• 제목/요약/키워드: Calcium imaging

검색결과 97건 처리시간 0.029초

Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

  • Han, Rafael Taeho;Kim, Han-Byul;Kim, Young-Beom;Choi, Kyungmin;Park, Gi Yeon;Lee, Pa Reum;Lee, JaeHee;Kim, Hye young;Park, Chul-Kyu;Kang, Youngnam;Oh, Seog Bae;Na, Heung Sik
    • The Korean Journal of Physiology and Pharmacology
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    • 제22권2호
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    • pp.173-182
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    • 2018
  • Recent studies have provided several lines of evidence that peripheral administration of oxytocin induces analgesia in human and rodents. However, the exact underlying mechanism of analgesia still remains elusive. In the present study, we aimed to identify which receptor could mediate the analgesic effect of intraperitoneal injection of oxytocin and its cellular mechanisms in thermal pain behavior. We found that oxytocin-induced analgesia could be reversed by $d(CH_2)_5[Tyr(Me)^2,Dab^5]$ AVP, a vasopressin-1a (V1a) receptor antagonist, but not by $desGly-NH_2-d(CH_2)_5[D-Tyr^2,Thr^4]OVT$, an oxytocin receptor antagonist. Single cell RT-PCR analysis revealed that V1a receptor, compared to oxytocin, vasopressin-1b and vasopressin-2 receptors, was more profoundly expressed in dorsal root ganglion (DRG) neurons and the expression of V1a receptor was predominant in transient receptor potential vanilloid 1 (TRPV1)-expressing DRG neurons. Fura-2 based calcium imaging experiments showed that capsaicin-induced calcium transient was significantly inhibited by oxytocin and that such inhibition was reversed by V1a receptor antagonist. Additionally, whole cell patch clamp recording demonstrated that oxytocin significantly increased potassium conductance via V1a receptor in DRG neurons. Taken together, our findings suggest that analgesic effects produced by peripheral administration of oxytocin were attributable to the activation of V1a receptor, resulting in reduction of TRPV1 activity and enhancement of potassium conductance in DRG neurons.

방사선조사가 MC3T3-E1 골모세포주의 TGF-${\beta}_1$ mRNA 발현과 석회화결절 형성에 미치는 영향 (Effects of irradiation on TGF-${\beta}_1$ mRNA expression and calcific nodule formation in MC3T3-E1 osteoblastic cell line)

  • 송주섭;김경아;고광준
    • Imaging Science in Dentistry
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    • 제38권3호
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    • pp.125-132
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    • 2008
  • Purpose : To investigate the effects of irradiation on transforming growth factor ${\beta}_1$ (TGF-${\beta}_1$) mRNA expression and calcific nodule formation in MC3T3-E1 osteoblastic cell line. Materials and Methods : Cells were cultured in alpha-minimum essential medium ($\alpha$-MEM) supplemented with 10% fetal bovine serum and antibiotics. When the cells reached the level of 70-80% confluence, culture media were changed with $\alpha$-MEM supplemented with 10% FBS, 5 mM $\beta$-glycerol phosphate, and $50\;{\mu}g/mL$ ascorbic acid. Thereafter the cells were irradiated with a single dose of 2, 4, 6, 8 Gy at a dose rate of 1.5 Gy/min. The expression pattern of TGF-${\beta}_1$ mRNA, calcium content and calcific nodule formation were examined on day 3, 7, 14, 21, 28, respectively, after the irradiation. Results : The amount of TGF-${\beta}_1$ mRNA expression decreased significantly on day 7 after irradiation of 4, 6, 8 Gy. It also decreased on day 14 after irradiation of 6, 8 Gy. and decreased on day 21 after irradiation of 8 Gy. The amount of calcium deposition decreased significantly on day 7 after irradiation of 4, 8 Gy (P < 0.01) and showed a decreased tendency on day 14, 21 after irradiation of 4, 6, 8 Gy. The number of calcific nodules was decreased on day 7 after irradiation of 4, 8 Gy. Conclusion: Irradiation with a single dose of 4, 6, 8 Gy influences negatively the bone formation at the molecular level by affecting the TGF-${\beta}_1$ mRNA expression that was associated with proliferation and the production of extracellular matrix in MC3T3-E1 osteoblastic cell line.

