• Biomolecules & Therapeutics
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• 제26권5호
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• pp.439-445
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• 2018

T-type calcium channels are low voltage-activated calcium channels that evoke small and transient calcium currents. Recently, T-type calcium channels have been implicated in neurodevelopmental disorders such as autism spectrum disorder and neural tube defects. However, their function during embryonic development is largely unknown. Here, we investigated the function and expression of T-type calcium channels in embryonic neural progenitor cells (NPCs). First, we compared the expression of T-type calcium channel subtypes (CaV3.1, 3.2, and 3.3) in NPCs and differentiated neural cells (neurons and astrocytes). We detected all subtypes in neurons but not in astrocytes. In NPCs, CaV3.1 was the dominant subtype, whereas CaV3.2 was weakly expressed, and CaV3.3 was not detected. Next, we determined CaV3.1 expression levels in the cortex during early brain development. Expression levels of CaV3.1 in the embryonic period were transiently decreased during the perinatal period and increased at postnatal day 11. We then pharmacologically blocked T-type calcium channels to determine the effects in neuronal cells. The blockade of T-type calcium channels reduced cell viability, and induced apoptotic cell death in NPCs but not in differentiated astrocytes. Furthermore, blocking T-type calcium channels rapidly reduced AKT-phosphorylation (Ser473) and $GSK3{\beta}$-phosphorylation (Ser9). Our results suggest that T-type calcium channels play essential roles in maintaining NPC viability, and T-type calcium channel blockers are toxic to embryonic neural cells, and may potentially be responsible for neurodevelopmental disorders.

• The Korean Journal of Physiology and Pharmacology
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• 제4권6호
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• pp.427-437
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• 2000

The early studies of cardiac and smooth muscle cells provided evidence for two different calcium channels, the L-type (also called high-voltage activated [HVA]) and T-type (low-voltage activated [LVA]). These calcium channels provided calcium for muscle contractions and pace-making activities. As might be expected, the number of different calcium channels increased when researchers studied neurons and the identification of the neuronal calcium channels has proven to be much more difficult than with the muscle calcium channels. There are two reasons for this difficulty; (1) a larger number of different calcium channels in neurons and (2) many of the different calcium channels have similar kinetic properties. This review uses the N-type calcium channel to illustrate the difficulties in identifying and characterizing calcium channels in neurons. It shows that the discovery of toxins that can specifically block single calcium channel types has made it possible to easily and rapidly discern the physiological roles of the different calcium channels in the neuron, Without these toxins it is unlikely that progress would have been as rapid.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2002년도 제9회 학술 발표회 프로그램과 논문초록
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• pp.20-21
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• 2002

The inflow of Ca$\^$2＋/ through voltage-activated T-type calcium channels (T-channels) regulates a variety of cellular functions including neuronal excitability, cardiac pacemaker activity, hormone secretion, smooth muscle contraction, and fertilization. Not only are T-channels enormously important for the normal operation of cells, they also playa critical role in pathophysiological conditions such as cardiac hypertrophy and absence epilepsy.(omitted)

• Biomolecules & Therapeutics
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• 제28권5호
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• pp.389-396
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• 2020

Valproic acid is a clinically used mood stabilizer and antiepileptic drug. Valproic acid has been suggested as a teratogen associated with the manifestation of neurodevelopmental disorders, such as fetal valproate syndrome and autism spectrum disorders, when taken during specific time window of pregnancy. Previous studies proposed that prenatal exposure to valproic acid induces abnormal proliferation and differentiation of neural progenitor cells, presumably by inhibiting histone deacetylase and releasing the condensed chromatin structure. Here, we found valproic acid up-regulates the transcription of T-type calcium channels by inhibiting histone deacetylase in neural progenitor cells. The pharmacological blockade of T-type calcium channels prevented the increased proliferation of neural progenitor cells induced by valproic acid. Differentiated neural cells from neural progenitor cells treated with valproic acid displayed increased levels of calcium influx in response to potassium chloride-induced depolarization. These results suggest that prenatal exposure to valproic acid up-regulates T-type calcium channels, which may contribute to increased proliferation of neural progenitor cells by inducing an abnormal calcium response and underlie the pathogenesis of neurodevelopmental disorders.

• Molecules and Cells
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• 제22권1호
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• pp.51-57
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• 2006

Haloperidol is a classical neuroleptic drug that is still in clinical use and can lead to abnormal motor activity following repeated administration. However, there is little knowledge of how it triggers neuronal impairment. In this study, we report that it induced calcium ion influx via L-type calcium channels and that the elevation of calcium ions induced by haloperidol appeared to render hippocampal cells more susceptible to oxidative stress. Indeed, the level of cytotoxic reactive oxygen species (ROS) and the expression of pro-apoptotic Bax increased in response to oxidative stress in haloperidol-treated cells, and these effects were inhibited by verapamil, a specific L-type calcium channel blocker, but not by the T-type calcium channel blocker, mibefradil. These findings indicate that haloperidol induces calcium ion influx via L-type calcium channels and that this calcium influx influences neuronal fate.

