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Article ; Online: Analyses of the Mode of Action of an Alpha-Adrenoceptor Blocker in Substantia Gelatinosa Neurons in Rats.

Uta, Daisuke / Hattori, Tsuyoshi / Yoshimura, Megumu

International journal of molecular sciences

2021 Volume 22, Issue 17

Abstract: To elucidate why naftopidil increases the frequency of spontaneous synaptic currents in only some substantia gelatinosa (SG) neurons, post-hoc analyses were performed. Blind patch-clamp recording was performed using slice preparations of SG neurons from ... ...

Abstract	To elucidate why naftopidil increases the frequency of spontaneous synaptic currents in only some substantia gelatinosa (SG) neurons, post-hoc analyses were performed. Blind patch-clamp recording was performed using slice preparations of SG neurons from the spinal cords of adult rats. Spontaneous inhibitory and excitatory postsynaptic currents (sIPSCs and sEPSCs, respectively) were recorded. The ratios of the frequency and amplitude of the sIPSCs and sEPSCs following the introduction of naftopidil compared with baseline, and after the application of naftopidil, serotonin (5-HT), and prazosin, compared with noradrenaline (NA) were evaluated. First, the sIPSC analysis indicated that SG neurons reached their full response ratio for NA at 50 μM. Second, they responded to 5-HT (50 μM) with a response ratio similar to that for NA, but prazosin (10 μM) did not change the sEPSCs and sIPSCs. Third, the highest concentration of naftopidil (100 μM) led to two types of response in the SG neurons, which corresponded with the reactions to 5-HT and prazosin. These results indicate that not all neurons were necessarily activated by naftopidil, and that the micturition reflex may be regulated in a sophisticated manner by inhibitory mechanisms in these interneurons.
MeSH term(s)	Adrenergic alpha-Antagonists/pharmacology ; Animals ; Excitatory Postsynaptic Potentials/drug effects ; Excitatory Postsynaptic Potentials/physiology ; Inhibitory Postsynaptic Potentials/drug effects ; Inhibitory Postsynaptic Potentials/physiology ; Male ; Membrane Potentials/drug effects ; Membrane Potentials/physiology ; Naphthalenes/pharmacology ; Neurons/drug effects ; Neurons/physiology ; Norepinephrine/pharmacology ; Patch-Clamp Techniques/methods ; Piperazines/pharmacology ; Prazosin/pharmacology ; Rats, Sprague-Dawley ; Serotonin/pharmacology ; Substantia Gelatinosa/cytology ; Substantia Gelatinosa/drug effects ; Substantia Gelatinosa/physiology ; Synaptic Transmission/drug effects ; Synaptic Transmission/physiology ; Rats
Chemical Substances	Adrenergic alpha-Antagonists ; Naphthalenes ; Piperazines ; Serotonin (333DO1RDJY) ; naftopidil (R9PHW59SFN) ; Norepinephrine (X4W3ENH1CV) ; Prazosin (XM03YJ541D)
Language	English
Publishing date	2021-09-06
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms22179636
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Article ; Online: Analyses of the Mode of Action of an Alpha-Adrenoceptor Blocker in Substantia Gelatinosa Neurons in Rats

Daisuke Uta / Tsuyoshi Hattori / Megumu Yoshimura

International Journal of Molecular Sciences, Vol 22, Iss 9636, p

2021 Volume 9636

Abstract	To elucidate why naftopidil increases the frequency of spontaneous synaptic currents in only some substantia gelatinosa (SG) neurons, post-hoc analyses were performed. Blind patch-clamp recording was performed using slice preparations of SG neurons from the spinal cords of adult rats. Spontaneous inhibitory and excitatory postsynaptic currents (sIPSCs and sEPSCs, respectively) were recorded. The ratios of the frequency and amplitude of the sIPSCs and sEPSCs following the introduction of naftopidil compared with baseline, and after the application of naftopidil, serotonin (5-HT), and prazosin, compared with noradrenaline (NA) were evaluated. First, the sIPSC analysis indicated that SG neurons reached their full response ratio for NA at 50 μM. Second, they responded to 5-HT (50 μM) with a response ratio similar to that for NA, but prazosin (10 μM) did not change the sEPSCs and sIPSCs. Third, the highest concentration of naftopidil (100 μM) led to two types of response in the SG neurons, which corresponded with the reactions to 5-HT and prazosin. These results indicate that not all neurons were necessarily activated by naftopidil, and that the micturition reflex may be regulated in a sophisticated manner by inhibitory mechanisms in these interneurons.
Keywords	substantia gelatinosa ; α1-adrenoceptor ; naftopidil ; noradrenaline ; 5-HT ; blind patch-clamp recording ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	590
Language	English
Publishing date	2021-09-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: In vivo electrophysiological analysis of mechanisms of monoaminergic pain inhibitory systems.

