• Title/Summary/Keyword: 3T3L1 cells

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Effect of the Combination of Co-Culture System and Supplemented Protein Sources on the In Vitro Development of Bovine IVF Embryos (각종 공동배양 배지와 첨가 단백질원의 조합이 소 체외수정란의 체외배양에 미치는 영향)

  • Cheong, H.T.;Lee, J.H.;Park, C.K.;Yang, B.K.;Kim, C.I.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.337-345
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    • 1999
  • The present study was conducted to investigate the effects of various co-culture systems and supplemented protein sources on the in vitro development of bovine IVF embryos. Bovine cumulus oocyte complexes (COCs) were matured and fertilized in vitro. Presumptive zygotes with cumulus cells were transferred to TCM-199 or CRlaa containing 10% FBS or 3mg/$m\ell$ BSA, and cultured for 36~40 hr. After primary culture, cleaved embryos were co-cultured with cumulus cells(CC), bovine oviduct epithelial cells(BOEC) or Buffalo rat liver cells (BRLC) in TCM-199 or CRlaa supplemented with FBS or BSA respectively, for further 6 days. Cleavage rate increased with BSA(P<0.01) in the both TCM-199(79%) or CRlaa(74%) When embryos were co-cultured with CC or BOEC in TCM-199, blastocyst development was enhanced with BSA(40% and 43%) compared to FBS (22% and 29%) , whereas in CRlaa no difference observed between BSA(40% and 39%) and FBS (40% and 42%). When embryos were co-cultured with BRLC monolayer, FBS enhanced the blastocyst development (P<0.05) compared to BSA in both TCM-199(41% vs 31%) and CRlaa (44% vs 37%). The result of the present study showed that the cleavage rate of bovine IVF embryos increased with BSA, The result also showed that BSA can enhance the development of IVF embryos in co-culture with CC or BOEC in TCM-199, suggesting the in vitro development is affected by the medium and supplemented protein sources in co-culture with somatic cells.

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Characterization of North American Ginseng Rust-Spot and the Effects of Ethephon

  • Campeau, Cindy;Proctor, John T.A.;Murr, Dennis P.;Schooley, Jan
    • Journal of Ginseng Research
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    • v.27 no.4
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    • pp.188-194
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    • 2003
  • Rust-spot on North American ginseng roots (Panax quinquefolius L.) is considered a physiological, not a pathological disorder. Ginseng rust-spot starts as an orange spot on the surface of the root and may spread forming a sunken, round to irregular lesion. 5 mm in diameter. Pieces of root, 7 mm in length and containing a rust-spotted lesion, were embedded in agar and sectioned using a vibratome. These sections and hand sections, cut with a two-sided razor blade, were examined using fluorescence microscopy. The 4-5 cell layers of the periderm were destroyed in the area of the lesion and orange substance:, were deposited in and around the lesion. Sections stained with vanillin-HCI and viewed using bright field microscopy confirmed that the orange substances were phenolic compounds. Scanning electron micros-copy showed that the periderm had pulled away from the root, or was completely destroyed, in the area of the lesion. The smooth surface of the lesion indicates the deposition of phenolic compounds in surrounding cells as a wound response. Roots sprayed or dipped in ethephon (1500 mgㆍL$^{-1}$ ) developed rust-spots, more so at 21$\pm$2$^{\circ}C$ than at 3$\pm$0.2$^{\circ}C$. Roots held at 21$\pm$2$^{\circ}C$ were yellowish and developed white cell proliferations. Comparable control roots also developed rust-spots likely due to the high undecomposed organic matter content of the incubation soilless mix.