• The Korean Journal of Physiology and Pharmacology
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• 제13권3호
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• pp.241-249
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• 2009

Although extracellular $Ca^{2+}$ entry through the voltage-dependent $Ca^{2+}$ channels plays an important role in the spontaneous phasic contractions of the pregnant rat myometrium, the role of the T-type $Ca^{2+}$ channels has yet to be fully identified. The aim of this study was to investigate the role of the T-type $Ca^{2+}$ channel in the spontaneous phasic contractions of the rat myometrium. Spontaneous phasic contractions and $[Ca^{2+}]_i$ were measured simultaneously in the longitudinal strips of female Sprague-Dawley rats late in their pregnancy (on day 18 ${\sim}$ 20 of gestation: term=22 days). The expression of T-type $Ca^{2+}$ channel mRNAs or protein levels was measured. Cumulative addition of low concentrations (< 1 ${\mu}M$) of nifedipine, a L-type $Ca^{2+}$ channel blocker, produced a decrease in the amplitude of the spontaneous $Ca^{2+}$ transients and contractions with no significant change in frequency. The mRNAs and proteins encoding two subunits (${\alpha}$ 1G, ${\alpha}$ 1H) of the T-type $Ca^{2+}$ channels were expressed in longitudinal muscle layer of rat myometrium. Cumulative addition of mibefradil, NNC 55-0396 or nickel induced a concentration-dependent inhibition of the amplitude and frequency of the spontaneous $Ca^{2+}$ transients and contractions. Mibefradil, NNC 55-0396 or nickel also attenuated the slope of rising phase of spontaneous $Ca^{2+}$ transients consistent with the reduction of the frequency. It is concluded that T-type $Ca^{2+}$ channels are expressed in the pregnant rat myometrium and may play a key role for the regulation of the frequency of spontaneous phasic contractions.

• 생명과학회지
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• 제32권2호
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• pp.108-118
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• 2022

뇌실하 영역은 뇌에서 신경줄기세포가 분포하는 곳으로 평생에 걸쳐 새로운 신경세포를 생성하는 곳이다. 많은 세포 안팎의 인자들이 신경줄기세포의 세포 증식과 신경세포로의 분화에 영향을 미친다. 최근 들어, L-type 칼슘 채널이 신경계의 발달을 조절하고 뇌실하 영역에 있는 신경줄기세포, 신경세포로 분화 중인 세포, 그리고 성숙한 신경세포에 분포한다고 밝혀졌다. L-type 칼슘 채널의 저해제인 nifedipine은 고혈압의 치료제로 오랜 기간 사용되어 왔다. 신경줄기세포에 nifedipine을 사용하여 L-type 칼슘 채널을 저해하는 연구는 많이 없는 상황이다. 이번 연구에서, 우리는 5일령 쥐의 뇌실하 영역에서 배양한 신경줄기세포에 nifedipine을 처리하여 신경세포로의 분화에 미치는 영향을 관찰하였다. Nifedipine은 Tuj1을 발현하는 신경세포의 수를 증가시킨 반면, Olig2를 발현하는 희소 돌기 아교 세포(oligodendrocytes)의 수에는 큰 영향을 미치지 않았다. Nifedipine은 S기를 표지하는 5-ethynyl-2'-deoxyuridine (EdU)가 들어간 세포의 수를 증가시켰고, 세포 분열시 나타나는 인산화된 히스톤 H3(PH3)를 발현하는 세포의 수를 증가시켰다. Nifedipine은 신경세포로의 분화를 촉진하는 Dlx2 유전자의 전사를 증가시켰고, 초기 신경세포에서 보이는 Mash1의 양도 증가시켰다. Nifedipine 외 또다른 L-type 칼슘 채널의 저해제인 verapamil을 처리하자, 신경세포로의 분화가 소폭 증가하였으나, 통계적 유의미성은 매우 낮았다. T-type 칼슘 채널의 저해제 유전자인 Cav3.1, Cav3.2, Cav3.3가 발현함을 관찰하여, T-type 칼슘 채널의 저해제인 pimozide를 신경줄기세포에 처리하였으나, 신경세포로의 분화에는 변화가 없었다. 이러한 결과를 통해 nifedipine이 신경줄기세포의 초기 분화를 증진함을 알 수 있으며, L-type 칼슘 채널이 신경세포로의 분화에 관여함을 알 수 있다.