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Purinergic Receptors Play Roles in Secretion of Rat von Ebner Salivary Gland

  • Kim, Sang-Hee;Cho, Young-Kyung;Chung, Ki-Myung;Kim, Kyung-Nyun
    • International Journal of Oral Biology
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    • 제31권4호
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    • pp.141-148
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    • 2006
  • The effects of adenosine triphosphate(ATP) on salivary glands have been recognized since 1982. The presence of purinergic recepetors(P2Rs) that mediate the effects of ATP in various tissues, including parotid and submandibular salivary gland, has been supported by the cloning of receptor cDNAs and the expression of the receptor proteins. P2Rs have many subtypes, and the activation of these receptor subtypes increase intracellular $Ca^{2+}$, a key ion in the regulation of the secretion in the salivary gland. The apical pores of taste buds in circumvallate and foliate papillae are surrounded by the saliva from von Ebner salivary gland(vEG). Thus, it is important how the secretion of vEG is controlled. This study was designed to elucidate the roles of P2Rs on salivary secretion of vEG. Male Sprague-Dawley rats (about 200 g) were used for this experiment. vEG-rich tissues were obtained from dissecting $500-1,000\;{\mu}m$ thick posterior tongue slices under stereomicroscope view. P2Rs mRNA in vEG acinar cells were identified with RT-PCR. To observe the change in intracellular $Ca^{2+}$ activity, we employed $Ca^{2+}-ion$ specific fluorescence analysis with fura-2. Single acinar cells and cell clusters were isolated by a sequential trypsin/collagenase treatment and were loaded with $10\;{\mu}M$ fura -2 AM for 60 minutes at room temperature. Several agonists and antagonists were used to test a receptor specificity. RT-PCR revealed that the mRNAs of $P2X_4$, $P2Y_1$, $P2Y_2$ and $P2Y_3$ are expressed in vEG acinar cells. The intracellular calcium activity was increased in response to $10\;{\mu}M$ ATP, a P2Rs agonist, and 2-MeSATP, a $P2Y_1$ and $P2Y_2R$ agonist. However, $300\;{\mu}M\;{\alpha}{\beta}-MeATP$, a $P2X_1$ and $P2X_3R$ agonist, did not elicit the response. The responses elicited by $10\;{\mu}M$ ATP and UTP, a $P2Y_2R$ agonists, were maintained when extracellular calcium was removed. $10\;{\mu}M$ suramin, a P2XR antagonist, and reactive blue 2, a P2YR antagonist, partially blocked ATP-induced response. However, when extracellular calciums were removed, suramin did not abolish the responses elicited by ATP. These results suggest that P2Rs play an important role in salivary secretion of vEG acinar cells and the effects of ATP on vEG salivary secretion may be mediated by $P2X_4$, $P2Y_1$, $P2Y_2$, and/or $P2Y_3$.

Efficacy and Safety of Ultrasound-Guided Radiofrequency Ablation for Primary Hyperparathyroidism: A Prospective Study