Historical Review and Future of Cardiac Xenotransplantation

  • Jiwon Koh;Hyun Keun Chee;Kyung-Hee Kim;In-Seok Jeong;Jung-Sun Kim;Chang-Ha Lee;Jeong-Wook Seo
    • Korean Circulation Journal
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    • v.53 no.6
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    • pp.351-366
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    • 2023
  • Along with the development of immunosuppressive drugs, major advances on xenotransplantation were achieved by understanding the immunobiology of xenograft rejection. Most importantly, three predominant carbohydrate antigens on porcine endothelial cells were key elements provoking hyperacute rejection: α1,3-galactose, SDa blood group antigen, and N-glycolylneuraminic acid. Preformed antibodies binding to the porcine major xenoantigen causes complement activation and endothelial cell activation, leading to xenograft injury and intravascular thrombosis. Recent advances in genetic engineering enabled knock-outs of these major xenoantigens, thus producing xenografts with less hyperacute rejection rates. Another milestone in the history of xenotransplantation was the development of co-stimulation blockaded strategy. Unlike allotransplantation, xenotransplantation requires blockade of CD40-CD40L pathway to prevent T-cell dependent B-cell activation and antibody production. In 2010s, advanced genetic engineering of xenograft by inducing the expression of multiple human transgenes became available. So-called 'multi-gene' xenografts expressing human transgenes such as thrombomodulin and endothelial protein C receptor were introduced, which resulted in the reduction of thrombotic events and improvement of xenograft survival. Still, there are many limitations to clinical translation of cardiac xenotransplantation. Along with technical challenges, zoonotic infection and physiological discordances are major obstacles. Social barriers including healthcare costs also need to be addressed. Although there are several remaining obstacles to overcome, xenotransplantation would surely become the novel option for millions of patients with end-stage heart failure who have limited options to traditional therapeutics.

Herbal extract THI improves metabolic abnormality in mice fed a high-fat diet

  • Han, So-Ra;Oh, Ki-Sook;Yoon, Yoo-Sik;Park, Jeong-Su;Park, Yun-Sun;Han, Jeong-Hye;Jeong, Ae-Lee;Lee, Sun-Yi;Park, Mi-Young;Choi, Yeon-A;Lim, Jong-Seok;Yang, Young
    • Nutrition Research and Practice
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    • v.5 no.3
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    • pp.198-204
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    • 2011
  • Target herbal ingredient (THI) is an extract made from two herbs, Scutellariae Radix and Platycodi Radix. It has been developed as a treatment for metabolic diseases such as hyperlipidemia, atherosclerosis, and hypertension. One component of these two herbs has been reported to have anti-inflammatory, anti-hyperlipidemic, and anti-obesity activities. However, there have been no reports about the effects of the mixed extract of these two herbs on metabolic diseases. In this study, we investigated the metabolic effects of THI using a diet-induced obesity (DIO) mouse model. High-fat diet (HFD) mice were orally administered daily with 250 mg/kg of THI. After 10 weeks of treatment, the THI-administered HFD mice showed reduction of body weights and epididymal white adipose tissue weights as well as improved glucose tolerance. In addition, the level of total cholesterol in the serum was markedly reduced. To elucidate the molecular mechanism of the metabolic effects of THI in vitro, 3T3-L1 cells were treated with THI, after which the mRNA levels of adipogenic transcription factors, including C/$EBP{\alpha}$ and $PPAR{\gamma}$, were measured. The results show that the expression of these two transcription factors was down regulated by THI in a dose-dependent manner. We also examined the combinatorial effects of THI and swimming exercise on metabolic status. THI administration simultaneously accompanied by swimming exercise had a synergistic effect on serum cholesterol levels. These findings suggest that THI could be developed as a supplement for improving metabolic status.

Effect of lonizing Radiation on the Host Resistance Against Listeria Monocytogenes Infection and the Cytokine Production in Mice (방사선조사후 마우스에서의 Cytokine 생산능 및 Listeria monecytogenes에 대한 저항성의 변화)

