Ischemia ; Ischemic Preconditioning ; Muscle, Skeletal ; Muscles* ; Oxygen ; Paraffin ; Rats* ; Reperfusion ; Reperfusion Injury ; Transcription Factors