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Effect of Imipramine on the Contractility of Single Cells Isolated from Canine Detrusor (Imipramine이 배뇨근 세포의 수축성에 미치는 직접작용)

  • Huh, Chan-Wook;Lee, Kwang-Youn;Kim, Won-Joon
    • Journal of Yeungnam Medical Science
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    • v.11 no.2
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    • pp.293-302
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    • 1994
  • The objective of this study was to establish a good methodology to isolate single smooth muscle cells that are alive and respond properly to pharmacological agents. Canine urinary bladders were employed as the source of single cells, and acetylcholine, atropine and imipramine were used as indicators of pharmacological responsiveness. Imipramine, an antidepressant drug exhibited the anticholinergic and calcium antagonizing properties on rat detrusor muscle. To establish a control value for a further experiment to elucidate the mechanism of action of imipramine on detrusor muscle, we measured the concentration-response of single cells to acetylcholine in the presesnce of imipramine by length of the cells and compared the result with the response in the presence of atropine. Tiny chops of smooth muscle taken from anesthetized canine urinary bladder were incubated in collagenase solution at $36^{\circ}C$ for 17-20 minutes. The collagenase solution included collagenase 1.2 mg/ml, soybean tryspin inhibitor 0.08 mg/ml, bovine serum albumin 2% in 10 ml Krebs-Henseleit buffer solution aerated with a consistent breeze of 95/5% $O_2/CO_2$, to maintain the pH at 7.4. After washing with plain K-H solution on 450 mesh, cells were dissociated from the digested tissue for 12-15 minutes. Cell suspension was transfered in 5 ml test tubes and acetylcholine was added for the final concentration to be $10^{-14}M{\sim}10^{-9}M$. To find the optimal time to fix the cells to determine the contractile responses, 1% acrolein was added 5, 10, 20, 30, 60 and 120 seconds after the administration of ACh. The length of cells fixed by acrolein were measured by microscaler via CCTV camera on phaes-contrast microscope. The average length of 50 cells from a slide glass was taken as the value of a sample at the very concentration point. Single cells were isolated from canine detrusor. The length of untreated cells varied from 82 ${\mu}m$ to 94 ${\mu}m$. The maximal response to actylcholine $10^{-9}M$ was accomplished within 5 seconds of exposure, and the shortening was $19{\pm}3$%. Atropine reduced the contraction of the cells concentration-dependently. Imipramine which exerts a cholinergic blocking action on some smooth muscles also reduced the contraction concentration-dependently and by a similar pattern as atropine. These findings document that imipramine may exerts a cholinergic blocking activity in the single smooth muscle cells isolated from canine urinary bladder.

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Mucin2 is Required for Probiotic Agents-Mediated Blocking Effects on Meningitic E. coli-Induced PathogenicitiesS

  • Yu, Jing-Yi;He, Xiao-Long;Puthiyakunnon, Santhosh;Peng, Liang;Li, Yan;Wu, Li-Sha;Peng, Wen-Ling;Zhang, Ya;Gao, Jie;Zhang, Yao-Yuan;Boddu, Swapna;Long, Min;Cao, Hong;Huang, Sheng-He
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1751-1760
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    • 2015
  • Mucin2 (MUC2), an important regulatory factor in the immune system, plays an important role in the host defense system against bacterial translocation. Probiotics known to regulate MUC2 gene expression have been widely studied, but the interactions among probiotic, pathogens, and mucin gene are still not fully understood. The aim of this study was to investigate the role of MUC2 in blocking effects of probiotics on meningitic E. coli-induced pathogenicities. In this study, live combined probiotic tablets containing living Bifidobacterium, Lactobacillus bulgaricus, and Streptococcus thermophilus were used. MUC2 expression was knocked down in Caco-2 cells by RNA interference. 5-Aza-2'-deoxycytidine (5-Aza-CdR), which enhances mucin-promoted probiotic effects through inducing production of Sadenosyl-L-methionine (SAMe), was used to up-regulate MUC2 expression in Caco-2 cells. The adhesion to and invasion of meningitic E. coli were detected by competition assays. Our studies showed that probiotic agents could block E. coli-caused intestinal colonization, bacteremia, and meningitis in a neonatal sepsis and meningitis rat model. MUC2 gene expression in the neonatal rats given probiotic agents was obviously higher than that of the infected and uninfected control groups without probiotic treatment. The prohibitive effects of probiotic agents on MUC2-knockdown Caco-2 cells infected with E44 were significantly reduced compared with nontransfected Caco-2 cells. Moreover, the results also showed that 5-Aza-CdR, a drug enhancing the production of SAMe that is a protective agent of probiotics, was able to significantly suppress adhesion and invasion of E44 to Caco-2 cells by upregulation of MUC2 expression. Taken together, our data suggest that probiotic agents can efficiently block meningitic E. coli-induced pathogenicities in a manner dependent on MUC2.