Yoshimura, Megumu / Furue, Hidemasa

Pain

2017 Volume 158 Suppl 1, Page(s) S85–S91

MeSH term(s)	Animals ; Biogenic Monoamines/metabolism ; Biogenic Monoamines/pharmacology ; Electrophysiology/methods ; Humans ; Pain/metabolism ; Pain/pathology ; Pain/physiopathology ; Sensory Receptor Cells/drug effects ; Sensory Receptor Cells/physiology
Chemical Substances	Biogenic Monoamines
Language	English
Publishing date	2017-02-24
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	193153-2
ISSN	1872-6623 ; 0304-3959
ISSN (online)	1872-6623
ISSN	0304-3959
DOI	10.1097/j.pain.0000000000000844
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Article: Effect of Alpha 1-Adrnoceptor Antagonists on Postsynaptic Sensitivity in Substantia Gelatinosa Neurons From Lumbosacral Spinal Cord in Rats Using Slice Patch-Clamp Technique for mEPSC.

Uta, Daisuke / Hattori, Tsuyoshi / Yoshimura, Megumu

International neurourology journal

2020 Volume 24, Issue 2, Page(s) 135–143

Abstract: Purpose: Alpha1-adrenoceptors participate in improving storage symptoms of male lower urinary tract symptoms (LUTS). However, the mechanism of action of these compounds remains unclear. To clarify the mechanism of the α1-adrenoceptor antagonists, the ... ...

Abstract	Purpose: Alpha1-adrenoceptors participate in improving storage symptoms of male lower urinary tract symptoms (LUTS). However, the mechanism of action of these compounds remains unclear. To clarify the mechanism of the α1-adrenoceptor antagonists, the amplitude of miniature excitatory postsynaptic currents (mEPSCs) was analyzed in the lumbosacral spinal cord in rats. Methods: Male adult Sprague-Dawley rats were used. Blind whole-cell patch-clamp recordings were performed on substantia gelatinosa (SG) neurons in spinal cord slice preparations. The amplitude of mEPSCs was recorded in individual SG neurons to which α1-adrenoceptors (100μM naftopidil, 100μM tamsulosin, and 30μM silodosin) were applied sequentially with intervening washout periods. Individual amplitudes were analyzed. Results: Pearson correlation coefficients (r) for the amplitudes of mEPSCs between the baseline and postadministration of α1- adrenoceptor antagonists indicated changes of the amplitude ranked in the order of naftopidil (r =0.393), tamsulosin (r=0.738), and silodosin (r=0.944). Together, the α1-adrenoceptor antagonists yielded significant increases in the amplitude of mEPSCs in SG neurons (n=108, P=0.012). However, the effects of each α1-adrenoceptor antagonist on the amplitude were as follows (relative to the baseline; n=36 each): naftopidil, P=0.129; tamsulosin, P=0.201; and silodosin, P=0.005. The rate of response to naftopidil for the outward current was relatively high among the α1-adrenoceptor blockers. An inward current was observed only with the naftopidil application. Conclusion: Alpha1-adrenoceptor antagonists changed the amplitudes of mEPSCs in a subset of SG neurons in slices prepared from the L6-S1 levels of rat spine. Although the α1-adrenoceptor antagonists generated inward or outward currents in the SG neurons, different rates of response were observed with each antagonist. These results are important for understanding the mechanisms of action (at the spinal level) of α1-adrenoceptor antagonists for the storage symptoms of male LUTS.
Language	English
Publishing date	2020-06-30
Publishing country	Korea (South)
Document type	Journal Article
ZDB-ID	2584447-7
ISSN	2093-6931 ; 2093-4777
ISSN (online)	2093-6931
ISSN	2093-4777
DOI	10.5213/inj.1938250.125
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Article: Differential Effects of Alpha 1-Adrenoceptor Antagonists on the Postsynaptic Sensitivity: Using Slice Patch-Clamp Technique for Inhibitory Postsynaptic Current in Substantia Gelatinosa Neurons From Lumbosacral Spinal Cord in Rats.

