Pancreatic Afferents Research Articles

Nucleus tractus solitarius mediates hyperalgesia induced by chronic pancreatitis in rats.

BACKGROUNDCentral sensitization plays a pivotal role in the maintenance of chronic pain induced by chronic pancreatitis (CP). We hypothesized that the nucleus tractus solitarius (NTS), a primary central site that integrates pancreatic afferents apart from the thoracic spinal dorsal horn, plays a key role in the pathogenesis of visceral hypersensitivity in a rat model of CP.AIMTo investigate the role of the NTS in the visceral hypersensitivity induced by chronic pancreatitis.METHODSCP was induced by the intraductal injection of trinitrobenzene sulfonic acid (TNBS) in rats. Pancreatic hyperalgesia was assessed by referred somatic pain via von Frey filament assay. Neural activation of the NTS was indicated by immunohistochemical staining for Fos. Basic synaptic transmission within the NTS was assessed by electrophysiological recordings. Expression of vesicular glutamate transporters (VGluTs), N-methyl-D-aspartate receptor subtype 2B (NR2B), and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor subtype 1 (GluR1) was analyzed by immunoblotting. Membrane insertion of NR2B and GluR1 was evaluated by electron microscopy. The regulatory role of the NTS in visceral hypersensitivity was detected via pharmacological approach and chemogenetics in CP rats.RESULTSTNBS treatment significantly increased the number of Fos-expressing neurons within the caudal NTS. The excitatory synaptic transmission was substantially potentiated within the caudal NTS in CP rats (frequency: 5.87 ± 1.12 Hz in CP rats vs 2.55 ± 0.44 Hz in sham rats, P < 0.01; amplitude: 19.60 ± 1.39 pA in CP rats vs 14.71 ± 1.07 pA in sham rats; P < 0.01). CP rats showed upregulated expression of VGluT2, and increased phosphorylation and postsynaptic trafficking of NR2B and GluR1 within the caudal NTS. Blocking excitatory synaptic transmission via the AMPAR antagonist CNQX and the NMDAR antagonist AP-5 microinjection reversed visceral hypersensitivity in CP rats (abdominal withdraw threshold: 7.00 ± 1.02 g in CNQX group, 8.00 ± 0.81 g in AP-5 group and 1.10 ± 0.27 g in saline group, P < 0.001). Inhibiting the excitability of NTS neurons via chemogenetics also significantly attenuated pancreatic hyperalgesia (abdominal withdraw threshold: 13.67 ± 2.55 g in Gi group, 2.00 ± 1.37 g in Gq group, and 2.36 ± 0.67 g in mCherry group, P < 0.01).CONCLUSIONOur findings suggest that enhanced excitatory transmission within the caudal NTS contributes to pancreatic pain and emphasize the NTS as a pivotal hub for the processing of pancreatic afferents, which provide novel insights into the central sensitization of painful CP.

Neuroplastic Changes Occur Early in the Development of Pancreatic Ductal Adenocarcinoma

Perineural tumor invasion of intrapancreatic nerves, neurogenic inflammation, and tumor metastases along extrapancreatic nerves are key features of pancreatic malignancies. Animal studies show that chronic pancreatic inflammation produces hypertrophy and hypersensitivity of pancreatic afferents and that sensory fibers may themselves drive inflammation via neurogenic mechanisms. Although genetic mutations are required for cancer development, inflammation has been shown to be a precipitating event that can accelerate the transition of precancerous lesions to cancer. These observations led us to hypothesize that inflammation that accompanies early phases of pancreatic ductal adenocarcinoma (PDAC) would produce pathologic changes in pancreatic neurons and innervation. Using a lineage-labeled genetically engineered mouse model of PDAC, we found that pancreatic neurotrophic factor mRNA expression and sensory innervation increased dramatically when only pancreatic intraepithelial neoplasia were apparent. These changes correlated with pain-related decreases in exploratory behavior and increased expression of nociceptive genes in sensory ganglia. At later stages, cells of pancreatic origin could be found in the celiac and sensory ganglia along with metastases to the spinal cord. These results demonstrate that the nervous system participates in all stages of PDAC, including those that precede the appearance of cancer.