The Improvement of Chaga Mushroom (Inonotus Obliquus) Extract Supplementation on the Blood Glucose and Cellular DNA Damage in Streptozotocin-Induced Diabetic Rats (Streptozotocin으로 유발한 당뇨쥐에 있어서 차가버섯(Inonotus Obliquus)의 혈당 및 DNA 손상 개선효과)

  • Park, Yoo-Kyoung;Kim, Jung-Shin;Jeon, Eun-Jae;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.42 no.1
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    • pp.5-13
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    • 2009
  • Mushrooms have become a largely untapped source of powerful new pharmaceutical products that poses anti-inflammatory, and antimutagenic, and antioxidant activities. The antioxidant effects of the mushroom may be partly explained by protecting cellular components against free radical. The aim of this study was to investigate the protective effect of chaga mushroom against diabetes, via the mitigation of oxidative stress and reduction of blood glucose, in streptozotocin-induced diabetic rats. Rats were rendered diabetic by intravenous administration of STZ through tail at a dose of 50 mg/kg. Animals were allocated into four groups with 8 rats each. The control and diabetic control group were fed with standard rat feed. The other diabeic groups, the low chaga extract group and the high chaga extract group were fed ad libitum using 0.5 g/kg and 5 g/kg of chaga mushroom extract, respectively, for 4 weeks. The blood glucose levels in the two chaga extract groups showed a tendency to decrease but did not reach statistical significance after the supplementation. Leukocyte DNA damage, expressed as tail length, was found to be significantly lower in the high chaga extract group than in the diabetic control group (p > 0.05). Plasma level of total radical-trapping antioxidant potential (TRAP) was tend to be higher in the high chaga extract group compared with the diabetic control group. Erythrocyte antioxidant enzyme activities of two groups did not differ. Although we did not obtain beneficial effect on lowering blood glucose levels in the STZ-induced diabetic rats, this results suggest that the chaga mushroom extracts may initially act on protecting endogenous DNA damage in the short-term experiment.

Policy Study on Korean Retail Micro Business (국제 비교를 통한 소매업 소상공인 현황과 정책적 시사점)

  • Suh, Yong Gu;Kim, Suk Kyung
    • Journal of Distribution Research
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    • v.17 no.5
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    • pp.39-57
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    • 2012
  • The unabated influx of micro businesses has turned the Korean retailing market to a rat race, which causes severe financial distress for micro business owners due to heavy competition. The woes of these micro business owner's are exacerbated by the presence of large scale distributors such as Super Supermarket(SSM) and large discount stores. In summary, the Korean retail market is overburdened an uneconomically viable. Retailing has low barriers to entry which attracts unskilled labor or those with little capital. These start-ups have low opportunity costs since they would make low wages elsewhere in the economy. Thus, these owners are content with relatively low returns on their investment. These 'subsistence ventures' are maintained for economical viability rather than economic growth. These 'subsistence ventures' intensifies competition among small-scale businesses. The presence of large retail corporations also aggravates the situation. The recent stagnation of the economy has worsened the retail market in Korea. The overwhelming competition solidifies the coarse structural system and the prolonged economic sluggishness has increased the risk of insolvency for micro business owners. As the economy continues to stagnate, the imminent risk in retailing market will rise up to surface threatening economic stability. More systematic inflows and outflows of retailers are required in order to redress this structural problem. It has been empirically shown that the self-employment rate is high in Korea compared to other OECD countries. To draw the comparison of self-employment rate by industry, Korea shows high rates among transportation, whole sale, retail, education, lodging, and restaurants. In the case of the transportation and education service sectors, this high rate can be explained by the idiosyncratic nature of Korean culture. In the transportation sector, political policies favor private cap service and private freight carriers. In the education service sector, Koreans put particular emphasis on education that leads to many private institutions that outnumber other OECD countries. For these singular reasons, Korea maintains high micro business, self-employed rates particularly in retailing. A comparable nation is Japan, with its similar social, economic, cultural environment among OECD countries. Unlike Korea, Japan has much lower rates of micro business which continues to decrease. Also Korean retailers are much more destitute than Japanese. The fundamental problem of Korean retailing is the involuntary exit of these 'subsistence ventures,' micro businesses with low margins, in which a small drop in demand can lead to financial difficulties for the owner. This problem will be exacerbated when Korean babyboomers retire and join the micro business ventures. The first priority in order to cope with the severity of oversupply in retailing is to provide better opportunities for the potential self-employers. There should be viable alternatives to subsistent ventures. Strengthening the retirement program, scrutiny of exit process, reconfiguration of policy funds are the recommendations.