  • Oh, Yoon-Kyeong;Chang, Mee-Young;Kang, In-Chol;Oh, Jong-Suk;Lee, Hyun-Chul
    • Radiation Oncology Journal
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    • v.15 no.3
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    • pp.175-186
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    • 1997
  • Purpose : To evaluate the qualitative immunologic changes by ionizing radiation. we studied the altered capacities of the macrophages and lymphocytes to produce cytokines in conjunction with resistance to Listeria monocytegenes (LM) infection in mice Materials and Methods : BALB/c mice and Listeria monocytogenes were used. The mice were infected intraperitoneally with $10^5LM$ at 1 day after irradiation (300cGy) and sacrificed at 1, 3, 5 days after infection, and then the numbers of viable LM per spleen in the irradiated and control group were counted. Tumor necrosis factor-alpha ($TNF-\alpha$), interferon-gamma ($IFN-\gamma$). interleukin-2 (IL-2), and nitric oxide (NO) were assessed after irradiation. Results : Under gamma-ray irradiation with a dose range of 100-850cGy, the number of total splenocytes decreased markedly in a dose-dependent manner, while peritoneal macrophages did so slightly Cultured peritoneal macrophages produced more $TNF-\alpha$ in the presence of lipopolysaccharide (LPS) during the 24 hours after in vitro irradiation, but their capacity of $TNF-\alpha$ Production showed a decreased tendency at 5 days after in vivo total body irradiation. With 100cGy and 300cGy irradiation, cultured peritoneal macrophages produced more NO in the presence of LPS during the 24 hours after in vitro irradiation than without irradiation. Activated splenocytes from irradiated mice (300cGy) exhibited a decreased capacity to Produce IL-2 and $IFN-\gamma$ with Concavalin-A stimulation at 3 days after irradiation. When BALB/c mice were irradiated to the total body with a dose of 300cGy, they showed enhanced resistance during early innate phase, but a significant inhibition of resistance to LM was found in the late innate and acquired T-cell dependent phases. Conclusion : These results su99es1 that increased early innate and decreased late innate and acquired immunity to LM infection by ionizing radiation (300cGy) may be related to the biphasic altered capacity of the macrophages to produce $TNF-\alpha$ and the decreased capacities of the lymphocytes to produce IL-2 and $IFN-\gamma$ in addition to a marked decrease in the total number of cells.

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Studies on the Relationship of the Central Neural Pathways to the Urinary Bladder and Wijung($BL_{40}$) (방광(膀胱)과 위중(委中)의 중추신경로와의 연계성에 관한 연구)

  • Lee, Chang-Hyun;Kim, Ho;Lee, Kwang-Gyu;Jeong, Han-Sol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.4
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    • pp.805-817
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    • 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.

A Study of Morphology and Pattern Analysis in Magnoliae Cortex (후박(厚朴)의 외.내부형태 및 이화학패턴연구)

  • Lee, Guem-San;Kim, Jung-Hoon;Choi, Go-Ya;Kang, Dae-Hoon;Hwang, Sung-Yeoun;Jeong, Seung-Il;Kim, Hong-Jun;Ju, Young-Sung
    • The Korea Journal of Herbology
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    • v.23 no.4
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    • pp.21-29
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    • 2008
  • Objectives: To determine the standards for discrimination of Magnoliae Cortex, the experiment of specific external-internal characters and the physicochemical pattern analysis were performed. Methods: External characteristics was observed using a stereoscope. Paraffin-mediated sectioned materials were stained by Ju's method. Physicochemical patterns of materials were analyzed using HPLC. Results: 1. Botanical characteristics: Magnolia officinalis had one seed and a white flower, while M. obovata had two seeds and a white flower. Machilus thunbergii had berry and spherical fruits and yellowish green panicles. 2. External characteristics: M. officinalis and M obovata were dark and thick. M. officinalis was gray brown and greasy while M. obovata was light-gray, less oily and smoothly sectioned. Machilus thunbergii was thin and relatively light or yellow-brown, coarsely sectioned and faintly specific scents. 3. Internal characteristics: The bast parts of M. officinalis and M. obovata were commonly wider than Machilus thunbergii The cork cortex of M. officinalis was $10{\sim}mg/L$ cell layers with many oil cells, while that of M. obovata was $4{\sim}7$ cell layers with less oil cells. Machilus thunbergii's xylem which consisted of ring-shaped cambium at 1st and 2nd part was occupied in large portion. 4. Physicochemical pattern: Both M. officinalis and M. obovata involved honokiol and magnolol. All kinds of M. officinalis involved Magnatriol B but one kind of M. ovobata and all of Machilus thunbergii didn't. Machilus thunbergii showed different pattern of chromatogram from that of 2 species above. Conclusions: These results could be used as standards for discrimination of Magnoliae Cortex and as the method of objectification in medicinal herbs giving the basic resource for bioactivity research.

