• Applied Microscopy
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• v.25 no.1
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• pp.65-74
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• 1995

An immunocytochemical study of GABA-positive neuronal elements was performed at the electron microscopic level to examine subcellular distribution of the inhibitory neurotransmitter in the dog basilar pons. Electron-dense reaction product was observed in neuronal somata and dendritic processes. One or more unlabeled axon terminals made asymmetric synaptic contacts with these GABAergic somatic and dendritic profiles. A large number of GABA-positive axon terminals were also observed. They made symmetric as well as asymmetric synaptic contacts with unlabeled dendritic profiles. In axo-axonic synapses, postsynaptic axon-like processes were consistently GABA-immunoreactive. These observations suggest that the inhibitory local circuit neurons in the dog basilar pons play a major role in cerebro-ponto-cerebellar circuitry by integrating various afferent inputs and conveying them into the cerebellar cortex and the deep cerebellar nuclei.

• Animal cells and systems
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• v.5 no.3
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• pp.263-266
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• 2001

Metallothionein (MT) is induced in the regenerating rat liver. We have investigated expression of MT gene by RT PCR as well as specific localization of MT by immunocytochemistry in regenerating rat liver after partial hepatectomy (PH). MT mRNA level started to increase from 1 h and reached the peak at 8 h after PH. The level decreased gradually by 24 h, and became similar to that of control group. In the immunocytochemical study, in all groups treated with primary antibody, immunogold particles indicating the presence of MT were evenly distributed throughout both cytoplasm and nucleus of the rat hepatocytes. Within the nucleus, the gold particles appeared to be intensely localized in the areas of euchromatin and nucleolus. Within the cytoplasm, gold particles did not seem to adhere to mitochondria or Iysosomes, but were freely distributed. However, rough endoplasmic reticulum was the obvious compartment on which the gold particles were localized. Time course of MT immunoreactivity revealed that distribution of gold particles in hepatocytes increased gradually by 24 h, and decreased at 48 h after PH. Briefly, PH resulted in the sharpest increase in the expression of MT mRNA at 8 h and in the immunoreactivity of MT at 24 h, respectively. It is suggested that the increase of MT mRNA expression, the intensity of immunoreactivity and the specific localization of MT may be associated with the compensatory cell proliferation followed by PH.

• The Korean Journal of Zoology
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• v.35 no.4
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• pp.448-455
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• 1992

생쥐에 hCG를 투여한 후 자궁벽에서 hCG와 결합하는 세포의 종류를 판별하고, 그 세포가 hCG에 의해 어떠한 영향을 받는지를 알아보기 위하of 면역세포화학적 염색을 수행하였다. p-hCG에 대한 단일크론항체에 양성반응을 나타내는 세포는 주로 자궁벽의 자궁근막 삼각과 자궁내막 기저층에 분포하였으며, 세포질에서 양성반응을 일으킨 것으미 관찰되었다. hOG를 투여하고 3, 6, 24시간 후에 각각 자궁벽라 자궁근막에서 양성반응세포를 관찰한 결과, 양성반응세포의 이동현상을 관찰할 수 있었다. 즉, 자궁근막에서 보면, 3시간 후에는 자궁근막의 혈관내부와 주변에서 다수가 관찰되었으며, 6시간 후에는 소수가 관찰되었고, 24시간 후에는 관찰되지 않았다. 또한 동일 시간에 적출된 자궁벽에서는 6시간에서 가장 많은 수의 양성세포가 관찰되었다. 따라서 자궁벽과 자궁근막에서 관찰되는 양성반응세포의 수 사이에는 반비례적 상관관계를 나타냈다. 한편, 난소를 적출한 생쥐의 자궁에서는 양성반응세포의 수가 현저히 감소된 것으로 나타났으며, hematoxvlin-eosln 염색 결과 양성반응세포로 추정되는 세포의 세포질에서 호산성과림이 관찰되었다. 이와 같은 결과로 볼 때, 양성세포는 GMG(granulated metrial gland) 세포로 판단되었으며, 이 세포의 이동은 스테로이드호르몬에 의해 영향을 받는 것으로 사료되었다.

• Animal cells and systems
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• v.16 no.2
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• pp.114-120
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• 2012

In echinoderms, the SALMFamide neuropeptides sharing the SxL/FxFamide motif seem widespread throughout the phylum and may be important signalling molecules that mediate various physiological functions. Recent identification of S1 and its analogues, MagS3 and MagS4, along with the S2 analogue, MagS2 from the starfish $Marthasterias$ $glacialis$, indicated that SALMFamides in the class Asteroidea are more diverse than previously thought. Further, isolation of the neuropeptides from the radial nerve cord and studies on pharmacological actions of the neuropeptides on the cardiac stomach warrant studies on the tissue distributions of these peptides in both the nervous and digestive systems. In the present study, antisera raised against an S1 analogue, KYSALMFamide, and an S2 analogue, KYSGLTFamide, were used to localize the distribution patterns of the S1- and S2-like immunoreactivities (S1-IR/S2-IR) in the nervous and digestive systems of the starfish. In the nervous system, cell bodies in the ectoneural part were immunostained for both S1 and S2 peptides, while in the digestive system, the basiepithelial plexus and mucosal cell bodies were immunoreactive. These immunocytochemical data support the notion that the SALMFamides may play a neuroendocrine role in mediating feeding behaviour of the starfish. Further studies including identification of peptide binding sites and differential expression pattern of mRNAs encoding the peptides are required to elucidate their physiological functions.

