Dual purinergic synaptic transmission in the human enteric nervous system

J. E. Wunderlich, B. J. Needleman, Z. Chen, J. G. Yu, Y. Wang, I. Grants, D. J. Mikami, W. Scott Melvin, H. J. Cooke, F. L. Christofi

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Based on findings in rodents, we sought to test the hypothesis that purinergic modulation of synaptic transmission occurs in the human intestine. Time series analysis of intraneuronal free Ca2+ levels in submucosal plexus (SMP) from Roux-en-Y specimens was done using Zeiss LSM laser-scanning confocal fluo-4 AM Ca2+ imaging. A 3-s fiber tract stimulation (FTS) was used to elicit a synaptic Ca2+ response. Short-circuit current (Isc = chloride secretion) was recorded in mucosa-SMP in flux chambers. A distension reflex or electrical field stimulation was used to study Isc responses. Ca2+ imaging was done in 1,222 neurons responding to high-K+ depolarization from 61 surgical cases. FTS evoked synaptic Ca2+ responses in 62% of recorded neurons. FTS caused frequency-dependent Ca2+ responses (0.1-100 Hz). FTS Ca2+ responses were inhibited by Ω-conotoxin (70%), hexamethonium (50%), TTX, high Mg2+/low Ca2+ (≤100%), or capsaicin (25%). A P2Y1 receptor (P2Y1R) antagonist, MRS-2179 or PLC inhibitor U-73122, blocked FTS responses (75-90%). P2Y1R- immunoreactivity occurred in 39% of vasoactive intestinal peptide-positive neurons. The selective adenosine A3 receptor (AdoA3R) agonist 2-chloro-N6-(3-iodobenzyl)adenosine-5′-N- methylcarboxamide (2-Cl-IBMECA) caused concentration- and frequency-dependent inhibition of FTS Ca2+ responses (IC50 = 8.5 × 10-8 M). The AdoA3R antagonist MRS-1220 augmented such Ca2+ responses; 2-Cl-IBMECA competed with MRS-1220. Knockdown of AdoA1R with 8-cyclopentyl-3-N-(3-{[3-(4-fluorosulphonyl)benzoyl]-oxy} -propyl)-1-N-propyl-xanthine did not prevent 2-Cl-IBMECA effects. MRS-1220 caused 31% augmentation of TTX-sensitive distension Isc responses. The SMP from Roux-en-Y patients is a suitable model to study synaptic transmission in human enteric nervous system (huENS). The P2Y 1/Gαq/PLC/inositol 1,3,5-trisphosphate/Ca2+ signaling pathway, N-type Ca2+ channels, nicotinic receptors, and extrinsic nerves contribute to neurotransmission in huENS. Inhibitory AdoA 3R inhibit nucleotide or cholinergic transmission in the huENS.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume294
Issue number2
DOIs
StatePublished - Feb 2008
Externally publishedYes

Fingerprint

Enteric Nervous System
Submucous Plexus
Synaptic Transmission
Purinergic P2Y1 Receptors
Neurons
Adenosine A3 Receptor Antagonists
Adenosine A3 Receptor Agonists
Conotoxins
Hexamethonium
Xanthine
Capsaicin
Vasoactive Intestinal Peptide
Nicotinic Receptors
Inositol
Cholinergic Agents
Electric Stimulation
Inhibitory Concentration 50
Intestines
Reflex
Chlorides

Keywords

  • Adenosine A receptors
  • Calcium signaling
  • P2Y receptors
  • Purinergic transmission
  • Submucous nerve plexus

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology

Cite this

Dual purinergic synaptic transmission in the human enteric nervous system. / Wunderlich, J. E.; Needleman, B. J.; Chen, Z.; Yu, J. G.; Wang, Y.; Grants, I.; Mikami, D. J.; Melvin, W. Scott; Cooke, H. J.; Christofi, F. L.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 294, No. 2, 02.2008.

Research output: Contribution to journalArticle

Wunderlich, JE, Needleman, BJ, Chen, Z, Yu, JG, Wang, Y, Grants, I, Mikami, DJ, Melvin, WS, Cooke, HJ & Christofi, FL 2008, 'Dual purinergic synaptic transmission in the human enteric nervous system', American Journal of Physiology - Gastrointestinal and Liver Physiology, vol. 294, no. 2. https://doi.org/10.1152/ajpgi.00500.2007
Wunderlich, J. E. ; Needleman, B. J. ; Chen, Z. ; Yu, J. G. ; Wang, Y. ; Grants, I. ; Mikami, D. J. ; Melvin, W. Scott ; Cooke, H. J. ; Christofi, F. L. / Dual purinergic synaptic transmission in the human enteric nervous system. In: American Journal of Physiology - Gastrointestinal and Liver Physiology. 2008 ; Vol. 294, No. 2.
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AU - Needleman, B. J.

AU - Chen, Z.

AU - Yu, J. G.

AU - Wang, Y.

AU - Grants, I.

AU - Mikami, D. J.

AU - Melvin, W. Scott

AU - Cooke, H. J.

AU - Christofi, F. L.