TRPV1 and TRPA1 Antagonists Prevent the Transition of Acute to Chronic Inflammation and Pain in Chronic Pancreatitis

Visceral afferents expressing transient receptor potential (TRP) channels TRPV1 and TRPA1 are thought to be required for neurogenic inflammation and development of inflammatory hyperalgesia. Using a mouse model of chronic pancreatitis (CP) produced by repeated episodes (twice weekly) of caerulein-induced AP (AP), we studied the involvement of these TRP channels in pancreatic inflammation and pain-related behaviors. Antagonists of the two TRP channels were administered at different times to block the neurogenic component of AP. Six bouts of AP (over 3 wks) increased pancreatic inflammation and pain-related behaviors, produced fibrosis and sprouting of pancreatic nerve fibers, and increased TRPV1 and TRPA1 gene transcripts and a nociceptive marker, pERK, in pancreas afferent somata. Treatment with TRP antagonists, when initiated before week 3, decreased pancreatic inflammation and pain-related behaviors and also blocked the development of histopathological changes in the pancreas and upregulation of TRPV1, TRPA1, and pERK in pancreatic afferents. Continued treatment with TRP antagonists blocked the development of CP and pain behaviors even when mice were challenged with seven more weeks of twice weekly caerulein. When started after week 3, however, treatment with TRP antagonists was ineffective in blocking the transition from AP to CP and the emergence of pain behaviors. These results suggest: (1) an important role for neurogenic inflammation in pancreatitis and pain-related behaviors, (2) that there is a transition from AP to CP, after which TRP channel antagonism is ineffective, and thus (3) that early intervention with TRP channel antagonists may attenuate the transition to and development of CP effectively.

Distribution and neurochemical identification of pancreatic afferents in the mouse

Dysfunction of primary afferents innervating the pancreas has been shown to contribute to the development of painful symptoms during acute and chronic pancreatitis. To investigate the distribution and neurochemical phenotype of pancreatic afferents, Alexa Fluor-conjugated cholera toxin B (CTB) was injected into the pancreatic head (CTB-488) and tail (CTB-555) of adult male mice to label neurons retrogradely in both the dorsal root ganglia (DRG) and nodose ganglia (NG). The NG and DRG (T5-T13) were processed for fluorescent immunohistochemistry and visualized by using confocal microscopy. Spinal pancreatic afferents were observed from T5 to T13, with the greatest contribution coming from T9-T12. The pancreatic afferents were equally distributed between right and left spinal ganglia; however, the innervation from the left NG was significantly greater than from the right. For both spinal and vagal afferents there was significantly greater innervation of the pancreatic head relative to the tail. The total number of retrogradely labeled afferents in the nodose was very similar to the total number of DRG afferents. The neurochemical phenotype of DRG neurons was dominated by transient receptor potential vanilloid 1 (TRPV1)-positive neurons (75%), GDNF family receptor alpha-3 (GFRalpha3)-positive neurons (67%), and calcitonin gene-related peptide (CGRP)-positive neurons(65%) neurons. In the NG, TRPV1-, GFRalpha3-, and CGRP-positive neurons constituted only 35%, 1%, and 15% of labeled afferents, respectively. The disparity in peptide and receptor expression between pancreatic afferents in the NG and DRG suggests that even though they contribute a similar number of primary afferents to the pancreas, these two populations may differ in regard to their nociceptive properties and growth factor dependency.

Role of vanilloid receptor 1 (VR1) in increased nociceptive signaling in experimental pancreatitis

Background/Aims: Vanilloid receptor 1 (VR1) plays a fundamental role in the transduction of pain due to peripheral tissue injury/inflammation. The aim of our study was to determine its role in visceral pain in a rat model of pancreetitis by measuring spinal cord c-fos as a functional marker. Methods: Pancreatitis was induced with two i.p. injections of L-arginine; control animals received saline and experiments were performed 5 days post-injection. Afferent responses were quantified by counting the number of c-fos immunoreactive neurons in spinal cord segments T8-TlO two hours after intraductal (i.d.) infusion of capsaicin (0.5 mg/kg), acid(pH 6) or vehicle with or without pre-treatment with the VR1 antagonist capsazepine (i.d.,O.5 mg/kg). To determine whether the number of pancreatic afferents expressing VR1 was increased in pancreatitis, the number of neurons in dorsal root ganglia T9-T11 positive for the retrograde label Dil and VR1 in pancreatitis and control rats was compared. Results: C-fos IR was significantly increased after acid or capsaicin stimulation compared to the vehicle injected group(40.9+-10.2/52.6+-11.8 vs 19.6+_6.6; p O.O5),capsazepine effectively blocked c-fos expression induced by acid and capsaicin(28.4+_8.6,p O.O5).Conciusions: The increased expression of c-los in response to capsaicin or acid stimulation in pancreatitis rats indicates that pancreatic afferents are sensitized in this model. Inhibition of the responses to acid and capsaicin by pre-treatment with a VR1 antagonist suggests that these responses require VR1 activation and that sensitization of afferent neurons in pancreatitis involves an increase in VR1 activity. These results imply that VR1 plays a critical role in pain transmission in pancreatitis. Our results also suggest that capsazepine may be an effective therapeutic agent in treating pain in pancreetitis.

