• 제목/요약/키워드: Extracellular recording

검색결과 34건 처리시간 0.018초

Background $K^+$ channel currents in WEHI-231 cells, immature B lymphocytes

  • Nam, Joo-Hyun;Woo, Ji-Eun;Kim, Tae-Jin;Uhm, Dae-Yong;Kim, Sung-Joon
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.39-39
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    • 2003
  • In our previous study, WEHI-231, an immature B cell line, showed intractable increase in [C $a^{2+}$]$_{c}$ after the B-cell receptor (BCR) ligation and treatment with 2-aminoethoxydiphenylborate (2-APB), which was never observed in Bal-17, a mature B cell line (Nam et al., 2003, FEBS Lett). In this study, a whole cell voltage clamp study revealed a specific expression of a novel type of $K^{+}$ current, namely voltage-independent background-type $K^{+}$ channels (IK-bg), in WEHI-231 cells. IK-bg was dramatically increase by the application of 2-APB (50 $\square$M), which induced severe hyperpolarization of WEHI-231 from -45 ㎷ to -90 ㎷, When dialyzed with $Mg^{2+}$ and ATP-free pipette solution, a spontaneous development of IK-bg and membrane hyperpolarization were observed. IK-bg was insensitive to classical $K^{+}$ channel blockers (TEA, glibenclamide, $Ba^{2+}$(1 mM)), whereas blocked by quinine and quinidine in a voltage-dependent manner ($IC_{50}$/=6~9 $\square$M at +60㎷). Phorbol myrstate, a PKC activator, decreased the amplitude of IK-bg. Extracellular acidification (pH 6.5) slightly inhibited IK-bg. Arachidonic acid, riluzole, or hyposmotic stress could not affect the IK-bg after the full development by the intracellular dialysis with Mg-ATP-free solution. In a cell-attached mode of single channel recording from WEHI231, we found two types of voltage-independent $K^{+}$ channels with unitary conductance of 300 pS and 120 pS, respectively. Both channels showed very short mean open times and their open probabilities were increase by the application of 2-APB. In Bal-17 cells, no such $K^{+}$ current was observed in 50 cells tested. In summary, WEHI-231 immature B cells express background $K^{+}$ channels. The pharmacological properties and the large unitary conductance suggest that novel types of two-pore domain $K^{+}$ channels (2-P-K channels) might be expressed in WEHI-231, which may provide an intriguing targets of signal transduction in the immature B lymphocytes.e B lymphocytes.

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흰쥐 안면신경핵 세포의 전기생리학적 및 형태학적 특성 (ELECTROPHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF FACIAL NUCLEUS IN RAT)

  • 최병주;조진화;배용철;김영진
    • 대한소아치과학회지
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    • 제27권3호
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    • pp.400-409
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    • 2000
  • 흰쥐의 안면신경핵을 구성하는 신경세포들의 시냅스 연결 양태 및 세포막 특성을 규명하기 위해 in vivo 필드전위 및 세포 내 전위 측정법을 이용하여 전기생리적 반응을 관찰하였다. 말초 안면신경 분지를 역행성으로 전기자극시 자극세기에 비례하여 전위의 크기가 증가되었고 필드 전위의 양태는 두 가지 반응으로 나타났는데 전기자극 직후 1ms 부근에서 정점을 나타내는 양태와 이와 더불어 $7\sim8ms$ 부근에서 후기 정점을 동반하는 양태가 있었다. 안면신경핵은 염색시 내측, 배외측, 중간측 및 외측등 4부분의 소핵으로 구분되었다. Neurobiotin으로 채워진 단일 신경세포를 형태학적으로 재구축하였는데 세포체는 추체형태를 나타내었고 주 수상돌기는 모든 방향으로 뻗어져 있었고 각 수상돌기의 영역은 해당 소핵 내에 한정되어 있었다. 일련의 과분극 전류 $(-1.2\sim+1.2nA)$를 세포내에 가하였을 때 동반되는 세포내 전위변화를 입력저항 값으로 계산하였을 때 그 기울기가 직선형으로 나타났다. 탈분극 전류를 세포내 주입시 지속적인 활동성 전위가 나타났으며 전류의 크기에 비례하여 각 전위의 개수가 증가하였고 spike-빈도 적응 현상이 나타났다. 그러나 시간 의존성 내향성 정류현상은 관찰되지 않았고 anodal break excitation이 나타났다. 이상의 실험결과로 보아 안면신경핵을 구성하고 있는 세포들 사이의 시냅스는 다양한 형태로 존재할 가능성이 있다고 사료되며 이들 시냅스간의 변화를 통하여 안면 신경마비, 반쪽 안면 경련, hypoglossal-facial anastomosis등에서 나타날 수 있는 임상적 신경성 증상 기전을 설명할 수 있을 것으로 여겨진다.

