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Morphometric Study of the Testicular Interstitium of the Rat during Postnatal Development.

Korean Journal of Anatomy.1999;32(6):849-858.

The purpose of this morphometric study was to obtain quantitative information on the rat testis interstitium during postnatal development. Eight groups of male rats aged 1, 7, 14, 21, 28, 40, 60 and 90 days (n=5 rats per group) after birth were used. Tissue from perfusion-fixed testes was embedded in Epon-Araldite; and sections were subjected to morphometric measurements at the light microscopic level, using point counting method for volume densities and the Disector technique for numerical densities (the number of cells per unit volume of testis). The volume density of the interstitium represents 66% of the testicular parenchyma at day 1. This proportion progressively diminishes during development to reach a value of 8% at day 90. The absolute volume of blood vessels, macrophages, and endothelial cells increased with age. The absolute volume of lymphatic spaces, pericytes and myoid cells were greater at 90 days than at any other age. The absolute volume of fetal Leydig cells per testis was unchanged from 1 (0.07 mm3) to 14 (0.1 mm3) days, despite a decrease in the volume density. The number of mesenchymal cells, myoid cells, macrophages, endothelial cells, and pericytes per testis increased with age. The number of fetal Leydig cells per testis did not change from days 1 (0.054 million)~21 (0.070 million) although on day 21 (615 micrometer3) an average fetal Leydig cells was smaller in volume than at earlier ages (days 1 (1338 micrometer3)~14 (1296 micrometer3)). Adult Leydig cells were recognized at day 14 and their absolute volume and number per testis increased from 14 (0.5 mm3, 0.6 million) to 90 (52.83 mm3, 21.14 million) days. The average volume of a adult Leydig cell increased significantly with age and reached maximum size by 60 days (2548 micrometer3) of age where the volume is nearly three times bigger than that of at day 14 (832 micrometer3). No change in the average volume of the macrophages could be detected in this study groups. The average volume of the mesenchymal cells decreased significantly from day 1 (812 micrometer3) to day 14 (385 micrometer3) then increased until day 28 (901 micrometer3) at which the volume is maximum and declined significantly thereafter.
Adult ; Animals ; Blood Vessels ; Endothelial Cells ; Humans ; Leydig Cells ; Macrophages ; Male ; Parturition ; Pericytes ; Rats* ; Testis

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Maturation of the Neurokinin 1 (NK1) Receptor Immunoreactive Amacrine Cells in the Rat Retina during Postnatal Development.

Tae Hoon KANG ; Jin Woong CHUNG

Korean Journal of Anatomy.2003;36(5):435-441.

We examined the morphological maturation of amacrine cells expressing neurokinin 1 (NK1) receptor, whose ligand is substance P, in the rat retina, focusing on the period from postnatal day 5 (P5) when the outer plexiform layer is formed, to postnatal day 13 (P13) when the eyes open, with immunohistochemistry using a specific antiserum against NK1 receptor, and we compared maturing NK1 receptor-immunoreactive (NK1 receptor-IR) amacrine cells with adult one. In the adult retina, numerous NK1 receptor-IR amacrine cells were located in the inner part of the inner nuclear layer (INL) adjacent to the inner plexiform layer (IPL), and their processes emerging from the somata branched and stratified at 1, 2, and 5 strata of within the IPL. NK1 receptor-IR amacrine cells were already observed at P5. The cell bodies were located in the inner INL away from the IPL and their processes branched and formed two distinct bands in the IPL. Afterwards, somata of NK1 receptor-IR amacrine cells moved toward the inner part of the INL, and thus, were located in the INL adjacent to the IPL. Their processes formed three distinct bands at P10 and then, at P13, three bands occupied the same strata as those of the adult, which were posed at 1, 2, and 5 strata of the IPL. During the postnatal development, most of NK1 receptor-IR amacrine cells directly extended one or a few primary dendrites toward the IPL and formed the strata. However, some of the labeled cells located at the outermost row had horizontal processes emerging from their primary dendrites, and these horizontal processes branched and formed plexuses in the INL. The NK1 receptor-IR amacrine cells with horizontal processes were frequently observed at P7, rarely at P10, and not at P13 and in the adult. These results indicate that the NK1 receptor-IR amacrine cells of the rat retina morphologically mature by way of migration of their somata within the INL and formation of distinct processes during postnatal development, and suggest that they morphologically and functionally complete the maturation process about the time of P13.
Adult ; Amacrine Cells* ; Animals ; Dendrites ; Humans ; Immunohistochemistry ; Rats* ; Retina* ; Substance P

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Double Immunohistochemical Study on the Postnatal Development of Somatostatin - and Neuropeptide Y - Immunoreactive Neurons in Rat Cerebral Cortex.