Afferent innervation of the rat pancreas: retrograde tracing and immunohistochemistry in the dorsal root ganglia.

This study was undertaken to determine the segmental organization of the dorsal root ganglion (DRG) cells that give rise to pancreatic afferents containing a certain neuropeptide in the rat. These cells were examined using retrograde tracing combined with immunohistochemistry. Injection of horseradish peroxidase (HRP) into the pancreas resulted in the labeling of cells in bilateral T5-L2 DRGs, with most labeled cells lying at T10-T11. Injection into the duodenal (right), splenic (left), and entire lobes consistently produced more labeled cells significantly in the right, left, and right DRGs, respectively. Calcitonin gene-related peptide (CGRP)-, substance P (SP)-, somatostatin (SOM)-, and galanin (GAL)-immunoreactive (IR) cells in the DRGs (T9-T12) were found in -52, 17, 8, and 6%, respectively, but neuropeptide Y- and vasoactive intestinal polypeptide-IR cells were not found. About 88% of HRP-labeled cells in DRGs (T9-T12) contained CGRP, and approximately 16% of them contained SP. Although SOM- and GAL-IR cells were localized in the DRGs, these cells innervating the pancreas could not be found. In brief, these results show that bilateral (not similar in cell number on each side) DRG cells innervate the duodenal or splenic pancreas, and the majority of these cells that project to the pancreas contain CGRP and SP.

Role of pancreatic vagal and sympathetic afferents in the reflex increase of renal sympathetic activity after injection of bile into the canine pancreatic duct

Role of pancreatic vagal and sympathetic afferents in the reflex increase of renal sympathetic activity after injection of bile into the canine pancreatic duct

Cardiovascular reflexes arising from the gallbladder and pancreas in spinal and in decerebrate cats

Cardiovascular responses to chemical stimulation of thin-fiber afferents from the gallbladder and pancreas were determined before and after C 1 transection of the spinal cord in cats. Additional cats were studied before and after decerebration. Stimulation of gallbladder and pancreatic afferents caused significant increases in arterial pressure and heart rate in all groups; however, smaller responses often occurred in spinal cats. These results demonstrate that spinal circuitry alone can generate cardiovascular responses to visceral stimulation and that supraspinal, though not necessarily suprapontine areas, are involved in producing the full response.

Stimulation of pancreatic afferents reflexly activates the cardiovascular system in cats.

Chemical stimulation of afferents from the stomach and gallbladder has been shown reflexly to activate the cardiovascular system. It is not known, however, whether stimulating afferents from the pancreas evoke similar reflex activity. Therefore we recorded the cardiovascular responses in cats anesthetized with methoxyflurane, while we applied capsaicin (200 micrograms/ml) and bradykinin (0.001-1,000 micrograms/ml) to the surface of the pancreas. Topically applying these algesic substances evoked cardiovascular responses that included increases in systemic arterial pressure, heart rate, left ventricular dP/dt at 40-mmHg developed pressure and systemic vascular resistance. Bilateral vagotomy at the level of the diaphragm did not diminish the cardiovascular responses evoked by capsaicin or bradykinin. In contrast, removal of the celiac and superior mesenteric ganglia abolished the cardiovascular responses demonstrated previously when capsaicin or bradykinin was applied to the pancreas. We conclude that afferent endings in the pancreas can be stimulated reflexly to increase cardiovascular function in cats. This reflex activation represents a potential mechanism for eliciting the cardiovascular changes observed during acute pancreatitis, particularly the marked vasoconstriction that may lead to renal failure.

Visceral afferents and metabolic function.

The effect of glucose, 2 DG, CCK and serotonin on the firing rate of the vagal hepatic afferents and vagal pancreatic afferents was studied in rabbits, guinea pigs and rats. The hepatic and pancreatic afferents showed a decrease in activity following administration of glucose and an increase in activity after the injection of 2 DG. These afferent elements may have a mechanism to respond to blood glucose similar to that of lateral hypothalamic neurons. The results also indicate that CCK and serotonin showed the opposite effects in the vagal hepatic afferents and in the pancreatic efferents. Decrease in the discharge rate in the vagal hepatic afferents due to CCK or serotonin may cause a suppression of food intake behavior. Increase in the discharge rate in the vagal pancreatic afferents resulting from CCK or serotonin or insulin suggests the existence of a nervous information system for these substances in the circulating blood in the pancreas.