• Journal of Physiology & Pathology in Korean Medicine
• /
• v.23 no.4
• /
• pp.805-817
• /
• 2009

This study was to investigate central localization of neurons projecting to the urinary bladder and urinary bladder-related acupoints(Wijung, $BL_{40}$) and neurons of immunoreactive to hormones and hormone receptors regulating urinary bladder function by using peudorabies virus(PRV). In this experiment, Bartha's strain of pseudorabies virus was used in rats to trace central localization of urinary bladder-related neurons and urinary bladder-related acupoints($BL_{40}$) which can regulate urinary system. PRV was injected into the urinary bladder and acupoints($BL_{40}$) related urinary system. After six days survival of rats, mainly common labeled neurons projecting to the urinary bladder and urinary bladder-related acupoints were identified in spinal cord, medulla, pons and diencephalon by PRV immunohistochemical staining method. First-order PRV labeled neurons projecting to urinary bladder and urinary bladder-related acupoints were found in the cervical, thoracic, lumbar and sacral spinal cord. Commonly labeled preganglionic neurons were labeled in the lumbosacral spinal cord and thoracic spinal cord. They were found in the lateral horn area(sacral parasympathetic nucleus and intermediolateral nucleus), lamina V-X, intermediomedial nucleus and dorsal column area. The area of sensory neurons projecting to urinary bladder and Wijung($BL_{40}$) was L5-S2 spinal ganglia and T12-L1 spinal ganglia, respectively. In the brainstem, the neurons were labeled most evidently and consistently in the nucleus of tractus solitarius, area postrema, dorsal motor nucleus of vagus nerve, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), C3 adrenalin cells, parapyramidal area(lateral paragigantocellular nucleus), locus coeruleus, subcoeruleus nucleus, A5 cell group, Barrington's nucleus and periaqueductal gray matter. In the diencephalon, PRV labeled neurons were marked mostly in the paraventricular nucleus and a few ones were in the lateral hypothalamic nucleus, posterior hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus, median eminence, perifornical nucleus, periventricular nucleus and suprachiasmatic nucleus. In cerebral cortex, PRV labeled neurons were marked mostly in the frontal cortex, 1,2 area, hind limb area, agranular insular cortex. Immunoreactive neurons to Corticotropin releasiing factor(CRF), Corticotropin releasiing factor-receptor(CRF-R), c-fos and serotonin were a part of labeled areas among the virus-labeled neurons of urinary bladder and Wijung($BL_{40}$). The commonly labeled areas were nucleus tractus solitarius, area postrema, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), locus coeruleus, A5 cell group, Barrington,s nucleus, arcuate nucleus, paraventricular nucleus, frontal cortex 1, 2 area, hind limb, and perirhinal(agranular insular) cortex. These results suggest that overlapped CNS locations are related with autonomic nuclei which regulate the functions of urinary bladder-relate organs and it was revealed by tracing PRV labeled neurons projecting urinary bladder and urinary bladder-related acupoints. These commonly labeled areas often overlap with the neurons connected with hormones and hormone receptors related to urination.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.21 no.2
• /
• pp.453-461
• /
• 2007

Drynariae Rhizoma has been used for promotes mending of the sinews and bone, tonifies the kidney for such symptoms as weak low back and knees, and stimulates the growth of hair as a tinctute for alopecia in oriental medicine. This experiment examined the effect of an acetone extracts of Drynariae Rhizomas(GSB-1), its EtoAc fraction(GB-2) and n-buOH fraction(GSB-3), on hair growth activity of the C57BL/6L mice after topical application to skin. First, We examined on hair growth activity of extracts of Drynariae Rhizomas compare to control and 1 % minoxidil groups. Second, We investigated on the number of hair follicle and mast cells after topical application of extracts of the Drynariae Rhizomas to skin for 16 day. Third, We investigated immunoreactive density of vascular endothelial growth factor(VEGF), protein kinase C-${\alpha}$(PKC-${\alpha}$) and stem(mast) cell factor(SCF) in skin of C57BL/6N mice by immunohistochemical methods. The results were as follows : Hair growth effect of acetone extracts of Drynariae Rhizomas, its EtoAc fraction and n-BuOH fraction was observed in 98 %, 96 % and 60 % in hair removed skin area in 16 day respectively, Immunoreactive density of VEGF in skin of GSB-1 group was weakly stained compare to control group in 10 day, But GSB-2 and GSB-3 groups were mildy stained in bulge and root sheath of skin. Immunolocalization of SCF antigens was observed weakly stained density in epidermis, bulge, stem cells and dermal papilla of control gruop. but in experimental group, immunoreactivity of SCF antigens was observed mildly stained density in bulge, epidermis and root sheath of GSB-1 gruop, heavily stained density in epidermis, bulge and root sheath of GSB-2 and GSB-3 groups to the hair removal skin of C57BL/6N mice on day 10. These experiment suggest that acetone extracts of Drynariae Rhizomas and its EtoAc fraction may be used for topical treatment of alopecia areata.