Eun Young LEE ; Tae Soo LEE ; Sang Ho BAEK ; Jung Ik CHA

Korean Journal of Anatomy.1997;30(5):483-494.

The postnatal development of somatostatin [SOM]- and neuropeptide Y[NPY]- immunoreactive[ir] neurons were examined in rat cerebral cortex considering their coexistence in cortical neurons. Using double immunohistochemical staining for SOM and NPY with diaminobenzidine and benzidine dihydrochloride as chromogens, we subdivided immunoreactive cells into double-labeled SOM/NPY-, SOM only-, and NPY only-ir neurons. Interestingly, SOM/NPY- and SOM only-ir neurons were detectable even at the day of birth, in contrast to NPY only-ir cells which first appeared in most cortices from two weeks of age. The morphological features of double-labeled SOM/NPY neurons were not identical to those SOM only- and NPY only-ir neurons. No apparent changes in the shape and size of single-labeled neurone occurred with age ; throughout their postnatal life they were round and ovoid, had a thin rim of perinuclear cytoplasm, and short processes. In contrast, the features of SOM/NPY-ir neurons were not consistent during postnatal life. By day P7, these neurons showed immature features ; they began to show more advanced neuronal characteristics by week P2, when they had a larger and more intensely-stained cytoplasm. In addition, their processes were longer, thicker and more complex than at earlier ages. At this age, SOM/NPY-ir somata were close to their maximum size. From week P4, they became smaller and were lightly labeled. SOM/NPY-ir somata were larger than SOM only- and NPY only-ir somata at and after two weeks of age. The present results showing different postnatal maturation patterns such as time of appearance and morphological features suggest that double-labeled SOM/NPY and single-labeled neurons might be different populations regulated by different mechanisms in their development, and with different functional properties during development.
Animals ; Cerebral Cortex* ; Cytoplasm ; Neurons* ; Neuropeptide Y* ; Neuropeptides* ; Parturition ; Rats* ; Somatostatin*

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Proliferation of Mature T Cells in the Periphery of the Mice.

Dong Sup LEE ; Wang Jae LEE

Korean Journal of Anatomy.1999;32(5):717-726.

In order to find out the proliferative ability and functional capacity of peripherally expanded mature T cells, syngeneic lymph node cells were injected into thymectomized and irradiated adult mice. Donor and host T cells were evaluated in specific time points. The results were as followings. 1. As compared to the engrafted lymphocytes after transfer, we observed 54~60 fold increase in CD4 T cells and 113~130 fold increase in CD8 cells, that is, CD8 cells proliferated more than CD4 cells in the periphery. 2. Mature T cells proliferated in the periphery got activated/memory phenotype in the early period of expansion but they upregulated CD45RB and CD62L in the later period. CD44 level remained high. 3. T cells maintained their response against soluble antigen, KLH but the reactivity against some polyclonal T cell activators, such as anti-CD3 antibody and concanavalin A were reduced severely in the 6 month expanded group. 4. The leftover host T cells also proliferated in the space and they behave similar to donor T cells in proliferation and change of activation/memory markers.
Adult ; Animals ; Concanavalin A ; Humans ; Lymph Nodes ; Lymphocytes ; Mice* ; Phenotype ; T-Lymphocytes* ; Tissue Donors

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In vivo Study on the Japanese Encephalitis: Viral Localization and Histopathology in the Mouse Brain.

In Beom KIM ; Soo Lim CHAE ; Woo Young CHOI ; Chan PARK ; Young Ran JOO ; Hae Wol CHO ; Keun Yong PARK

Korean Journal of Anatomy.2003;36(5):427-433.

Japanese encephalitis is a potentially lethal disease of the central nervous system caused by infection with Japanese encephalitis virus (JEV). JEV is the most common cause of encephalitis over a large part of eastern Asia. To establish and characterize in vivo model to study the Japanese encephalitis, the immunohistochemical localization of JEV and the histopathological finding were investigated in the brains of young adult mice infected with JEV by intraperitoneal inoculation. JEV was localized to neurons in discrete regions of the brain. Histopathological finding showed typical pattern of acute viral encephalitis, such as inflammatory cell infiltration in brain parenchyme and perivascular cuffs of mononuclear cells. These results suggest that this in vivo system can be used to study the mechanism of virus entry into the brain, cell specific tropism, and pathophysiology in Japanese encephalitis.
Animals ; Asian Continental Ancestry Group* ; Brain* ; Central Nervous System ; Encephalitis ; Encephalitis Virus, Japanese ; Encephalitis, Japanese* ; Encephalitis, Viral ; Far East ; Humans ; Immunohistochemistry ; Mice* ; Neurons ; Tropism ; Virus Internalization ; Young Adult

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Expression of the TGF-beta Family during the Development of Rat Eye.