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Effect of Low Molecular Alginates on Cholesterol Levels and Fatty Acid Compositions of Serum and Liver Lipids in Cholesterol-Fed Rats (저분자화알긴산이 콜레스테롤식이 흰쥐의 혈청과 간장지질의 콜레스테롤 수준 및 지방산 조성에 미치는 영향)

  • LEE Dong-Soo;NAM Taek-Jeong;PYEUN Jae-Hyeung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.3
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    • pp.399-408
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    • 1998
  • The effect of low molecular alginates feeding on the cholesterol levels and fatty acid compositions of rat serum and liver lipid were investigated. After one week basal diet feeding, four week old Sprague-Dawley male rats were fed with water soluble and acid $\cdot$alkali soluble alginate extracted from sea mustard (Undaria pinnatifida) and giant kelp (Macrocystis pyrifera), and their low molecular alginates prepared by the HCl partial hydrolysis. The feeding efficiency of the alginate fed group was ranged in 0.37$\~$0.44, which was 0.03$\~$0.05 lower than that of the basal diet group. Also, there was much less increase of liver weight in the alginate fed group. The water soluble alginate showed more significant effect in reducing the total cholesterol, free cholesterol, LDL-cholesterol, triglyceride and phospholipid of serum and liver lipid than the acid$\cdot$alkali soluble alginate. The effect was much better with low molecular alginate (reducing effect by the low-molecularization : Water soluble alginate - serum lipid; total cholesterol $59\%$, free cholesterol $65\%$, LDL-cholesterol $96\%$, triglyceride $50\%$, and phospholipid $36\%$. liver lipid: total cholesterol $4\%$, free cholesterol $62\%$, LDL-cholesterol $44\%$, triglyceride $33\%$, and phospholipid $44\%$. acid$\cdot$alkali soluble alginate - serum lipid; total cholesterol $52\%$: free cholesterol $97\%$, LDL-cholesterol $78\%$ triglyceride $32\%$, and phospholipid $64\%$. liver lipid; total cholesterol $11\%$, free cholesterol $12\%$, LDL-cholesterol $10\%$, triglyceride $27\%$, and phospholipid $21\%$). The effect of low molecular alginate feeding on the fatty acid composition of serum and liver lipid reflects the remarkable increase of polyenoic acid, over $44\%$ in serum lipid and about $70\%$ in liver lipid, comparing to the cholesterol fed group. The overall results indicated that feeding of low molecular alginates improves physiological function of rats by changing the serum and liver lipid composition.

