Elie D. Al-Chaer

Is there a pathway in the posterior funiculus that signals visceral pain

&NA; The present report provides evidence that axons in the medial part of the posterior column at T10 convey ascending nociceptive signals from pelvic visceral organs. This evidence was obtained from human surgical case studies and histological verification of the lesion in one of these cases, along with neuroanatomical and neurophysiological findings in animal experiments. A restricted lesion in this area can virtually eliminate pelvic pain due to cancer. The results remain excellent even in cases in which somatic structures of the pelvic body wall are involved. Following this procedure, neurological testing reveals no additional neurological deficit. There is no analgesia to pinprick stimuli applied to the body surface, despite the relief of the visceral pain. Since it is reasonable to attribute the favorable results of limited midline myelotomies to the interruption of axons of visceral nociceptive projection neurons in the posterior column, we have performed experiments in rats to test this hypothesis. The results in rats indicate that the dorsal column does indeed include a nociceptive component that signals pelvic visceral pain. The pathway includes neurons of the postsynaptic dorsal column pathway at the L6‐S1 segmental level, axons of these neurons in the fasciculus gracilis, and neurons of the nucleus gracilis and the ventral posterolateral nucleus of the thalamus.

Long-term sensitization of primary afferents in adult rats exposed to neonatal colon pain

We have previously shown that colon irritation (CI) in neonates results in chronic visceral hypersensitivity in adult rats, associated with central neuronal sensitization in the absence of identifiable peripheral pathology. The purpose of this study is to assess the relative contribution of peripheral mechanisms to chronic visceral hypersensitivity by examining the changes in responses of primary afferents at thoracolumbar (TL) and lumbosacral (LS) spinal segments to graded colorectal distension (CRD). Afferent discharges were recorded at the cut distal ends of spinal dorsal roots (DRs) in adult control and CI rats. We found that: (1). the average threshold of activation of LS afferents decreased significantly in CI rats; (2). the responses of TL and LS afferents to CRD in CI rats were significantly greater than those in control; (3). the spontaneous activity of LS afferents in CI rats was significantly stronger than that in control; (4). in CI rats, the average responses to graded CRD of LS DRs were significantly higher than those of TL DRs; and (5). the number of both LS DRs and TL DRs activated by CRD in CI rats was significantly larger than control. In summary, the results show that chronic visceral hypersensitivity is associated with peripheral sensitization, as well as central sensitization. TL visceral afferents projecting seem to be more involved in the processing of sensitized nociceptive input from the colon than acute nociceptive input. However, LS afferents seem to be equally important in both sensitized and acute pain states.

Sensitization of postsynaptic dorsal column neuronal responses by colon inflammation.

THE role of a newly identified component of the post-synaptic dorsal column (PSDC) system in viscerosensory processing has been recently described. The purpose of this study was to examine the effect of colon inflammation on the responses of single PSDC cells, located in the vicinity of the central canal at L6-S1 spinal segments, to graded colorectal distension (CRD) and to cutaneous stimulation. Experiments were conducted on seven male Sprague-Dawley rats anesthetized with pentobarbital. Recordings were made from seven PSDC cells located around the central canal at L6-S1 in response to CRD and cutaneous stimulation before and after colon inflammation. Inflammation of the colon with mustard oil (MO) induced an increase in the background activity of these cells. Colon inflammation also potentiated the responses of the PSDC cells to graded CRD but not to cutaneous stimulation. This is consistent with previously observed effects of colon inflammation on the responses of viscerosensitive cells in the ventral posterolateral (VPL) nucleus of the thalamus and in the nucleus gracilis (NG). These observations support a role of the PSDC system in viscerosensory processing and primary visceral hyperalgesia.

Potentiation of thalamic responses to colorectal distension by visceral inflammation.

Responses of single viscerosensitive units in the ventral posterolateral (VPL) nucleus of the thalamus to cutaneous stimuli and graded colorectal distension (CRD) were recorded before and after colon inflammation by mustard oil (MO). The spontaneous firing rate increased around 25 min after the injection of MO. Responses to CRD obtained after this time increased significantly. In contrast, responses to cutaneous stimuli were attenuated and no substantial variations in the size of the cutaneous receptive fields were observed. A lesion of the dorsal column (DC) at T10 dramatically decreased the background activity and almost totally abolished the responses to CRD and to innocuous cutaneous stimuli but did not significantly affect the responses to noxious cutaneous pinch. These results indicate that colon inflammation potentiates thalamic responses to visceral stimuli while decreasing those to cutaneous stimuli.

