Matthew James Broadhead

Publications (3) View all

• Article: Ca(2+) transients in submucous neurons during the colonic migrating motor complex in the isolated murine large intestine.

  T Okamoto, P O Bayguinov, M J Broadhead, T K Smith
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  ABSTRACT: The colonic migrating motor complex (CMMC) is a spontaneous, rhythmic, and neurally mediated motor pattern generated by myenteric neurons, which can propel fecal pellets in mice. Our aim was to determine whether submucous neurons were also activated during the CMMC. :The isolated murine colon was opened and sections of mucosa were removed to expose the submucous ganglia, which were then loaded with Fluo-4. Colonic migrating motor complexes, which occurred spontaneously or by mechanically stimulating the mucosa, were identified by displacement of the tissue (duration = 23.3 s). Between CMMCs, spontaneous Ca(2+) transients (frequency = 0.9 Hz) were observed in 55% (n = 8) of submucous neurons. During the CMMC, 98% (seven ganglia, n = 7) of submucous neurons within the same ganglion exhibited rapid Ca(2+) transients (1.6 Hz) superimposed on a sustained rise in Ca(2+) (duration ∼23 s) that occurred 1.7 s following the mucosal stimulus; whereas other neurons exhibited a similar, but delayed response that occurred either at 7 or 13 s following the stimulus. The activity in submucous neurons was correlated with activity in adjacent nerve varicosities. Ondansetron (1 mm; 5-HT(3) antagonist) significantly reduced the frequency and duration of the Ca(2+) transient responses. Activity in the submucous neurons appears to be secondary to that in the myenteric plexus and appears to be generated largely by activity in myenteric descending (serotonergic) interneurons. During the CMMC, there is likely to be an increase in secretion to lubricate and facilitate fecal pellet propulsion.
  Neurogastroenterology and Motility 05/2012; 24(8):769-78, e354. · 3.41 Impact Factor
• Article: Activity in varicosities within the myenteric plexus between and during the colonic migrating motor complex in the isolated murine large intestine.

  P O Bayguinov, M J Broadhead, T Okamoto, G W Hennig, T K Smith
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  ABSTRACT:   Neuronal communication within the myenteric plexus occurs when action potentials along nerve fibers produce Ca(2+) transients in varicosities leading to exocytosis of vesicles and neurotransmitters release. We used Ca(2+) transients in varicosities to monitor action potential activity in myenteric nerve pathways both between and during the colonic migrating motor complex (CMMC) in the isolated murine colon.   Strips of longitudinal muscle were removed to reveal the myenteric ganglia, which were then loaded with Fluo-4.   Many varicosities, including synaptotagmin 1 labeled varicosities, exhibited ongoing Ca(2+) transients (duration of unitary Ca(2+) transient 3.9 s). Between CMMCs, varicosities fired at a frequency of 0.6 Hz, which correlated with spontaneous inhibitory junction potentials in the circular muscle, suggesting they were mainly in inhibitory nerve pathways. During a CMMC other previously quiescent varicosities fired at 1.3 Hz (max. 2.0 Hz) for the duration (24 s) of the CMMC, suggesting they were on excitatory nerve pathways. Activity in varicosities was correlated with Ca(2+) transient responses in a number of neurons. Some varicosities appeared to release an inhibitory neurotransmitter that reduced activity in nNOS-positive neurons. Varicosities along the same nerve fiber exhibited identical patterns of activity that allowed nerve fibers to be traced throughout the myenteric plexus and internodal strands. Activity in varicosities was reduced by hexamethonium (100 μmol L(-1) ), and blocked by ω-conotoxin GVIA (200 nM) and tetrodotoxin (1 μmol L(-1) ; TTX).   Ca(2+) imaging of varicosities allows for a determination of activity in neural pathways within the enteric nervous system.
  Neurogastroenterology and Motility 02/2012; 24(4):e185-201. · 3.41 Impact Factor
• Article: Ca2+ transients in myenteric glial cells during the colonic migrating motor complex in the isolated murine large intestine.

  Matthew J Broadhead, Peter O Bayguinov, Takanobu Okamoto, Dante J Heredia, Terence K Smith
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  ABSTRACT: Enteric glia cells (EGCs) form a dense network around myenteric neurons in a ganglia and are likely to have not only a supportive role but may also regulate or be regulated by neural activity. Our aims were to determine if EGCs are activated during the colonic migrating motor complex (CMMC) in the isolated murine colon. Strips of longitudinal muscle were removed and Ca(2+) imaging (Fluo-4) used to study activity in EGCs within myenteric ganglia during CMMCs, followed by post hoc S100 staining to reveal EGCs. The cell bodies of EGCs and their processes formed caps and halos, respectively, around some neighbouring myenteric neurons. Some EGCs (36%), which were largely quiescent between CMMCs, exhibited prolonged tetrodotoxin (TTX; 1 μm)-sensitive Ca(2+) transients that peaked ∼39 s following a mucosal stimulus that generated the CMMC, and often outlasted the CMMC (duration ∼23 s). Ca(2+) transients in EGCs often varied in duration within a ganglion; however, the duration of these transients was closely matched by activity in closely apposed nerve varicosities, suggesting EGCs were not only innervated but the effective innervation was localized. Furthermore, all EGCs, even those that were quiescent, responded with robust Ca(2+) transients to KCl, caffeine, nicotine, substance P and GR 64349 (an NK2 agonist), suggesting they were adequately loaded with indicator and that some EGCs may be inhibited by substances released by neighbouring neurons. Intracellular Ca(2+) waves were visualised propagating between closely apposed glia and from glial cell processes to the soma (velocity 12 μm s(-1)) where they produced an accumulative rise in Ca(2+), suggesting that the soma acts as an integrator of Ca(2+) activity. In conclusion, Ca(2+) transients in EGCs occur secondary to nerve activity; their activation is driven by intrinsic excitatory nerve pathways that generate the CMMC.
  The Journal of Physiology 11/2011; 590(Pt 2):335-50. · 4.72 Impact Factor