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Effects or $H_2O$ Fraction or Dioscorea japonica Thunb with selenium on plasma Glucose and Lipid Metabolism in Streptozotocin Induced Diabetic Rats (당뇨 흰쥐에서의 참마 $H_2O$ 분획물과 Selenium 보충시 혈당 및 지질대사에 미치는 영향)

  • 김명화
    • Journal of Nutrition and Health
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    • v.31 no.9
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    • pp.1377-1384
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    • 1998
  • The purpose of this study was to investigate the effect of H$_2$O fraction of Dioscorea japonica Thunb(DJT) with selenium(Se) treatment on blood glucose and lipid metabolism in streptozotocin(STZ)-induced diabetic rats. Male Sprague-Dawley rats(180-220g) weighing were divided into five groups, that is one normal group and four diabetic experimental groups : the STZ-control group, the DJT group, the DJT-Se group and the Se group. Diabetes mellitus was induced in the male rats by injection of STZ into the tail vein at a dose of 45mg/kg B.W. The H,0 fraction of DJT(500mg/kg) given orally were administered for 14 days. The Se treated group were fed a AIN-76 recommendation diet mixed with Na$_2$SeO$_3$(2mg/kg diet), which was prepared fresh daily. The body weight and food intake was monitored daily and plasma levels of glucose, insulin, hematocrit and protein were determined. The plasma concentrations of cholesterol, HDL-cholesterol, triglyceride and fire fatty acid were measured. The activities of aminotrans ferase were analysed. The body weight gain was shown to be significantly higher in the normal group than all diabetic groups. The blood glucose levels of the DIT-Se group was significantly lower compared to those of the experimental groups. The administration of H$_2$O fraction of DJT and selenium showed an increase in plasma protein concentrations. The plasma cholesterol levels of all STZ-groups were not significantly different and HDL-cholesterol levels were increased in the diabetic experimental groups fed on H$_2$O fraction of DJT or Se supplementation. Plasma triglyceride levels were lower in the DJT-Se and Se group than in the STZ-control group. free fatty acid levels were not significantly differ among STZ-control groups. STZ treatment increased aminotransferase activity and that of DJT group was highest. In conclusion, the data from the present experiments indicate that the treatment of the H$_2$O fraction of DJT with selenium showed a synergistic effect and the two can have an influence on hyperg1ycemia and lipid metabolites when administered together. (Korean J Nutrition 31(9) : 1377-1384, 1998)

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A Interpretation on Placeness and Historic Cultural Landscape of Dokjae-dong on Nogosan, Yangju (양주 노고산(老姑山) 독재동(篤才洞)의 장소성과 역사문화경관 해석)

  • Rho, Jae-Hyun;Kim, Hwa-Ok;Lee, Jung-Han;Park, Joo-Sung
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.33 no.1
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    • pp.97-109
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    • 2015
  • This study was focused on the placeness of Nogosan Dokjae-dong and rock inscription located in Jangheung-myeon, Yangju-si, Gyeonggi-do. The purpose was to reveal the implication of Nogosan and Dokjae-dong Valley in terms of Historic cultural landscape by considering the characteristics of the place and its landscape, and inferring the significance of the letters engraved on the rock. The result is as follows.10) Samgaksan is one of the five famous mountains in Korea. It is the sacred mountain that stands behind the capital city of the Joseon Dynasty that lasted for 600 years. On the other hand, the placeness of Nogosan is significant since it was the place where people used to bow low to the three great summits of Samgaksan. Furthermore, Nogosan was the perfect place to view Samgaksan that represented the Joseon spirit. Many letters were engraved on the rocks around Dokjae-dong Valley by Misu Heo Mok(許穆) as part of his memories to describe his experience in the mountain and became a scenery. Chusa(秋史) Kim Jung-Hee(金正喜) also engraved letters of 'Mongjae' on the rock that contains a double meaning. Literally, it means 'a dreaming house' However, it also implies that even just a daydream is significant in a way. These letters are presumed to be the signs that are related to Sung-Jae(性齋) Heo Jeon(許傳)'s experience in Dokjae-dong since the letters were part of his famous book title. Below these two letters, Chusa also engraved letters that describe the sexagenary cycle including the year of the rat, horse, dog, and tiger. They are regarded as the letters that describe the fate of Chusa's friend, a scholar born in May 1804. Nogosan and Dongjae-dong Valley are very significant in terms of historic cultural landscape as a viewpoint to see Samgaksan; and a place that cherishes the traces of Misu and shows common values and cohesiveness of well-known scholars of the Joseon Dynasty including Song Wol-Jae(松月齋) and Sung-Jae as a of Gihonamin(畿湖南人). The engraved letters of Chusa and Misu also implicitly demonstrates the major trends of Korea's calligraphy history.