Uta, Daisuke / Hattori, Tsuyoshi / Yoshimura, Megumu

International neurourology journal

2020 Volume 24, Issue 2, Page(s) 127–134

Abstract: Purpose: Alpha1-adrenoceptors participate in improving storage symptoms of male lower urinary tract symptoms. However, the mechanism of action of these compounds remains unclear. The goal of the present study was to clarify the effect of α1- ... ...

Abstract	Purpose: Alpha1-adrenoceptors participate in improving storage symptoms of male lower urinary tract symptoms. However, the mechanism of action of these compounds remains unclear. The goal of the present study was to clarify the effect of α1- adrenoceptor antagonists on γ-aminobutyric acid (GABA)/glycine-mediated outward currents of the inhibitory postsynaptic current (IPSC) in substantia gelatinosa (SG) neurons from the lumbosacral spinal cord in rats. Methods: Male adult Sprague-Dawley rats were used. Blind whole-cell patch-clamp recordings were performed in SG neurons from isolated spinal cord slice preparations. IPSCs were recorded in individual SG neurons to which naftopidil (100μM), tamsulosin (100μM), silodosin (30μM), or prazosin (10μM) were applied sequentially with intervening washout periods. Strychnine (2μM), bicuculline (10μM), or tetrodotoxin (TTX)(1μM) were added before naftopidil. Individual outward currents were analyzed. Results: The bath application of naftopidil, yielded outward IPSCs in 13 of 52 SG neurons. The naftopidil response was unchanged in the presence of TTX. Regression analysis of the outward currents between the 1st and 2nd applications of naftopidil revealed a Pearson correlation coefficient of 0.996 with a line slope of 0.983. The naftopidil-induced outward current was attenuated in the presence of strychnine and/or bicuculline. The GABA/glycine-mediated outward currents induced by tamsulosin, silodosin, and prazosin were smaller than those obtained with naftopidil. Conclusion: Naftopidil-induced GABA/glycine-mediated outward currents in a subset of SG neurons prepared from the L6- S1 level of rat spinal cord. The results indicated that α1-adrenoceptor antagonists, particularly naftopidil, induce neural suppression (in part) by mediating hyperpolarization. The response is associated with glycinergic and/or GABAergic neural transmission. Naftopidil may suppress the micturition reflex and improve urinary storage symptoms as a subsidiary effect resulting from hyperpolarization in SG neurons of the spinal cord.
Language	English
Publishing date	2020-06-30
Publishing country	Korea (South)
Document type	Journal Article
ZDB-ID	2584447-7
ISSN	2093-6931 ; 2093-4777
ISSN (online)	2093-6931
ISSN	2093-4777
DOI	10.5213/inj.1938248.124
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Article ; Online: L-bupivacaine Inhibition of Nociceptive Transmission in Rat Peripheral and Dorsal Horn Neurons.

Uta, Daisuke / Koga, Kohei / Furue, Hidemasa / Imoto, Keiji / Yoshimura, Megumu

Anesthesiology

2020 Volume 134, Issue 1, Page(s) 88–102

MeSH term(s)	Anesthetics, Local/pharmacology ; Animals ; Bupivacaine/pharmacology ; Excitatory Postsynaptic Potentials/drug effects ; Male ; Nerve Fibers, Myelinated/drug effects ; Nerve Fibers, Unmyelinated/drug effects ; Neurons/drug effects ; Nociception/drug effects ; Patch-Clamp Techniques ; Peripheral Nerves/drug effects ; Posterior Horn Cells/drug effects ; Rats ; Rats, Sprague-Dawley ; Sodium Channels/drug effects ; Stereoisomerism ; Synaptic Transmission/drug effects ; Tetrodotoxin/pharmacology
Chemical Substances	Anesthetics, Local ; Sodium Channels ; Tetrodotoxin (4368-28-9) ; Bupivacaine (Y8335394RO)
Language	English
Publishing date	2020-11-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	269-0
ISSN	1528-1175 ; 0003-3022
ISSN (online)	1528-1175
ISSN	0003-3022
DOI	10.1097/ALN.0000000000003596
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Article ; Online: Mechanisms of the analgesic effect of calcitonin on chronic pain by alteration of receptor or channel expression.