Ki Young KANG ; Jin Ghi BAEK ; Hyun KIM

Korean Journal of Anatomy.2003;36(5):417-426.

The development of eye is a very dynamic process in which various cells and tissues from diverse originneuroectoderm, surface ectoderm and mesenchyme, undergo induction, mitosis and differentiation. During this process advances, many growth factors are involved. Transforming growth factor betas (TGF-betas) are the pivotal growth factors during cellular growth, differentiation, matrix formation, migration and wound healing. Especially during the development of tissues, TGF-beta s are secreted and induce growth and differentiation of adjacent cells. To investigate the functions of TGF-betas during the development of eye, we studied the expression of TGF-betas in rat embryos. For this study, Sprague-Dawley rat embryos were taken on the 11th 13th, 15th, 17th and 19th day of gestation. To confirm the morphological changes of eyes on each developmental stage, sections were stained with hematoxylin and eosin. Then, to document the expressive patterns of TGF-beta1, beta2, beta3 and TGF-beta receptor type II (TbetaR), immunohistochemistry was applied. The results were obtained as follows: TGF-beta1 was partially expressed in the lens only on the 19th day of gestation. TGF-beta2 was expressed in the retina and lens on the 11th and 13th day, while it was detected in most areas of the eye from the 15th to 19th day. TGF-beta3 was expressed only in the pigment cell layer of the retina on the 11th day, but it was detected in various areas along the advance of the development. Immunoreactivity of TGF-beta3 was always weaker than TGF-beta2. TbetaR showed strong immunoreactivity in where the reactivity of TGF-beta2 was detected. These results indicate that TGF-beta2 and beta3 play important roles during the development of eye.
Animals ; Cornea ; Ectoderm ; Embryonic Structures ; Eosine Yellowish-(YS) ; Hematoxylin ; Humans ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; Mesoderm ; Mitosis ; Pregnancy ; Rats* ; Rats, Sprague-Dawley ; Receptors, Transforming Growth Factor beta ; Retina ; Transforming Growth Factor beta* ; Transforming Growth Factor beta1 ; Transforming Growth Factor beta2 ; Transforming Growth Factor beta3 ; Wound Healing

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Ultrastructure of Fronto-Accumbens Synapses after Prefrontal Decortication of the Rat.

Hyoung Gon SONG ; E Tay AHN ; Jeong Sik KO ; Kyung Ho PARK

Korean Journal of Anatomy.2003;36(5):405-416.

Prefrontal cortex is called psycological cortex, since it deals with making up of individual personality, regulation of personal depth of feeling, working memory, planning, maintaining attention, etc. Whereas, nucleus accumbens (septi) is called the center of reward and motivation or the center of pleasure, since it deals with feeding, drinking, sex, exploration, appetitive learning, drug addiction, etc. Present study was aimed at the proving the prefronto-accumbens input ultrastructurally. Sprague Dawley rats anesthetized with sodium pentobarbital and were removed their prefrontal cortex with suction instrument. Two days following the operation, heads of rats were fixed by perfusion of with 1% glutaraldehyde-1% paraformaldehyde solution via left ventricle. Peristaltic pump was used during perfusion. Two hours later, brains were removed and refixed for 24 hours in the refrigerator, and small tissues of the nucleus accumbens were punched out with punching needle. Tissue blocks were fixed in 2% osmic acid for 2 hours and were embedded in araldite mixture. Ultrathin sections stained with uranyl acetate-lead citrate solution were observed with JEOL 100 CX II electron microscope. In the nucleus accumbens, some axodendritic terminals and axospinous terminals were found degenerated, and volume of activated glial cytoplasm was increased. The degenerated terminals were seen isolated from intact structures by activated glial processes and removed by glial cytoplasm. The result confirms that axon terminals coming from prefrontal cortex input to the spiny neurons of nucleus accumbens septi, on their dendrites and/or dendritic spines.
Animals ; Brain ; Citric Acid ; Cytoplasm ; Dendrites ; Dendritic Spines ; Drinking ; Head ; Heart Ventricles ; Humans ; Learning ; Memory, Short-Term ; Motivation ; Needles ; Neurons ; Nucleus Accumbens ; Osmium Tetroxide ; Pentobarbital ; Perfusion ; Pleasure ; Prefrontal Cortex ; Presynaptic Terminals ; Rats* ; Rats, Sprague-Dawley ; Reward ; Sodium ; Substance-Related Disorders ; Suction ; Synapses*

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Reduction of Spinal Cyclooxygenase-2 with Transcutaneous Electrical Nerve Stimulation and Cold Therapy in Rats of Carrageenan-induced Inflammatory Muscle Pain.