Regulation of $LH{\beta}$ subunit mRNA by Ovarian Steroid in Ovariectomized Rats (난소제거된 흰쥐에서 난소호르몬에 의한 $LH{\beta}$ subunit의 유전자 발현조절)

  • Kim, Chang-Mee;Park, Deok-Bae;Ryu, Kyung-Za
    • The Korean Journal of Pharmacology
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    • v.29 no.2
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    • pp.225-235
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    • 1993
  • Pituitary LH release has been known to be regulated by the hypothalamic gonadotropin releasing hormone (GnRH) and the gonadal steroid hormones. In addition, neurotransmitters and neuropeptides are actively involved in the control of LH secretion. The alteration in LH release might reflect changes in biosynthesis and/or posttranslational processing of LH. However, little is known about the mechanism by which biosynthesis of LH subunits is regulated, especially at the level of transcription. In order to investigate if ovarian steroid hormones regulate the LH subunit gene expression, ${\alpha}\;and\;LH{\beta}$ steady state mRNA levels were determined in anterior pituitaries of ovariectomized rats. Serum LH concentrations and pituitary LH concentrations were increased markedly with time after ovariectomy. ${\alpha}\;and\;LH{\beta}$ subunit mRNA levels after ovariectomy were increased in a parallel manner with serum LH concentrations and pituitary LH contents, the rise in $LH{\beta}$ subunit mRNA levels being more prominent than the rise in ${\alpha}\;subunit$ mRNA. ${\alpha}\;and\;LH{\beta}$ subunit mRNA levels in ovariectomized rats were negatively regulated by the continuous treatment of ovarian steriod hormones for $1{\sim}4\;days$ and $LH{\beta}\;subunit$ mRNA seemed to be more sensitive to negative feedback of estradiol than progesterone. Treatment of estrogen antagonist, LY117018 or progesterone antagonist, RU486 significantly restroed LH subunit mRNA levels as well as LH release which were suppressed by estradiol or progesterone treatment. These results suggest that ovarian steroids negatively regulate the LH synthesis at the pretranslational level by modulating the steady state levels of ${\alpha}\;and\;LH{\beta}\;subunit$ mRNA and $LH{\beta}\;subunit$ mRNA seemed to be more sensitive to negative feedback action of estradiol than progesterone.

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Differential Function of EDRF in Systemic Arterial and Pulmonary Arterial System of Renal Hypertensive Rats (신성 고혈압쥐의 전신성 동맥계와 폐동맥계에 대한 EDRF 기능의 차이)

  • Lee, Byung-Ho;Shin, Hwa-Sup;Huh, In-Hoe
    • The Korean Journal of Pharmacology
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    • v.29 no.2
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    • pp.213-223
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    • 1993
  • To investigate the endothelium dependent vascular reactivity of the systemic arterial and the pulmonary arterial system in acute renal hypertensive rats of 2-kidney, 1-ligation type (RHRs), acetylcholine (ACh)-induced vasodilation and depressor effects were evaluated in isolated arteries and in vivo, respectively, in the presence and absence of functional endothelium. ACh $(10^{-5}\;M)$ relaxed the intact thoracic aortas from RHRs and normotensive rats (NRs), but the effect was significantly smaller for those from RHRs (34 and 86%, respectively, p<0.01). ACh-induced vasodilation was completely abolished after removal of endothelial cell or pretreatment with EDRF inhibitors, L-NAME and MB, indicative of its dependence on intact endothelial or EDRF function. ACh also induced vasorelaxation of the intact pulmonary arteries from RHRs and NRs; however, unlike the effects on the thorcic aorta, no significant difference in amplitude was noted between two groups. ACh $(0.1{\sim}10\;{\mu}g/kg,\;i.v.)$ reduced mean systemic arterial pressure in anesthetized RHRs and in NRs to the similar magnitude (% change: 39 and 46% at $10\;{\mu}g/kg$, respectively) and these hypotensive effects were significantly decreased after pretreatment with L-NAME (30 mg/kg, i.v.). Deprssor effects of ACh on mean pulmonary arterial pressure were similar in RHRs and NRs with and without pretreatment of L-NAME. However, in both NRs and RHRs, the depressor effects of ACh on mean pulmonary arterial pressure were significantly reduced compared with those for mean systemic arterial pressure, and the increment of mean pulmonary arterial pressure noted after L-NAME $(0.1{\mu}100\;mg/kg,\;i.v.)$ was significantly smaller than that for mean systemic arterial pressure. These results indicate that in RHRs the endothelial cell function was impaired, at least in part, in systemic arterial system, but not in pulmonary arterial system, and both ACh-evoked and basal release of EDRF was less in the pulmonary arterial system than in systemic arterial system of both NRs and RHRs.