Ito, Akitoshi / Yoshimura, Megumu

Molecular pain

2017 Volume 13, Page(s) 1744806917720316

Abstract: The polypeptide hormone calcitonin is well known clinically for its ability to relieve osteoporotic back pain and neuropathic pain such as spinal canal stenosis, diabetic neuropathy, chemotherapy-induced neuropathy, and complex regional pain syndrome. ... ...

Abstract	The polypeptide hormone calcitonin is well known clinically for its ability to relieve osteoporotic back pain and neuropathic pain such as spinal canal stenosis, diabetic neuropathy, chemotherapy-induced neuropathy, and complex regional pain syndrome. Because the analgesic effects of calcitonin have a broad range, the underlying mechanisms of pain relief by calcitonin are largely unknown. However, recent studies using several types of chronic pain models combined with various methods have been gradually clarifying the mechanism. Here, we review the mechanisms of the analgesic action of calcitonin on ovariectomy-induced osteoporotic and neuropathic pain. The analgesic action of calcitonin may be mediated by restoration of serotonin receptors that control selective glutamate release from C-afferent fibers in ovariectomized rats and by normalization of sodium channel expression in damaged peripheral nerves. Serotonin receptors are reduced or eliminated by the relatively rapid reduction in estrogen during the postmenopausal period, and damaged nerves exhibit hyperexcitability due to abnormal expression of Na
Language	English
Publishing date	2017-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	2174252-2
ISSN	1744-8069 ; 1744-8069
ISSN (online)	1744-8069
ISSN	1744-8069
DOI	10.1177/1744806917720316
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Article: Characterization on Responsiveness of Excitatory Synaptic Transmissions to α1-Adrenoceptor Blockers in Substantia Gelatinosa Neurons Isolated From Lumbo-Sacral Level in Rat Spinal Cords.

Uta, Daisuke / Hattori, Tsuyoshi / Yoshimura, Megumu

International neurourology journal

2019 Volume 23, Issue 1, Page(s) 13–21

Abstract: Purpose: The aim of this study was to characterize the responsiveness of miniature excitatory postsynaptic currents (mEPSCs) to α1-adrenoceptor blockers in substantia gelatinosa (SG) neurons from the spinal cord to develop an explanation for the ... ...

Abstract	Purpose: The aim of this study was to characterize the responsiveness of miniature excitatory postsynaptic currents (mEPSCs) to α1-adrenoceptor blockers in substantia gelatinosa (SG) neurons from the spinal cord to develop an explanation for the efficacy of α1-adrenoceptor blockers in micturition dysfunction. Methods: Male adult Sprague-Dawley rats were used. Blind whole-cell patch-clamp recordings were performed using SG neurons in spinal cord slices. Naftopidil (100μM), tamsulosin (100μM), or silodosin (30μM), α1-adrenoceptor blockers, was perfused. The frequency of mEPSCs was recorded in an SG neuron to which the 3 blockers were applied sequentially with wash-out periods. Individual frequencies in a pair before naftopidil and tamsulosin perfusion were plotted as baseline, and the correlation between them was confirmed by Spearman correlation coefficient; linear regression was then performed. The same procedure was performed before naftopidil and silodosin perfusion. Frequencies of pairs after naftopidil and tamsulosin perfusion and after naftopidil and silodosin perfusion were similarly analyzed. The ratios of the frequencies after treatment to before were then calculated. Results: After the treatments, Spearman ρ and the slope were decreased to 0.682 from 0.899 at baseline and 0.469 from 1.004 at baseline, respectively, in the tamsulosin group relative to the naftopidil group. In the silodosin group, Spearman ρ and the slope were also decreased to 0.659 from 0.889 at baseline and 0.305 from 0.989 at baseline, respectively, relative to the naftopidil group. Naftopidil significantly increased the ratio of the frequency of mEPSCs compared to tamsulosin and silodosin (P=0.015 and P=0.004, respectively). Conclusion: There was a difference in responsiveness in the frequency of mEPSCs to α1-adrenoceptor blockers, with the response to naftopidil being the greatest among the α1-adrenoceptor blockers. These data are helpful to understand the action mechanisms of α1-adrenoceptor blockers for male lower urinary tract symptoms in clinical usage.
Language	English
Publishing date	2019-03-31
Publishing country	Korea (South)
Document type	Journal Article
ZDB-ID	2584447-7
ISSN	2093-6931 ; 2093-4777
ISSN (online)	2093-6931
ISSN	2093-4777
DOI	10.5213/inj.1938056.028
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Article ; Online: Chronic pain models amplify transient receptor potential vanilloid 1 (TRPV1) receptor responses in adult rat spinal dorsal horn.