Microglia: a newly discovered role in visceral hypersensitivity?

Given the growing body of evidence for a role of glia in pain modulation, it is plausible that the exaggerated visceral pain in chronic conditions might be regulated by glial activation. In this study, we have investigated a possible role for microglia in rats with chronic visceral hypersensitivity and previously documented altered neuronal function. Experiments were performed on adult male Sprague-Dawley rats pre-treated with neonatal colon irritation (CI) and on control rats. Effects of fractalkine (FKN, a chemokine involved in neuron-to-microglia signaling) and of minocycline (an inhibitor of microglia) on visceral sensitivity were examined. Visceral sensitivity was assessed by recording the electromyographic (EMG) responses to graded colorectal distension (CRD) in mildly sedated rats. Responses to CRD were recorded before and after injection of FKN, minocycline or vehicle. Somatic thermal hyperalgesia was measured by latency of paw withdrawal to radiant heat. The pattern and intensity of microglial distribution at L6-S2 in the spinal cord was also compared in rats with CI and controls by fluorescence microscopy using OX-42. Results show that: (1) FKN significantly facilitated EMG responses to noxious CRD by >52% in control rats. FKN also induced thermal hyperalgesia in control rats, consistent with previous reports; (2) minocycline significantly inhibited EMG responses to noxious CRD by >70% in rats with CI compared to controls 60 min after injection. The anti-nociceptive effect of minocycline lasted for 180 min in rats with CI, reaching peak values 60 min after injection. Our results show that FKN enhances visceral and somatic nociception, whereas minocycline inhibits visceral hypersensitivity in chronically sensitized rats, which indicates a role for microglia in visceral hypersensitivity.

Thalamic modulation of visceral nociceptive processing in adult rats with neonatal colon irritation

Visceral pain originates from visceral organs in response to a noxious stimulus which, if prolonged, may lead to chronic changes in the neural network mediating visceral nociception. For instance, colon inflammation enhances the responses of neurons in the thalamus to colorectal distension (CRD), whereas lesion in the dorsal column (DC) reverses this neuronal sensitization, suggesting that the thalamus and the DC play major roles in chronic visceral pain. In this study, we used adult rats sensitized with neonatal painful colon irritation to reveal the contribution of the thalamus and the DC to neuronal hyperexcitability in a model of chronic visceral pain. We recorded the responses of lumbosacral neurons to CRD in control rats and in rats with colon irritation following stimulation or inactivation of the thalamus, and after DC lesion. Our results show that, first, neuronal responses to CRD decreased following thalamic stimulation in control rats, whereas, in rats with colon irritation, responses either decreased or increased; second, DC lesion attenuated or enhanced these effects in the positively or in the negatively modulated group of neurons, respectively; third, lidocaine injection in the thalamus reduced the responses to CRD in some of the neurons recorded in rats with colon irritation, but had no effect on those in control rats. Therefore, it is reasonable to speculate that plasticity in rats with colon irritation that may underlie chronic pain is sustained by feedback loops ascending in the DC and engaging the thalamus.

COMPARISON OF METABOLIC AND NEUROPATHY PROFILES OF RATS WITH STREPTOZOTOCIN-INDUCED OVERT AND MODERATE INSULINOPENIA

To assess the relative roles of insulinopenia, hyperglycemia and dyslipidemia in pathogenesis of diabetic neuropathy, we compared plasma insulin, glucose and lipid metabolism and peripheral nerve function in rats with streptozotocin (STZ)-induced overt and moderate insulinopenia (hyperglycemic, STZ-HG; random glucose>11 mM and normoglycemic, STZ-NG rats). While being slightly insulinopenic, STZ-NG rats are metabolically not different from control, naive animals, by having normal glucose tolerance and normal levels of plasma glucose, glycated HbA1c, cholesterol and triglycerides. Two weeks following injection of STZ, STZ-HG but not STZ-NG rats had suppressed motor nerve conduction velocity, F-wave prevalence, withdrawal responses to heat and von Frey filament stimuli. In apparent correlation with plasma insulin level, both STZ-HG and -NG rats manifested exaggerated responses in paw pressure and colorectal distension tests. These data suggest that insulinopenia may play a leading role in the diabetic impairment of deep muscle and visceral afferent pathways while hyperglycemia/dyslipidemia may represent a key requirement for the onset and progression of electrophysiological nerve impairment and loss of superficial heat and tactile perception. STZ-NG rats offer a convenient model for the investigation of the short-term effects of insulinopenia on peripheral nerve function.