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Neural pathway innervating ductus Deferens of rats by pseudorabies virus and WGA-HRP (흰쥐에서 WGA-HRP와 pseudorabies virus를 이용한 정관의 신경로에 대한 연구)

  • Lee, Chang-Hyun;Chung, Ok-Bong;Ko, Byung-Moon;Lee, Bong-Hee;Kim, Soo-Myung;Kim, In-Shik;Yang, Hong-Hyun
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.11-24
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    • 2003
  • This experimental studies was to investigate the location of PNS and CNS labeled neurons following injection of 2% WGA-HRP and pseudorabies virus (PRY), Bartha strain, into the ductus deferens of rats. After survival times 4-5 days following injection of 2% WGA-HRP and PRV, the rats were perfused, and their brain, spinal cord, sympathetic ganglia and spinal ganglia were frozen sectioned ($30{\mu}m$). These sections were stained by HRP histochemical and PRY inummohistochemical staining methods, and observed with light microscope. The results were as follows ; 1. The location of sympathetic ganglia projecting to the ductus deferens were observed in pelvic ganglion, inferior mesenteric ganglion and L1-6 lwnbar sympathetic ganglia. 2. The location of spinal ganglia projecting to the ductus deferens were observed in T13-L6 spinal ganglia. 3. The PRY labeled neurons projecting to the ductus deferens were observed in lateral spinal nucleus, lamina I, II and X of cervical segments. In thoracic segments, PRY labeled neurons were observed in dorsomedial part of lamina I, II and III, and dorsolateral part of lamina IV and V. Densely labeled neurons were observed in intermediolateral nucleus. In first lumbar segment, labeled neurons were observed in intermediolateral nucleus and dorsal commisural nucleus. In sixth lumbar segment and sacral segments, dense labeled neurons were observed in sacral parasympathetic nuc., lamina IX and X. 4. In the medulla oblongata, PRV labeled neurons projecting to the ductus deferens were observed in the trigeminal spinal nuc., A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nuc., rostroventrolateral reticular nuc., area postrema, nuc. tractus solitarius, raphe obscurus nuc., raphe pallidus nuc., raphe magnus nuc., parapyramidal nuc., lateral reticular nuc., gigantocellular reticular nuc.. 5. In the pons, PRV labeled neurons projecting to the ductus deferens were ohserved in parabrachial nuc., Kolliker-Fuse nuc., locus cooruleus, subcooruleus nuc. and AS noradrenalin cells. 6. In midbrain, PRV labeled neurons projecting to the ductus deferens were observed in periaqueductal gray substance, substantia nigra and dorsal raphe nuc.. 7. In the diencephalon, PRV labeled neurons projecting to the ductus deferens were observed in paraventricular hypahalamic nuc., lateral hypothalamic nuc., retrochiasmatic nuc. and ventromedial hypothalamic nuc.. 8. In cerebrum, PRV labeled neurons projecting to the ductus deferens were observed in area 1 of parietal cortex. These results suggest that WGA-HRP labeled neurons of the spinal cord projecting to the rat ductus deferens might be the first-order neurons related to the viscero-somatic sensory and sympathetic postganglionic neurons, and PRV labeled neurons of the brain and spinal cord may be the second and third-order neurons response to the movement of smooth muscles in ductus deferens. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitaing the internal environment. These observations provide evidence for previously unknown projections from ductus deferens to spinal cord and brain which may be play an important neuroanatornical basic evidence in the regulation of ductus deferens function.