• Applied Microscopy
• /
• v.18 no.2
• /
• pp.59-76
• /
• 1988

In order to know the distribution, relative frequencies, types and morphology, endocrine cells in the intestinal tract of the hedgehog(Erinaceus koreanus) were studied by light microscopy, immunohistochemistry and electron microscopy. The results obtained are summarized as follows: 1. Two kinds of endocrine cells were demonstrated with two specific staining methods. Argyrophil cells(reactive cells for Grimelius method) were found most frequently in the intestinal region, and were infrequent in the rectum, whereas argentaffin cells (reactive cells for Masson-Hamperl method) were found most frequently in the rectum and in the other legions were infrequent. These reacting cells were mainly found in the intestinal glands, whereas a small number in the mucosa. 2. Twelve kinds of endocrine cells, gastrin(Gas)-, somatostatin (Som)-, serotonin(5-HT)-, glucagon(Glu)-, bovine pancreatic polypeptide(Bpp)-, cholecystokinin(Cck)-, secretin(Sec)-, motilin(Mot)-, glicentin(Gli)-, gastric inhibitor polypeptide(GIp)-, substance P(Sp)-, and neurotensin(Neu)-immunoreactive cells, were identified by immunohistochemical method. Gas-, Som-, 5-HT-, Glu-, Cck-, Sec-, Mot-, Gli-, Sp-, and Neu-reactive cells were observed in the duodenum, and among these Gas- and 5-HT-reactive cells were moderately found while the others were infrequent. In the jejunoileum Gas-, Som-, 5-HT-, Glu-, Cck-, Mot-, Gli-, GIp-, SP-, and Neu-reactive cells were found, and among these 5-HT- and GIp-reactive cells were moderately found while the others were infrequent. In the colon Sec-reactive cell was not detected. 5-HT-reactive cells were found most frequently and the others were infrequent in this region. 5-HT-, Bpp-, GIi- and Neu-reactive cells were found in the rectum. Among these 5-HT-reactive cells were found most frequently. 3. Electron microscopically, five types of endocrine cells, EC, ECL, D, G, A-like cell, were identified in the intestinal region. EC and ECL cells in the duodenum, EC, D and G cells in the jejuno-ileum, EC and A-like cells in the colon and EC cell in the rectum were observed respectively.

• The Korean Journal of Physiology and Pharmacology
• /
• v.7 no.4
• /
• pp.211-215
• /
• 2003

To examine the localization pattern of phospholipase D2 (PLD2) in the pancreatic islet (the islet of Langerhans) depending on species, we conducted a morphological experiment in the rat and guinea pig. Since individual islets display a typical topography with a central core of B cell mass and a peripheral boundary of A, D, and PP cells, double immunofluorescent staining with a panel of antibodies was performed to identify PLD2-immunoreactive cells in the islets PLD2 immunoreactivity was mainly present in A and PP cells of the rat pancreatic islets. And yet, in the guinea pig, PLD2 immunoreactivity was exclusively localized in A cells, and not in PP cells. These findings suggest a possibility that PLD2 is mainly located in A cells of rodent pancreatic islets, and that the existence of PLD2 in PP cells is not universal in all species. Based on these results, it is suggested that PLD2 may play a significant role in the function of A and/or PP cells via a PLD-mediated signaling pathway.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.79-79
• /
• 2003