  • Hui-hui Chai;Yu Zhao;Zeng Zeng;Rui-zhong Ye;Qiao-hong Hu;Hong-feng He;Jung Hwan Baek;Cheng-zhong Peng
    • Korean Journal of Radiology
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    • 제23권5호
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    • pp.555-565
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    • 2022
  • Objective: To assess the efficacy and safety of ultrasound (US)-guided radiofrequency ablation (RFA) in patients with primary hyperparathyroidism (PHPT). Materials and Methods: This prospective study enrolled 39 participants (14 male, 25 female; mean age, 59.5 ± 15.3 [range, 18-87] years) between September 1, 2018, and January 31, 2021. All participants had parathyroid lesions causing PHPT, proven biochemically and through imaging. The imaging features of the PHPT nodules, including the shape, margin, size, composition, and location, were evaluated before treatment. Serum intact parathyroid hormone, calcium, and phosphorus levels; parathyroid nodule volume; and PHPT-related symptoms were recorded before and after treatment. We calculated the technical success, biochemical cure, and clinical cure rates for these patients. Complications were evaluated during and after the ablation. Results: Complete ablation was achieved in 38 of the 39 nodules in the 39 enrolled participants. All the patients were treated in one session. The technical success rate was 97.4% (38/39). The mean follow-up duration was 13.2 ± 4.6 (range, 6.0-24.9) months. At 6 and 12 months post-RFA, the biochemical cure rates were 82.1% (32/39) and 84.4% (27/32), respectively, and the clinical cure rates were 100% (39/39) and 96.9% (31/32), respectively. Only 2.6% (1/39) of the patients had recurrent PHPT. At 1, 3, 6, and 12 months after technically successful RFA, 44.7% (17/38), 34.3% (12/35), 15.8% (6/38), and 12.5% (4/32) of participants, respectively, had elevated eucalcemic parathyroid hormone levels. Recurrent laryngeal nerve paralysis occurred in 5.1% (2/39) of the patients, who recovered spontaneously within 1-3 months. Conclusion: US-guided RFA was effective and safe for PHPT patients. RFA may be an alternative treatment tool for patients who cannot tolerate or refuse to undergo surgery.

Distribution of Taste Receptors in Submandibular and von Ebner Salivary Glands

  • Jun, Yong-Ku;Kim, Se-Nyun;Lee, Cil-Han;Cho, Young-Kyung;Chung, Ki-Myung;Roper, Stephen D.;Kim, Kyung-Nyun
    • International Journal of Oral Biology
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    • 제33권1호
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    • pp.13-23
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    • 2008
  • Taste is a critically important sense for the survival of an organism. However, structure and distribution of taste receptors were only recently investigated. Although expression of the ion channels responsible for the sense of salty taste and acidity was observed in the non-taste cells, receptors for sweet and bitter taste were only identified in taste cells. Salivary glands are involved in the sensing of taste and plays important roles in the transduction of taste. The purpose of this study is to examine whether taste receptors are present in the salivary glands and to provide clues for the investigation of the taste-salivary glands interaction. Using microarray and RT-PCR analyses, the presence of taste receptor mRNAs in the rat von Ebner gland and submandibular gland was confirmed. Type I taste receptors were preferentially expressed in von Ebner gland, whereas type II taste receptors were expressed in both von Ebner gland and submandibular gland. The tastespecific signal tranducing proteins, $G_{\alpha}gustducin$ and phospholipase C ${\beta}2$, were also detected in both salivary glands by immunohistochemistry. Finally, the activation of the calcium signal in response to bitter taste in the acinar cells was also observed. Taken together, these results suggest that taste receptors are present in the von Ebner gland and submandibular gland and that type II taste receptors are functionally active in both salivary glands.

Ginsenoside Rk1 is a novel inhibitor of NMDA receptors in cultured rat hippocampal neurons

