Biomedical subjects

H Ennes

Publications and source records attributed to H Ennes.

11 recordsLinked to original sources

PubMed2001

Agonists of proteinase-activated receptor 1 induce plasma extravasation by a neurogenic mechanism.

Thrombin, generated in the circulation during injury, cleaves proteinase-activated receptor 1 (PAR1) to stimulate plasma extravasation and granulocyte infiltration. However, the mechanism of thrombin-induced inflammation in intact tissues is unknown. We hypothesized that thrombin cleaves PAR1 on sensory nerves to release substance P (SP), which interacts with the neurokinin 1 receptor (NK1R) on endothelial cells to cause plasma extravasation. PAR1 was detected in small diameter neurons known to contain SP in rat dorsal root ganglia by immunohistochemistry and in situ hybridization. Thrombin and the PAR1 agonist TFLLR-NH(2) (TF-NH(2)) increased [Ca(2+)](i) >50% of cultured neurons (EC(50)s 24 mu ml(-1) and 1.9 microM, respectively), assessed using Fura-2 AM. The PAR1 agonist completely desensitized responses to thrombin, indicating that thrombin stimulates neurons through PAR1. Injection of TF-NH(2) into the rat paw stimulated a marked and sustained oedema. An NK1R antagonist and ablation of sensory nerves with capsaicin inhibited oedema by 44% at 1 h and completely by 5 h. In wild-type but not PAR1(-/-) mice, TF-NH(2) stimulated Evans blue extravasation in the bladder, oesophagus, stomach, intestine and pancreas by 2 - 8 fold. Extravasation in the bladder, oesophagus and stomach was abolished by an NK1R antagonist. Thus, thrombin cleaves PAR1 on primary spinal afferent neurons to release SP, which activates the NK1R on endothelial cells to stimulate gap formation, extravasation of plasma proteins, and oedema. In intact tissues, neurogenic mechanisms are predominantly responsible for PAR1-induced oedema.

Animals↗

PubMed1999

Involvement of stretch-sensitive calcium flux in mechanical transduction in visceral afferents.

The spinal and vagal visceral innervation to the gastrointestinal tract contains mechanosensitive afferents that are activated by contraction, distension of smooth muscle or movement in the receptive field. The mechanism by which free nerve endings detect changes in smooth muscle tension is not clear. The present study investigated the effects of mechanical stimulation on dorsal root ganglion neurons in vitro. Neurons were cultured using standard techniques and used in experiments after 24-72 h. Intracellular calcium [Ca2+]i was visualized using a video microscopic technique (Attoflour) in Fura-2 loaded neurons. DRG neurons innervating the stomach or colon were identified by the presence of a retrograde tracer, dextran-conjugated Texas Red, injected into the visceral wall 14-28 days previously. Increases in [Ca2+]i were measured in response to transient (0.5 s) mechanical stimulation of the cell soma using a flame polished probe. Approximately 25% of the whole population of DRG neurons (n = 199) were mechanosensitive, showing a transient rise in [Ca2+]i. In labeled afferents (n = 12), approximately 40% of neurons were mechanosensitive. The increase in [Ca2+]i in response to mechanical stimulation was reduced (100 microM) or abolished (250 microM) by superfusion with gadolinium or by removal of extracellular calcium. Cell somata of visceral spinal afferents show a stretch-sensitive calcium flux that may be involved in sensory transduction of mechanical stimuli that lead to autonomic and sensory reflexes.

Animals↗

PubMed1997

Regional cerebral activity in normal and pathological perception of visceral pain.