Embryonic stem cells are capable of differentiating into a variety of cell lineages. However, the ultimate results of differentiation in vitro greatly depend on the duration of treatment and kinds of differentiating inducers added. In order to investigate the efficiencies of various differentiation inducers and the methods of treatment, we examined differentiation patterns of human embryonic stem cell (hESC, MB03) according to several different protocols. Exp. I) Upon differentiation using retinoic acid and ascorbic acid (RA/AA), embryoid bodies (EB, for 4days) derived from hESC was exposed to Rh (10$^{-6}$ M) and AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. Exp. III) In addition, to examine the effects of neurotrophic factors in the production of mature neurons, groups of cells were exposed to either BDNF (5 ng/ml) or TGF-$\alpha$(10 ng/ml) during the 28 days of final differentiation. Differentiation patterns of RA/AA or bFGF treated groups were very similar; approximately 82% and 83% of the cells, respectively, were positive for anti-NF200 antibody, while it was about 10% and 11%, respectively, for anti-NF160 antibody in 28 days in N2 medium. Alsor, cells expressing TH were as low as 5%, while the cells doubled when matured at the presence of either BDNF or TGF-$\alpha$. Cells immunoreactive to anti-GAD antibody were approximately 20%. These results suggest that a maturation step rather than differentiation induction step, which is formation of EB, effects more decisively to the ultimate differentiation pattern.

• Applied Microscopy
• /
• v.30 no.4
• /
• pp.377-387
• /
• 2000

There are a few tanycytes between the general ependymal cells lining the ependymal layer of the brain ventricle. These cells are considered as modified ependymal cells which possess a long basal process. Tanycytes are known to have an ability to communicate by absorbing substances from cerebrospinal fluid and transporting them to the blood vessels and/or to the neurons in the CNS. The third and fourth ventricular tanycytes were mainly studied as subjects but it's rare to find reports about the tanycytes of the area postrema. Glial fibrillary acidic protein is an intermediate filament protein that is expressed especially in astrocytes of the CNS. But GFAP is also found in filament of the tanycytes and its process. Therefore this study was carried out for the examination of the GFAP immunoreactive tanycytes lining the area postrema of the bat, and we also examined the ultrastructure of tanycytes using electron microscope. GFAP immunoreactive tanycytes were located in the caudal portion of the fourth ventricle, and especially mainly in the transitional zone between the floor of the caudal fourth ventricle and ependymal layer lining the area postrema. A few GFAP immunoreactive tanycytes were also found in the ependymal layer lining the area postrema, and some groups of tanycytes were found in the ependymal layer of the area postrema near the floor of the caudal fourth ventricle , The processes of tanycytes were stained deeply with anti-GFAP antibody. Especially the GFAP immunoreactive tanycytes lining the area postrema had very long processes that cross the whole width of the area postrema. In the electron microscope, the cell body of ependymal tanycyte was located on the ependymal layer and had a long basal process. Intermediate filaments were observed around the nucleus and well developed in the process of tanycrte. Longitudinal oriented long mitochondria and a few lipid droplets were also found in this process. After immunocytocheical staining, the gold particles were found only in the intermediate filaments. We could not determine the function of the tanycytes in the area postrema. Thus, further investigation is required to determine the functional relationship between the tanycytes and the area postrema in hibernating animal, the bat.

• Clinical and Experimental Reproductive Medicine
• /
• v.35 no.3
• /
• pp.181-191
• /
• 2008

Objective: Environmental chemicals alter reproduction, growth, and survival by changing the normal function of the endocrine system. Bisphenol A (BPA), one of the endocrine disruptors, is known to be an estrogen receptor agonist. Therefore, we hypothesized that BPA may affect male reproduction including spermatogenesis in the mouse testis. Methods: We used 7-week-old ICR mice. The first experiment group received BPA in sesame oil (vehicle, 1 mg/kg, 10 mg/kg, and 100 mg/kg) by i.p. injection and mice were sacrificed 24 hr later. The second experiment group received BPA (vehicle, 10 ${\mu}g/kg$, 1 mg/kg, and 100 mg/kg) daily for 14 days by subcutaneous injection. Expression of cell type-specific marker genes in the testis was evaluated by RT-PCR. Histological analysis, immunofluorescence staining, and TUNEL staining were also performed. Results: RT-PCR analyses showed that expression of luteinizing hormone receptor (LHR), a marker gene for the Leydig cell, was notably decreased in the testes of high dose-exposed mice. No obvious difference in the histology of testes was noted among treatment groups. Immunostaining of LHR in the first experiment group did not show noticeable difference in LHR protein expression in Leydig cells. Immunohistochemistry also revealed heightened expression of the immunoreactive Bax in the treatment group, and this was accompanied by positive TUNEL staining in the interstitial area within testis where Leydig cells reside. Conclusions: Our result suggests that BPA affects Leydig cell functions by altering gene expression and by increasing apoptosis in the mouse testis.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.32 no.3
• /
• pp.266-270
• /
• 1999