• 한국발생생물학회지:발생과생식
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• 제24권4호
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• pp.297-306
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• 2020

Repetitive changes in the intracellular calcium concentration ([Ca²⁺]i) triggers egg activation, including cortical granule exocytosis, resumption of second meiosis, block to polyspermy, and initiating embryonic development. [Ca²⁺]i oscillations that continue for several hours, are required for the early events of egg activation and possibly connected to further development to the blastocyst stage. The sources of Ca²⁺ ion elevation during [Ca²⁺]i oscillations are Ca²⁺ release from endoplasmic reticulum through inositol 1,4,5 tri-phosphate receptor and Ca²⁺ ion influx through Ca²⁺ channel on the plasma membrane. Ca²⁺ channels have been characterized into voltage-dependent Ca²⁺ channels (VDCCs), ligand-gated Ca²⁺ channel, and leak-channel. VDCCs expressed on muscle cell or neuron is specified into L, T, N, P, Q, and R type VDCs by their activation threshold or their sensitivity to peptide toxins isolated from cone snails and spiders. The present study was aimed to investigate the localization pattern of N and P/Q type voltage-dependent calcium channels in mouse eggs and the role in fertilization. [Ca²⁺]i oscillation was observed in a Ca²⁺ contained medium with sperm factor or adenophostin A injection but disappeared in Ca²⁺ free medium. Ca²⁺ influx was decreased by Lat A. N-VDCC specific inhibitor, ω-Conotoxin CVIIA induced abnormal [Ca²⁺]i oscillation profiles in SrCl₂ treatment. N or P/Q type VDC were distributed on the plasma membrane in cortical cluster form, not in the cytoplasm. Ca²⁺ influx is essential for [Ca²⁺]i oscillation during mammalian fertilization. This Ca²⁺ influx might be controlled through the N or P/Q type VDCCs. Abnormal VDCCs expression of eggs could be tested in fertilization failure or low fertilization eggs in subfertility women.

• Molecules and Cells
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• 제25권2호
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• pp.242-246
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• 2008

To assess the role of $\alpha_{1G}$ T-type $Ca^{2+}$ channels in neuropathic pain after L5 spinal nerve ligation, we examined behavioral pain susceptibility in mice lacking $Ca_{V}3.1$ (${\alpha}_{1G}{^{-/-}}$), the gene encoding the pore-forming units of these channels. Reduced spontaneous pain responses and an increased threshold for paw withdrawal in response to mechanical stimulation were observed in these mice. The ${{\alpha}_{1G}}^{-/-}$ mice also showed attenuated thermal hyperalgesia in response to both low-(IR30) and high-intensity (IR60) infrared stimulation. Our results reveal the importance of ${\alpha}_{1G}$ T-type $Ca^{2+}$ channels in the development of neuropathic pain, and suggest that selective modulation of ${\alpha}_{1G}$ subtype channels may provide a novel approach to the treatment of allodynia and hyperalgesia.

• Clinical and Experimental Reproductive Medicine
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• 제28권1호
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• pp.13-24
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• 2001

Objective: In muscle and neuronal cells, calcium channels have been classified by electrophysiological and pharmacological properties into (1) voltage-dependent $Ca^{2+}$-channel (1) P/Q-type $Ca^{2+}$-channel (2) N-type $Ca^{2+}$-channel (3) L-type $Ca^{2+}$-channel (4) T-type $Ca^{2+}$-channel (5) R-type $Ca^{2+}$-channel. The present study was done in order to investigate whether there is any difference in $Ca^{2+}$-channel distribution between activated and normally fertilized embryos. Methods: The immunocytochemical method was used to identify the existence of voltage-dependent $Ca^{2+}$-channels in parthenogenetically activated 2-cell embryos by ethanol and $SrCl_2$ treatment. These 2-cell embryos were obtained by exposure to 6% ethanol for 6 min and to 10 mM $SrCl_2$ for 2h. Results: P/Q-type $Ca^{2+}$-channels and L-type $Ca^{2+}$-channels have been identified. Whereas, three type of $Ca^{2+}$-channel P/Q-type, N-type, L-type have been identified in 2-cell embryos fertilized in vivo. Conclusion: Activation by ethanol was faster than those by $SrCl_2$. However, there was difference in DAB staining of the embryos between ethanol and $SrCl_2$ treatment (87.7% and 54.1 %). Intensity of staining was also different between ethanol- and $SrCl_2$-treated group. However, it has not been known why there was some difference in DAB staining and staining intensity in the present study.