Uta, Daisuke / Yoshimura, Megumu / Koga, Kohei

Neuropharmacology

2019 Volume 160, Page(s) 107753

Abstract: Persistent pain is associated with negative affect originating from hypersensitivity and/or allodynia. The spinal cord is a key area for nociception as well as chronic pain processing. Specifically, the dorsal horn neurons in lamina II (substantia ... ...

Abstract	Persistent pain is associated with negative affect originating from hypersensitivity and/or allodynia. The spinal cord is a key area for nociception as well as chronic pain processing. Specifically, the dorsal horn neurons in lamina II (substantia gelatinosa: SG) receive nociceptive inputs from primary afferents such as C fibers and/or Aδ fibers. Transient receptor potential vanilloid 1 (TRPV1) is a major receptor to sense heat as well as nociception. TRPV1 are expressed in the periphery and the central axon terminals of C fibers and/or Aδ fibers in the spinal cord. Activating TRPV1 enhances the release of glutamate in the spinal cord from naïve rodents. Here, we studied whether or not chronic pain could alter the response of TRPV1 channels to exogenous, capsaicin through study of synaptic transmission and neural activity in rat SG neurons. Using in vitro whole-cell patch-clamp recording, we found that bath application of capsaicin facilitated both the frequency and amplitude of miniature and spontaneous excitatory postsynaptic currents beyond a nerve injury and a complete Freund's adjuvant injection observed in the naïve group. Strikingly, capsaicin produced larger amplitudes of inward currents in pain models than compared to the naïve group. By contrast, the proportions of neurons that show capsaicin-induced inward currents were similar among naïve and pain groups. Importantly, the capsaicin-induced inward currents were conducted by TRPV1 and required calcium influx that was independent of voltage-gated calcium channels. Our study provides fundamental evidence that chronic inflammation and neuropathic pain models amplify the release of glutamate through the activation of TRPV1 in central axon terminals, and that facilitation of TRPV1 function in rat spinal SG neurons may contribute to enhanced capsaicin-induced inward currents.
MeSH term(s)	Animals ; Capsaicin/pharmacology ; Chronic Pain/drug therapy ; Chronic Pain/metabolism ; Disease Models, Animal ; Excitatory Postsynaptic Potentials/drug effects ; Freund's Adjuvant/pharmacology ; Inflammation/metabolism ; Male ; Patch-Clamp Techniques ; Posterior Horn Cells/metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Dorsal Horn/drug effects ; Spinal Cord Dorsal Horn/metabolism ; Substantia Gelatinosa/drug effects ; Substantia Gelatinosa/metabolism ; Synaptic Transmission/drug effects ; TRPV Cation Channels/drug effects ; TRPV Cation Channels/metabolism
Chemical Substances	TRPV Cation Channels ; Trpv1 protein, rat ; Freund's Adjuvant (9007-81-2) ; Capsaicin (S07O44R1ZM)
Language	English
Publishing date	2019-09-04
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	218272-5
ISSN	1873-7064 ; 0028-3908
ISSN (online)	1873-7064
ISSN	0028-3908
DOI	10.1016/j.neuropharm.2019.107753
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Article: Noradrenaline modulates mechanically evoked responses in the rat spinal dorsal horn: an in vivo patch-clamp study.

Sonohata, Motoki / Doi, Atsushi / Yasaka, Toshiharu / Uta, Daisuke / Mawatari, Masaaki / Yoshimura, Megumu

Journal of pain research

2019 Volume 12, Page(s) 1269–1278

Abstract: Purpose: ...

Abstract	Purpose:
Language	English
Publishing date	2019-04-17
Publishing country	New Zealand
Document type	Journal Article
ZDB-ID	2495284-9
ISSN	1178-7090
ISSN	1178-7090
DOI	10.2147/JPR.S181210
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