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Single-Channel Recording of TASK-3-like $K^+$ Channel and Up-Regulation of TASK-3 mRNA Expression after Spinal Cord Injury in Rat Dorsal Root Ganglion Neurons

  • Jang, In-Seok;La, Jun-Ho;Kim, Gyu-Tae;Lee, Jeong-Soon;Kim, Eun-Jin;Lee, Eun-Shin;Kim, Su-Jeong;Seo, Jeong-Min;Ahn, Sang-Ho;Park, Jae-Yong;Hong, Seong-Geun;Kang, Da-Won;Han, Jae-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • 제12권5호
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    • pp.245-251
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    • 2008
  • Single-channel recordings of TASK-1 and TASK-3, members of two-pore domain $K^+$ channel family, have not yet been reported in dorsal root ganglion (DRG) neurons, even though their mRNA and activity in whole-cell currents have been detected in these neurons. Here, we report single-channel kinetics of the TASK-3-like $K^+$ channel in DRG neurons and up-regulation of TASK-3 mRNA expression in tissues isolated from animals with spinal cord injury (SCI). In DRG neurons, the single-channel conductance of TASK-3-like $K^+$ channel was $33.0{\pm}0.1$ pS at - 60 mV, and TASK-3 activity fell by $65{\pm}5%$ when the extracellular pH was changed from 7.3 to 6.3, indicating that the DRG $K^+$ channel is similar to cloned TASK-3 channel. TASK-3 mRNA and protein levels in brain, spinal cord, and DRG were significantly higher in injured animals than in sham-operated ones. These results indicate that TASK-3 channels are expressed and functional in DRG neurons and the expression level is up-regulated following SCI, and suggest that TASK-3 channel could act as a potential background $K^+$ channel under SCI-induced acidic condition.

마그네슘이온이 적출한 기니피그 대동맥평활근과 흰쥐 자궁평활근의 수축성에 미치는 효과에 관한 연구 (Effect of Magnesium on the Contractility of the Isolated Guinea-Pig Aortic and Rat Smooth Muscles)

  • 안혁;황상익
    • Journal of Chest Surgery
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    • 제23권3호
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    • pp.452-464
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    • 1990
  • It is well known that extracellular Calcium plays a very important role in several steps of smooth muscle excitability and contractility, and there have been many concerns about factors influencing the distribution of extracellular Ca++ and the Ca++ flux through the cell membrane of the smooth muscle. Based on the assumption that Mg++ may also play an important role in the excitation and contraction processes of the smooth muscle by taking part in affecting Ca++ distribution and flux, many researches are being performed about the exact role of Mg++, especially in the vascular smooth muscle. But yet the effect of Mg++ in the smooth muscle activity is not clarified, and moreover the mechanism of Mg++ action is almost completely unknown. Present study attempted to clarify the effect of Mg++ on the excitability and contractility in the multiunit and unitary smooth muscle, and the mechanism concerned in it. The preparations used were the guinea-pig aortic strip as the experimental material of the multiunit smooth muscle and the rat uterine strip as the one of the unitary smooth muscle. The tissues were isolated from the sacrificed animal and were prepared for recording the isometric contraction. The effects of Mg++ and Ca++ were examined on the electrically driven or spontaneous contraction of the preparations. And the effects of these ions were also studied on the K+ or norepinephrine contracture. All experiments were performed in tris-buffered Tyrode solution which was aerated with 100% 02 and kept at 35oC. The results obtained were as follows: 1] Mg++ suppressed the phasic contraction induced by electrical field stimulation dose-dependently in the guinea-pig aortic strip, while the high concentration of Ca++ never recovered the decreased tension. These phenomena were not changed by the a - or b - adrenergic blocker. 2]Mg++ played the suppressing effect on the low concentration [20 and 40 mM] of K+-contracture in the aortic muscle, but the effect was not shown in the case of 100mM K+-contracture. 3] Mg++ also suppressed the contracture induced by norepinephrine in the aortic preparation. And the effect of Mg++ was most prominent in the contracture by the lowest [10 mM] concentration of norepinephrine. 4] In both the spontaneous and electrically driven contractions of the uterine strip, Mg++ decreased the amplitude of peak tension, and by the high concentration of Ca++ the amplitude of tension was recovered unlike the aortic muscle. 5] The frequency of the uterine spontaneous contraction increased as the [Ca++] / [Mg++] ratio increased up to 2, but the frequency decreased above this level. 6] Mg++ decreased the tension of the low[20 and 40mM] K+-contracture in the uterine smooth muscle, but the effect did not appear in the 100mM K+-contracture. From the above results, the following conclusion could be made. 1] Mg++ seems to suppress the contractility directly by acting on the smooth muscle itself, besides through the indirect action on the nerve terminal, in both the aortic and uterine smooth muscles. 2] The fact that the depressant effect of Mg++ on the K+-contracture is in inverse proportion to an increase of K+ concentration appears resulted from the extent of the opening state of the Ca++ channel. 3] Mg++ may play a depressant role on both the potential dependent and the receptor-operated Ca++ channels. 4] The relationship between the actions of Mg++ and Ca++ seems to be competitive in uterine muscle and non-competitive in aortic strip.

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