• The Korean Journal of Zoology
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• v.30 no.2
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• pp.154-166
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• 1987

Recently, the researches on the enteroendocrine cells of vertebrates have made a remarkable advance by the immunocytochemical methods. This study was attempted to investigate the topographical distributions and the shapes of gastrin, glucagon, somatostatin and cholecystokinin-8 immuno-reactive cells in the gastrointestinal tract of the Korean hedgehog, Erinaceus koreanus. For light-microscopical examination of immunocytochemistry, the tissue specimens taken from the various portions(body and pyloric protion of stomach, duodenum, jejunum, ileum and rectum) were fixed in glutaraldehyde-picric acid-acetic acid (GPA) or 10% neutral buffered formalin solutions. For the demonstration of immunoreactive cells, the paraffin sections (6$\mu$m) were immunocytochemically identified by PAP procedure (Sternberger, 1979) with gastrin, glucagon, somatostatin and cholecystokinin-8 antisera. Gastrin-immunoreactive cells were mainly distributed in the pyloric portion of stomach and were a few in the duodenum and jejunum. The shapes of these cells were round or oval in the pyloric portion and pyramidal in the small intestine. Glucagon-immunoreactive cells were sparsely distributed in the only small intestine. The shapes of these cells were mainly pyramidal. Somatostatin-immunoreactive cells were a few in the pyloric portion and duodenum, and were sparsely distributed in the body of stomach and jejunum. The shapes of these cells were round or oval in the stomach and oval or pyramidal in the small intestine. Cholecystokinin-8-immunoreactive cells were sparsely distributed in the only small intestine. The shapes of these cells were mainly oval or pyramidal.

• Journal of Life Science
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• v.5 no.1
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• pp.1-7
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• 1995

Mammary orgamoids(ductal and endbud fragments) were cultured in a complete hormone medium(CHM) with 10%FBS, estradiol, progesterone, hydrocortisone, insulin, and prolactin, Several types of colonies were observed: stellate(14$$\pm$5.5%), duct(41$\pm$5.6%), web(35$\pm$3.6%), squamous(6$\pm$2.1%), and lobuloduct(4$\pm$1.2%), Squamous colony was typical squamous metaplasia(SM) with several layers of squamous epithlia and keratin pearls. At the immunocytochemical study, casein proteins were predominantly localized near the apical surfaces of the cells or in the lumina of ductal or lobuloductal colonies. To inhibit the formation of SM, we treated organoids with all-trans retinoic acid(RA) from 10$^{-6}$ to 10$^{-17}$ M in CHM. Formation of SN was completely inhibited at 10$^{-9}$M RA in CHM. The frequency of lobuloductal colony formation was increased with the augmentation of RA concentration.

• Applied Microscopy
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• v.25 no.1
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• pp.75-85
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• 1995

The goal of the present study is to investigate the precise variation of tubulin substances in the cytoplasm of oviductal ciliated cells and the morphological changes in cytoplasmic organelles of ciliated cells for ciliogenesis by estrous cycles. The animals used in this study were female rat (Sprague Dawley strain), weighing approximately 200 gm. The ampulla oviducts of these animals (at each of estrous cycle) were rapidly excised. At each stage of estrous cycle, the tissues were used for immunocytochemical study and other were used for electron microscopical study. All specimens were observed by the light and electron microscope. The results obtained are as follows: 1. In the ciliated cells at proestrus, Golgi complex showed $5{\sim}7$ stacked cisternae with numerous saccules and vacuoles. Large amount of fibrous granules were located near the Golgi complex. But at metestrus and diestrus, few fibrous granules were seen. 2. A moderate number of rough endoplasmic reticulum and polyribosomes were scattered in the cytoplasm of ciliated cells at proestrus, but were decrease in number at metestrus and diestrus. 3. At proestrus and estrus, there were a large amount of vesicles in the apical cytoplasm of ciliated cells. 4. Numerous mitochondria were located in the apical cytoplasm at proestrus and estrus, but only a few at metestrus and diestrus. 5. At proestrus and estrus, tubulin substances showed strong reactions in the cytoplasm but weak reactions at metestrus and diestrus. It is suggested consequently that the ciliated cells of the rat oviducts showed no morphological changes of cilia but the ultrastructural organelles of the cells were changed in its shape and location during the entire estrous cycle.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.6
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• pp.495-505
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• 2006