  • Ryoo, Nayeon;Rahman, Md. Ataur;Hwang, Hongik;Ko, Sung Kwon;Nah, Seung-Yeol;Kim, Hyoung-Chun;Rhim, Hyewhon
    • Journal of Ginseng Research
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    • 제44권3호
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    • pp.490-495
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    • 2020
  • Background: Ginsenoside Rk1, a saponin component isolated from heat-processed Panax ginseng Meyer, has been implicated in the regulation of antitumor and anti-inflammatory activities. Although our previous studies have demonstrated that ginsenoside Rg3 significantly attenuated the activation of NMDA receptors (NMDARs) in hippocampal neurons, the effects of ginsenosides Rg5 and Rk1, which are derived from heat-mediated dehydration of ginsenoside Rg3, on neuronal NMDARs have not yet been elucidated. Methods: We examined the regulation of NMDARs by ginsenosides Rg5 and Rk1 in cultured rat hippocampal neurons using fura-2-based calcium imaging and whole-cell patch-clamp recordings. Results: The results from our investigation showed that ginsenosides Rg3 and Rg5 inhibited NMDARs with similar potencies. However, ginsenoside Rk1 inhibited NMDARs most effectively among the five compounds (Rg3, Rg5, Rk1, Rg5/Rk1 mixture, and protopanaxadiol) tested in cultured hippocampal neurons. Its inhibition is independent of the NMDA- and glycine-binding sites, and its action seems to involve in an interaction with the polyamine-binding site of the NMDAR channel complex. Conclusion: Taken together, our results suggest that ginsenoside Rk1 might be a novel component contributable to the development of ginseng-based therapeutic treatments for neurodegenerative diseases.

A Case of Severe Hypercalcemia Causing Acute Kidney Injury: An Unusual Presentation of Acute Lymphoblastic Leukemia

  • Hyun, Hye Sun;Park, Peong Gang;Kim, Jae Choon;Hong, Kyun Taek;Kang, Hyoung Jin;Park, Kyung Duk;Shin, Hee Young;Kang, Hee Gyung;Ha, Il Soo;Cheong, Hae Il
    • Childhood Kidney Diseases
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    • 제21권1호
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    • pp.21-25
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    • 2017
  • Severe hypercalcemia is rarely encountered in children, even though serum calcium concentrations above 15-16 mg/dL could be life-threatening. We present a patient having severe hypercalcemia and azotemia. A 14-year-old boy with no significant past medical history was referred to our hospital with hypercalcemia and azotemia. Laboratory and imaging studies excluded hyperparathyroidism and solid tumor. Other laboratory findings including a peripheral blood profile were unremarkable. His hypercalcemia was not improved with massive hydration, diuretics, or even hemodialysis, but noticeably reversed with administration of calcitonin. A bone marrow biopsy performed to rule out the possibility of hematological malignancy revealed acute lymphoblastic leukemia. His hypercalcemia and azotemia resolved shortly after initiation of induction chemotherapy. Results in this patient indicate that a hematological malignancy could present with severe hypercalcemia even though blast cells have not appeared in the peripheral blood. Therefore, extensive evaluation to determine the cause of hypercalcemia is necessary. Additionally, appropriate treatment, viz., hydration or administration of calcitonin is important to prevent complications of severe hypercalcemia, including renal failure and nephrocalcinosis.

Red ginseng extract blocks histamine-dependent itch by inhibition of H1R/TRPV1 pathway in sensory neurons

  • Jang, Yongwoo;Lee, Wook-Joo;Hong, Gyu-Sang;Shim, Won-Sik
    • Journal of Ginseng Research
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    • 제39권3호
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    • pp.257-264
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    • 2015
  • Background: Korean Red Ginseng-a steamed root of Panax ginseng Meyer-has long been used as a traditional medicine in Asian countries. Its antipruritic effect was recently found, but no molecular mechanisms were revealed. Thus, the current study focused on determining the underlying molecular mechanism of Korean Red Ginseng extract (RGE) against histamine-induced itch at the peripheral sensory neuronal level. Methods: To examine the antipruritic effect of RGE, we performed in vivo scratching behavior test in mice, as well as in vitro calcium imaging and whole-cell patch clamp experiments to elucidate underlying molecular mechanisms. Results: The results of our in vivo study confirmed that RGE indeed has an antipruritic effect on histamine-induced scratching in mice. In addition, RGE showed a significant inhibitory effect on histamine-induced responses in primary cultures of mouse dorsal root ganglia, suggesting that RGE has a direct inhibitory effect on sensory neuronal level. Results of further experiments showed that RGE inhibits histamine-induced responses on cells expressing both histamine receptor subtype 1 and TRPV1 ion channel, indicating that RGE blocks the histamine receptor type 1/TRPV1 pathway in sensory neurons, which is responsible for histamine-dependent itch sensation. Conclusion: The current study found for the first time that RGE effectively blocks histamine-induced itch in peripheral sensory neurons. We believe that the current results will provide an insight on itch transmission and will be helpful in understanding how RGE exerts its antipruritic effects.