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Pathogenicity of Salmonella gallinarum isolated from chickens in Korea (국내 분리주 Salmonella gallinarum의 닭에 대한 병원성)

  • Lee, Hee-soo;Kim, Soon-jae;Kim, Ki-seuk;Mo, In-pil;Kim, Tae-jong
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.569-576
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    • 1997
  • Fowl typhoid(FT) caused by Salmonella gallinarum is an infectious, egg-transmitted disease and characterized by swollen bronze liver, greenish-yellow diarrhea and high mortality in growing and adult chickens. Since 1992 the outbreak of FT has been increased. Several problems have been occurred such as absence of appropriate vaccines and lack of useful therapeutic methods. In these studies we investigated the pathogenicity of S gallinarum isolated in chickens. To compare the pathogenicity among the species of chickens, all chickens were challenged intramuscularly or orally with $1{\times}10^7$ CFU of S gallinarum. The brown-colored layers were more susceptible and white leghone chickens were more resistant than other species. In the brown layer chickens orally challenged, lethal doses ($LD_{50}$) of the isolates were inoculated at 1 day, 2 weeks, 4 weeks and 8 weeks old chickens with amount of $10^{4.2}$, $10^{4.7}$, $10^{7.0}$ and $10^{7.6}$ CFU, respectively. The chickens which were intramuscularly challenged with the less amount than $10^2$ CFU showed higher mortality than that of the chickens orally inoculated with same dose. Also, we investigated the recovery rates of bacteria from various organs of survival chickens which were challenged orally with $5{\times}l0^7$ CFU of S gallinarum. The bacteria was more frequently and isolated earlier from the liver and spleen than from any other ogans. In the pathogenicity test, the white-leghorn chickens which were known as resistant-strain against Salmonella were artificially immunosuppressed using bursectomy and/or dexamethasone treatment. Mortality of chickens with both bursectomized and treated with dexamethasone was higher(90%) than that of the control group(10%), the bursectomized chickens(10%) and the dexamethasone only treated group(20%). It was suggested that the protective mechanism in chickens against S gallinarum may be required both the functions of B-cells and T-cells.

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Induction of Apoptosis by Ethanol Extract of Lythrum anceps (Koehne) Makino in Human Leukemia U937 Cells (인체백혈병 U937 세포에서 부처꽃 에탄올추출물에 의한 apoptosis 유도)

  • Jeong, Jin-Woo;Kim, Chul Hwan;Lee, Young-Kyung;Hwang, Yong;Lee, Ki Won;Choi, Kyung-Min;Kim, Jung Il
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.279-286
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    • 2020
  • Purple loosestrife-Lythrum anceps (Koehne) Makino is a herbaceous perennial plant belonging to the Lythraceae family. It has been used for centuries in Korea and other Asian traditional medicine. It has been showed pharmacological effects, including anti-oxidant and anti-microbial effects. However, the mechanisms underlying its anti-cancer effect are not yet understood. In this study, we investigated the mechanism of apoptosis signaling pathways by ethanol extract of Lythrum anceps (Koehne) Makino (ELM) in human leukemia U937 cells. Treatment with ELM significantly inhibited cell growth in a dose-dependent manner by inducing apoptosis, as evidenced by the formation of apoptotic bodies (ApoBDs), DNA fragmentation and increased populations of sub-G1 ratio. Induction of apoptosis by ELM was connected with up-regulation of death receptor (DR) 4 and DR5, pro-apoptotic Bax protein expression and down-regulation of anti-apoptotic Bcl-2 protein, and inhibitor of apoptosis protein (IAP) family proteins, depending on dosage. This induction was associated with Bid truncation, mitochondrial dysfunction, proteolytic activation of caspases (-3, -8 and -9) and cleavage of poly(ADP-ribose) polymerase protein. Therefore, our data indicate that ELM suppresses U937 cell growth by activating the intrinsic and extrinsic apoptosis pathways, and thus may have applications as a potential source for an anti-leukemic chemotherapeutic agent.

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.