In this study we evaluate the effects of bFGF-BB and PDGF on in vitro proliferation, differentiation and mineralization of mesenchymal stem cells (MSCs) from rat. MSCs were prepared from the bone marrow of 6 or 7-week-old male rats with a technique previously described by Maniatopoulos et al. in 1988. Lineage differentiation to osteogenesis, chondrogenesis and adipogenesis were performed. At first, we characterized the cultured cell on passage 1, 3, 5, 7 with immunocytochemical staining using CD29, 44, 34, 45, ${\alpha}$-SMA and type I collagen. And to study the effects of bFGF and PDGF-BB on proliferation, differentiation and mineralization, we seeded the expanded cell at a density of 6 $6{\times}10^3\;cells/cm^2$ to 100-mm dish for evaluation of cell proliferation and MTT assay was carried out on day 2, 4, 7, 9. We also resuspended the cells with same density $(6{\times}10^3\;cells/cm^2)$ to 24 well plates for subculture. On the following day, the attached cells were exposed to 2.5ng/ml bFGF and/or 25ng/ml PDGF-BB daily during 5 days. The osteocalcin (OC) level was assessed and mineral contents were evaluated with alizarin red S staining on subculture day 2, 7, 14, 21. We identified the mesenchymal stem cell from the bone marrow derived cells of rat through their successful multi-differentiation and stable display of its phenotype. And bFGF and PDGF-BB showed the effect that inhibited osteoblastic differentiation and mineralization mildly in above concentration at in vitro culture. This study was supported by grant 04-2004-0120 from the Seoul National University Hospital Research Fund.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.7
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• pp.962-967
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• 2003

The purpose of the study were to characterize the proteins secreted by elongating caprine conceptus, to identify a group of low molecular weight proteins as retinol-binding protein (RBP), to identify RBP cell-specific localization in conceptus tissue, and to demonstrate that the conceptuses secreted continuously RBP during the time period maternal recognition of pregnancy. Caprine conceptuses were removed from the uterus between days 16 and 22 of pregnancy, the time period maternal recognition of pregnancy. Isolated conceptuses were cultured in a modified minimum essential medium in the presence of radiolabeled amino acids. Proteins synthesized and secreted into medium were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis and fluorography. At least five proteins showed consistently a grouping of spots with characteristic location on two-dimensional gels. A major low molecular weight protein consisted of two major isoforms (pI 5.3-6.0) of similar molecular mass (21 kDa) was identified as RBP by using antiserum against RBP. Presence of RBP in conceptus culture medium and uterine flushings between days 16 and 22 of pregnancy were determined by immunoprecipitation and Western blotting using anti-RBP serum. In immunocytochemical study, strong immunostaining for RBP was localized in trophectoderm and endoderm of conceptus. These results clearly demonstrated that the caprine conceptus was active in protein synthesis as early as day 16 of pregnancy. Secretion of RBP by caprine conceptuses (days 16-22) coincident with the rapid transformation of the conceptus from a spherical blastocyst to a filamentous structure. Production of RBP by the elongating conceptuses may be indicative of an important role for conceptus RBP in the transport, availability and metabolism of retinol during maternal recognition of pregnancy.

• Proceedings of the Korean Environmental Health Society Conference
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• 2005.06a
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• pp.341-344
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• 2005

The expression of the glutathione S-transferase (GST), whose induction accounts for antioxidant defense system, is regulated by activation of CCAAT/enhancer binding protein ${\beta}$ ($C/EBP{\beta}$), Sick house syndrome (SHS) presents healthy damage owing to the indoor environment of a building. Toluene has been implicated in one of the important causes of SHS. The present study investigated the effects of toluene treatment on the induction of GSTA2 gene and its mechanism. H411E cells treated with toluene, and GSTA2 expression was determined by immunoblot analysis. The translocation of $C/EBP{\beta}$ was assessed by immunocytochemical assays. $C/EBP{\beta}$ DNA binding activity was determined by electrophoretic mobility shift assays. The role of the C/EBP binding site in the induction of the GSTA2 gene was assessed by luciferase reporter-gene activity. Toluene induced GSTA2 protein expression. In toluene-treated cells, $C/EBP{\beta}$ translocated to the nucleus and bound to the consensus sequence of C/EBP (TTGCGCAA). Toluene treatment increased luciferase reporter-gene activity in cells transfected with the C/EBP-containing regulatory region of the GSTA2 gene. Oxidative stress is believed to play an important role in the induction of GSTA2 gene by toluene This study shows that toluene-induced GSTA2 gene expression is dependent upon nuclear translocation and binding of $C/EBP{\beta}$ to the C/EBP response element in the GSTA2 gene promoter.