BACKGROUND & AIMS: To characterize the cerebral processing of noxious visceral events, changes in regional cerebral blood flow associated with perception of intestinal pain were examined. METHODS: The effects of rectal pressure stimuli on regional cerebral blood flow were assessed with 15O-water positron emission tomography (PET) in 12 subjects, half with irritable bowel syndrome (IBS). PET scans were obtained at baseline and during both actual and simulated delivery of anticipated stimuli. Changes in regional cerebral blood flow were interpreted using statistical parametric mapping and region of interest methods of analysis. RESULTS: In healthy subjects, perception of pain during actual or simulated delivery of painful stimuli was significantly associated (P < 0.01) with activity of the anterior cingulate cortex (ACC; Brodmann's areas 24 and 32), whereas no ACC response to perception of nonpainful stimuli was observed. In patients with IBS, the ACC failed to respond to the same stimuli, whereas significant activation (P < 0.01) of the left prefrontal cortex (maximal in Brodmann's area 10) was seen. CONCLUSIONS: The perception of acute rectal pain is associated with activation of the ACC in healthy subjects, and patients with IBS show an aberrant brain activation pattern both during noxious rectal distention and during the anticipation of rectal pain.

Adult↗

PubMed1992

Effect of cell culture on rabbit colonic smooth muscle bradykinin receptors.

The aim of this study was to assess the effect of cell culture on the bradykinin receptor of rabbit colon myocytes. In longitudinal muscle strips prepared from distal colon, bradykinin stimulated dose-dependent contraction that was 62% of the maximal response to bethanecol. At 4 degrees C, [3H]bradykinin binding to fresh muscle homogenates from the distal colon was time dependent, saturable, and linearly related to tissue concentration. Specific binding of 0.6 nmol/L [3H]bradykinin was 80% +/- 2% of total binding. In competitive binding studies, Hill coefficients approached unity, suggesting the presence of a single class of receptors. The order of potency was bradykinin greater than [D-Phe7]bradykinin much greater than des-Arg9, [Leu8]bradykinin, which is consistent with results of a B2 receptor subclass. Colon myocytes from the longitudinal muscle layer achieved confluence and were harvested for studies after 12-14 days in culture. Bradykinin receptors were of high affinity [disassociation constant (Kd) = 672 pmol/L] and numbered 10,217 +/- 2567/cell. To show that the receptors on cultured myocytes were functional, the effect of bradykinin was measured (a) on intracellular calcium concentration using Fura 2 and (b) on prostaglandin E2 concentration in the culture media using radioimmunoassay. In cells grown to confluence on cover slips and preloaded with Fura 2, bradykinin stimulated the threshold response at 1 nmol/L and maximal response (increased intracellular calcium concentration from 229 to 633 nmol/L) at 1 mumol/L. Bradykinin, 100 nmol/L, increased Prostaglandin E2 in the culture media threefold. In summary, colon myocytes express functioning bradykinin receptors, which, unlike muscarinic receptors, persist in culture. Bradykinin appears to be a suitable agonist for studies of receptor-mediated intracellular events in cultured colon myocytes.

Animals↗

PubMed1990

Substance P and CGRP mediate motor response of rabbit colon to capsaicin.

Primary afferent nerve terminals located in the mammalian gut wall may play a role in region-specific modulation of gastrointestinal motility. In the present study, we sought to characterize the effect of neuropeptides released from these afferents by capsaicin (CAP) on contractile activity of smooth muscle from the distal rabbit colon. CAP caused a release of acetylcholine and immunoreactivity for substance P (SP) and calcitonin gene-related peptide (CGRP) from the muscle coat. CAP caused a dose-dependent transient stimulation of longitudinal muscle contractions, followed by prolonged inhibition of spontaneous but not stimulated contractile activity. The initial stimulation was abolished by the SP antagonist spantide and by atropine. The inhibitory effect was reduced by repeated exposure of muscle to CGRP. The effect of CGRP on spontaneous contractions differed between longitudinal and circular muscle. In longitudinal muscle, a stimulation was preceded by a transient inhibition, whereas in circular muscle, only inhibition was seen. Both effects were resistant to tetrodotoxin. Repeated exposure of circular but not longitudinal muscle to CGRP resulted in a disappearance of the peptide's inhibitory effect. Exogenously applied CGRP was only a weak antagonist of contractions stimulated by SP and bethanechol. These findings suggest that in the rabbit colon at least the following two neuropeptides are released from CAP-sensitive nerve fibers: a neurokinin peptide from nerve terminals located within the myenteric plexus and CGRP from terminals probably located within the circular muscle layer.

Acetylcholine↗

PubMed1970