Yun Woong PAEK ; Yun Won CHAE ; Jung Woo KIM

Korean Journal of Anatomy.2003;36(5):397-403.

The purpose of this study was to investigate the effects of transcutaneous electrical nerve stimulation (TENS) and cold therapy on cyclooxygenase-2 (COX-2) expression in lumbar spinal cord and on secondary hyperalgesia produced by muscle pain. Muscle pain was experimentally induced by injection of 2% carrageenan into gastrocnemius muscle of Sprague-Dawley rats. For assessment of secondary hyperalgesia, the rats were tested for paw withdrawal latency (PWL) and tail flick latency (TFL) to heat stimulus. COX-2 expression in lumbar cord was examined using RT-PCR and immunohistochemistry. Secondary hyperalgesia to heat stimulus was significantly reduced in the both TENS- and cold-treatment groups, compared to the control group. The COX-2 mRNA levels were down-regulated in the lumbar spinal cord in the both TENS- and cold-treatment groups. In addition, COX-2 immunoreactivity was decreased in the dorsal horn of the lumbar spinal cord in the both TENS- and cold-treatment groups. These results suggested that decreased COX-2 expression in the lumbar spinal cord of the subjects receiving TENS treatment and cold therapy might be an important factor for reducing secondary hyperalgesia produced by muscle pain.
Animals ; Carrageenan ; Cryotherapy* ; Cyclooxygenase 2* ; Horns ; Hot Temperature ; Hyperalgesia ; Immunohistochemistry ; Muscle, Skeletal ; Myalgia* ; Rats* ; Rats, Sprague-Dawley ; RNA, Messenger ; Spinal Cord ; Transcutaneous Electric Nerve Stimulation*

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The Effect of Tumor Necrosis Factor-Alpha to Glomerular Epithelial Cells in Glomerular Permeability.

Cheol Woo KO ; Kyung Hee LEE ; Min Hyun CHO ; Dong Kyu CHO ; Jae Tae LEE ; Hee Jung CHO

Korean Journal of Anatomy.2003;36(5):389-395.

Minimal Change Disease (MCD) is the most common primary nephrotic syndrome in children. Some suggested that tumor necrosis factor-alpha (TNF-alpha) are involved in the pathogenesis of MCD. This study was done to see changes of plasma and urinary TNF-alpha, and its effect on determination of permeability of glomerular basement membrane (BM) contributed by heparan sulfate proteoglycan (HSPG). Study patients consisted of 19 biopsy-proven MCD children aged 2-15 years old. Both plasma and urinary TNF-alpha were measured. Employing the Millicell system, TNF-alpha was screened for the permeability factors. We examined whether TNF-alpha regulated BM HSPG gene expression and HS synthesis in the glomerular epithelial cells (GECs). Urinary TNF-alpha during relapse was also significantly increased (364.4+/-51.2 vs 155.3+/-20.8, 36.0+/-4.5 ng/mg.cr) (p<0.05). However, the negative results were obtained in the permeability assay using the Millicell system. No difference was seen in BM HSPG gene expression and HS synthesis in the GECs. Therefore, it seems that TNF-alpha may not play a disease-specific role in the pathogenesis of MCD.
Child ; Epithelial Cells* ; Gene Expression ; Glomerular Basement Membrane ; Heparan Sulfate Proteoglycans ; Humans ; Nephrosis, Lipoid ; Nephrotic Syndrome ; Permeability* ; Plasma ; Recurrence ; Tumor Necrosis Factor-alpha*

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The Efficient Gene Delivery into Human Mesenchymal Stem Cells Using Retroviral Vectors.

Sung Soo KIM ; Bum Jun KIM ; Haeyoung Suh KIM

Korean Journal of Anatomy.2003;36(5):381-387.

Human mesenchymal stem cells (hMSCs) are multipotent stem cells that can differentiate into several mesenchymal lineage cells. In this study, we established an efficient method for gene delivery into these cells. Non-viral transfection reagents that were commercially available yielded 5% efficiency. In contrast, a retroviral vector yielded more than 46% transduction, which was further increased to 90% by repetitive infection. Retroviral transduction did not alter the multipotency of hMSCs. Thus, the cells retained the potential to differentiate into adipogenic, chondrogenic, or osteogenic lineages. The conditions established in this study will contribute to development of trans-differentiation methods of hMSCs into non-mesodermal lineage cells and thereby facilitate their possible use as vehicles for autologous transplantation in both cell and gene therapy for various diseases.
Autografts ; Genetic Therapy ; Humans* ; Indicators and Reagents ; Mesenchymal Stromal Cells* ; Multipotent Stem Cells ; Retroviridae ; Transfection ; Transplantation, Autologous ; Zidovudine*

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