Altered behavior and digestive outcomes in adult male rats primed with minimal colon pain as neonates

BackgroundNeonatal colon irritation (CI; pain or inflammation) given for 2 weeks prior to postnatal day 22 (PND22), causes long-lasting functional disorders in rats that can be seen 6 months after the initial insult. This study looked at the effect of varying the frequency and duration of neonatal CI on the rate of growth, digestive outcomes, exploratory activity, and colon and skin sensitivity in adult rats.MethodsMale Sprague-Dawley rats were given CI using repeated colorectal distension (CRD) at different time intervals and for varying durations starting at PND 8, 10 or 14. Control rats were handled by the investigator without any intracolonic insertion. Further experiments were done on adult rats. Digestive outcomes (food and water consumption, fecal and urinary outputs) were measured using metabolic cages. Exploratory behavior was measured using digital video tracking in an open field. Cutaneous sensitivity was assessed by measuring the responses to mechanical and heat stimuli applied to the shaved abdomen or hind paws. Visceral sensitivity was measured by recording electromyographic responses, under light isoflurane anesthesia, from the external oblique muscles in response to CRD.ResultsNo significant weight differences were observed between CI and control rats. Exploratory behavior was reduced in rats with neonatal CI compared to control. Digestive outputs and somatic and visceral sensitivity changed between different treatment groups with earlier and more frequent insults yielding a higher deviation from normal.ConclusionThe diversity of behavioral and digestive symptoms in these rats parallels the diversity of symptoms in patients with functional gastrointestinal disorders and is consistent with global plastic changes affecting more than one system in the organism.

Noxious visceral inputs enhance cutaneous tactile response in rat thalamus

The current study investigates whether visceral nociceptive inputs affect tactile processing in the thalamic ventroposterior lateral nucleus in anesthetized rats by means of extracellular single unit recordings. Twenty out of the 44 neurons had their response to tactile stimulation increased by preceding nociceptive colorectal distension (CRD), and this influence appear more potent than the opposite effect, tactile on CRD response. There was a dynamic change of tactile response along with CRD response and the background activities as a function of repeated colorectal stimulation. The abnormal neuronal discharge in response to tactile stimulation in the condition of visceral nociception could contribute to the development of referred pain and allodynia.

Importance of neural mechanisms in colonic mucosal and muscular dysfunction in adult rats following neonatal colonic irritation

Previous studies have shown that early life trauma induced by maternal separation or colonic irritation leads to hypersensitivity to colorectal distension in adulthood. We tested the hypothesis that repetitive colorectal distension in neonates leads to abnormalities in colonic permeability and smooth muscle function in the adult rat. In neonatal rats, repetitive colorectal distension was performed on days 8, 10, and 12. As adults, stool consistency was graded from 0 (formed stool) to 3 (liquid stool). Colonic tissue was isolated for histology and myeloperoxidase levels. The colonic mucosa was placed in modified Ussing chambers for measurements of permeability and short‐circuit current responses to forskolin, electrical field stimulation, and carbachol. Segments of colonic musculature were placed in organ baths and contractile response to potassium chloride, electrical field stimulation, and carbachol were determined. In adult rats that experienced neonatal colonic irritation, no significant changes in colonic histology or myeloperoxidase activity were observed; however, stool consistency scores were increased. Mucosal permeability, measured as an increase in basal conductance, was significantly increased but no changes in short‐circuit current responses were observed. In adulthood, rats that underwent colorectal distension as neonates exhibited an elevated smooth muscle contractile response to potassium chloride, but no changes in response to electrical field stimulation or carbachol. In summary, neonatal colonic irritation, shown previously to produce colonic hypersensitivity, leads to significant alterations in colonic mucosal and smooth muscle function characterized by loose stools, increased mucosal permeability, and increased smooth muscle contractility in the absence of colon inflammation in adulthood.