Two forms of gonadotropin releasing hormone (GnRH) are identified in the brain of adult mature spotted sea bass (Lateolabrax sp.) by immunohistochemical methods. Salmon GnRH immunoreactive (sGnRH-ir) cell bodies were distributed in the olfactory bulb, ventral telencephalon and preoptic region. Immunoreactive fibers were observed in the vicinity of the brain including the olfactory bulbs, the telencephalon, the optic nerve, the optic tectum, the cerebellum, the medulla oblongata and rostral spinal cord. In most cases, these fibers did not form well defined bundles. However, there was a clear continuum of immunoreactive fibers, extending from the olfactory bulbs to the pituitary. cGnRH-II-ir cell bodies were only found in olfactory bulbs. However, the distribution of cGnRH-II-ir fibers was basically similar to that of sGnRH-ir fibers except for the absence of their continuity between the olfactory bulbs and the pituitary. These data suggest that sGnRH and cGnRH-II are endogenous peptides and indicate the presence of multiple neuroendocrine functions in the brain of the spotted sea bass. It seems that sGnRH not only regulates GTH secretion but also functions as a neurotransmitter, whereas cGnRH-II functions only as a neurotransmitter.

• Applied Microscopy
• /
• v.32 no.2
• /
• pp.175-183
• /
• 2002

In this study, we carried out immunostaining and immunogold labeling with rabbit anti-dopamine (TH) and rabbit anti-calbindin-$D_{28K}$ to examine the characteristics and functions of the neurons that secrete neurotransmitters in optic lobes of Todarodes pacificus and Octopus minor inhabiting the Korean waters. The obtained results are as follow. In the immunostaining with anti-dopamine, only a few of the large amacrine cells in an the upper part of an outer granule cell layer and the cells forming the islands of medulla showed positive reaction in Todarodes pacificus, while $2{\sim}3$ cells in the upper and middle parts of an outer granule cell layer and more than 5 cells in the islands of medulla reacted positively in Octopus minor. For the case of anti-calbindin case, $2{\sim}3$ small amacrine cells in the upper portion of the outer granule cell layer and $1{\sim}2$ cells which are located in the lower part of an inner granule cell layer showed positive reaction in Todarodes pacificus, while, in Octopus minor, 4 cells in the outer granule cell layer reacted positively, no immunoreactive cell being found in the inner granule cell layer. As a result of performing the immunogold labeling, relative large number ($17{\sim}26$) of gold particles were labeled per $0.5{\mu}m^2$ of the cytoplasm of the cells which showed the immunoreactivity to the anti-dopamine and anti-calbindin in Todarodes pacificus, however, small number (10) of gold particles were labeled in Octopus minor, which reach only half of the number in the Todarodes pacificus.

• Asian Pacific Journal of Cancer Prevention
• /
• v.17 no.5
• /
• pp.2535-2537
• /
• 2016

Background: Most of the cervical cancer patients, including those with cervical adenocarcinomas, come at advanced stage in the developing world so its mortality is high. Radiotherapy is one of the treatment modality for advanced stage cervical adenocarcinomas, but its efficacy depends on several prognostic factors such as the stage, histopathology, presence of organ dysfunction and expression of cellular biology markers mainly involve in cell proliferation such as the epidermal growth factor receptor (EGFR). Some research indicates that activation of EGFR in malignancy (including cervical cancer) correlates with aggressive behavior, a poor prognosis and decreasing sensitivity of radiotherapy. However, the combination between targeted therapies and radiotherapy are innovative approaches which may provide a good result. This study aimed to assess any correlation between expression of EGFR and response to radiotherapy in cervical adenocarcinoma cases. Materials and Methods: A total of 32 women were registered in a retrospective study period January 2007 and May 2014. Paraffin blocks from these patients were processed by classical histological techniques and for immunohistochemical staining of EGFR, scoring being accomplished according to the immunoreactive scoring (IRS) of Remmele and Stegner. Results: Among the studied molecular factors, there was significant correlation expression of EGFR with poor response to radiotherapy (p=0.0001). Conclusions: The result of this study showed a significant correlation between expression of EGFR and sensitivity of radiation in cervical adenocarcinoma cases. Further research is necessary to obtain information about new therapeutic management.