방사선조사가 구루병 백서의 발치창 치유에 미치는 영향 (Effect of irradiation on wound healing after tooth extraction in the rachitic rats)

  • 김미경;황의환;이상래
    • Imaging Science in Dentistry
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    • 제32권1호
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    • pp.1-10
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    • 2002
  • Purpose: To observe the histopathological changes following irradiation on the wound healing after tooth extraction in the rachitic rats. Materials and Methods: In order to carry out this study, the rats were divided into four groups: Group 1 (normal diet/non-irradiation group), Group 2 (normal diet/irradiation group), Group 3 (rachitogenic diet/non-irradiation group), and Group 4 (rachitogenic diet/irradiation group). Rachitic changes were induced with rachitogenic diet No. 2 (high calcium, low phosphorus, and Vitamin D deficient diet) for 5 weeks. After the extraction of both maxillary first molars of the rats in Group 2 and 4, the head and neck of the rats were irradiated with single absorbed dose of 10 Gy. The rats were sacrificed at the 1st, 5th, 10th, and 15th day after tooth extraction. The specimens including the extraction wound were sectioned, stained with the hematoxylin-eosin and Masson's trichrome method and examined under the light microscope. Results: In the Group 2, the amount of newly formed bone trabeculae on the periphery of extraction socket and osteoblastic activity were reduced. In the Group 3, epithelial fusion was not revealed on the 5th day after toothe extraction and growth rate of osteoid formation was reduced. In the Group 4, necrotized tissue at the outer surface of extraction socket and destructive changes on the alveolar bones were noted on the 10th day. Epithelial fusion was not revealed and large amounts of osteoclast were noted on alveolar bone on the 15th day. Conclusion: The healing process of wound after tooth extraction was retarded by irradiation and especially in the rachitic rats.

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In Vitro and In Vivo Evaluation of Composite Scaffold of BCP, Bioglass and Gelatin for Bone Tissue Engineering

  • Kim, Woo Seok;Nath, Subrata Deb;Bae, Jun Sang;Padalhin, Andrew;Kim, Boram;Song, Myeong Jin;Min, Young Ki
    • 한국재료학회지
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    • 제24권6호
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    • pp.310-318
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    • 2014
  • In this experiment, a highly porous scaffold of biphasic calcium phosphate (BCP) was prepared using the spongereplica method. The BCP scaffold was coated with 58S bioactive glass (BG) and sintered for a second time. The resulting scaffold was coated with gelatin (Gel) and cross-linked with [3-(3-dimethyl aminopropyl) carbodiimide] and N-Hydroxysuccinamide (EDC-NHS). The initial average pore size of the scaffold ranged from 300 to $700{\mu}m$, with more than 85 % porosity. The coating of BG and Gel had a significant effect on the scaffold-pore size, decreasing scaffold porosity while increasing mechanical strength. The material and surface properties were evaluated by means of several experiments involving scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) and X-ray diffraction (XRD). Cytotoxicity was evaluated using MTT assay and confocal imaging of MC3T3-E1 pre-osteoblast cells cultured in vitro. Three types of scaffold (BCP, BCP-BG and BCP-BG-Gel) were implanted in a rat skull for in vivo evaluation. After 8 weeks of implantation, bone regeneration occurred in all three types of sample. Interestingly, regeneration was found to be greater (geometrically and physiologically) for neat BCP scaffolds than for two other kinds of composite scaffolds. However, the other two types of scaffolds were still better than the control (i.e., defect without treatment).