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N2 - Based on findings in rodents, we sought to test the hypothesis that purinergic modulation of synaptic transmission occurs in the human intestine. Time series analysis of intraneuronal free Ca2+ levels in submucosal plexus (SMP) from Roux-en-Y specimens was done using Zeiss LSM laser-scanning confocal fluo-4 AM Ca2+ imaging. A 3-s fiber tract stimulation (FTS) was used to elicit a synaptic Ca2+ response. Short-circuit current (Isc = chloride secretion) was recorded in mucosa-SMP in flux chambers. A distension reflex or electrical field stimulation was used to study Isc responses. Ca2+ imaging was done in 1,222 neurons responding to high-K+ depolarization from 61 surgical cases. FTS evoked synaptic Ca2+ responses in 62% of recorded neurons. FTS caused frequency-dependent Ca2+ responses (0.1-100 Hz). FTS Ca2+ responses were inhibited by Ω-conotoxin (70%), hexamethonium (50%), TTX, high Mg2+/low Ca2+ (≤100%), or capsaicin (25%). A P2Y1 receptor (P2Y1R) antagonist, MRS-2179 or PLC inhibitor U-73122, blocked FTS responses (75-90%). P2Y1R- immunoreactivity occurred in 39% of vasoactive intestinal peptide-positive neurons. The selective adenosine A3 receptor (AdoA3R) agonist 2-chloro-N6-(3-iodobenzyl)adenosine-5′-N- methylcarboxamide (2-Cl-IBMECA) caused concentration- and frequency-dependent inhibition of FTS Ca2+ responses (IC50 = 8.5 × 10-8 M). The AdoA3R antagonist MRS-1220 augmented such Ca2+ responses; 2-Cl-IBMECA competed with MRS-1220. Knockdown of AdoA1R with 8-cyclopentyl-3-N-(3-{[3-(4-fluorosulphonyl)benzoyl]-oxy} -propyl)-1-N-propyl-xanthine did not prevent 2-Cl-IBMECA effects. MRS-1220 caused 31% augmentation of TTX-sensitive distension Isc responses. The SMP from Roux-en-Y patients is a suitable model to study synaptic transmission in human enteric nervous system (huENS). The P2Y 1/Gαq/PLC/inositol 1,3,5-trisphosphate/Ca2+ signaling pathway, N-type Ca2+ channels, nicotinic receptors, and extrinsic nerves contribute to neurotransmission in huENS. Inhibitory AdoA 3R inhibit nucleotide or cholinergic transmission in the huENS.

AB - Based on findings in rodents, we sought to test the hypothesis that purinergic modulation of synaptic transmission occurs in the human intestine. Time series analysis of intraneuronal free Ca2+ levels in submucosal plexus (SMP) from Roux-en-Y specimens was done using Zeiss LSM laser-scanning confocal fluo-4 AM Ca2+ imaging. A 3-s fiber tract stimulation (FTS) was used to elicit a synaptic Ca2+ response. Short-circuit current (Isc = chloride secretion) was recorded in mucosa-SMP in flux chambers. A distension reflex or electrical field stimulation was used to study Isc responses. Ca2+ imaging was done in 1,222 neurons responding to high-K+ depolarization from 61 surgical cases. FTS evoked synaptic Ca2+ responses in 62% of recorded neurons. FTS caused frequency-dependent Ca2+ responses (0.1-100 Hz). FTS Ca2+ responses were inhibited by Ω-conotoxin (70%), hexamethonium (50%), TTX, high Mg2+/low Ca2+ (≤100%), or capsaicin (25%). A P2Y1 receptor (P2Y1R) antagonist, MRS-2179 or PLC inhibitor U-73122, blocked FTS responses (75-90%). P2Y1R- immunoreactivity occurred in 39% of vasoactive intestinal peptide-positive neurons. The selective adenosine A3 receptor (AdoA3R) agonist 2-chloro-N6-(3-iodobenzyl)adenosine-5′-N- methylcarboxamide (2-Cl-IBMECA) caused concentration- and frequency-dependent inhibition of FTS Ca2+ responses (IC50 = 8.5 × 10-8 M). The AdoA3R antagonist MRS-1220 augmented such Ca2+ responses; 2-Cl-IBMECA competed with MRS-1220. Knockdown of AdoA1R with 8-cyclopentyl-3-N-(3-{[3-(4-fluorosulphonyl)benzoyl]-oxy} -propyl)-1-N-propyl-xanthine did not prevent 2-Cl-IBMECA effects. MRS-1220 caused 31% augmentation of TTX-sensitive distension Isc responses. The SMP from Roux-en-Y patients is a suitable model to study synaptic transmission in human enteric nervous system (huENS). The P2Y 1/Gαq/PLC/inositol 1,3,5-trisphosphate/Ca2+ signaling pathway, N-type Ca2+ channels, nicotinic receptors, and extrinsic nerves contribute to neurotransmission in huENS. Inhibitory AdoA 3R inhibit nucleotide or cholinergic transmission in the huENS.

KW - Adenosine A receptors

KW - Calcium signaling

KW - P2Y receptors

KW - Purinergic transmission

KW - Submucous nerve plexus

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