Claudine L. Frisby

Effects of insulin-like growth factor-I administration on radiation enteritis in rats.

BACKGROUND Acute radiation-induced damage to the small bowel occurs frequently during abdominal radiotherapy. Since the small intestine is selectively responsive to the growth-promoting effects of insulin-like growth factor-I (IGF-I), we investigated the effects of IGF-I administration on mucosal recovery from radiation enteritis in the rat. METHODS Rats received a single 10-Gy dose of total abdominal irradiation followed by implantation of mini-pumps infusing either IGF-I or vehicle for 4 days. After the rats had been killed, gut organs were weighed before light microscopic and biochemical examination. RESULTS Irradiated rats receiving IGF-I lost less body weight than vehicle-treated rats, whereas the wet weights of the stomach, small intestine, and colon were increased by 10%, 19%, and 21%, respectively, and crypt depth was increased in the duodenum, jejunum, and ileum. CONCLUSIONS IGF-I administration after abdominal irradiation increased small-intestinal mass and improved indicators of mucosal integrity, suggesting acceleration of small-intestinal mucosal recovery from radiation injury.

Sensory and Motor Innervation of the Crural Diaphragm by the Vagus Nerves

BACKGROUND & AIMS During gastroesophageal reflux, transient lower esophageal sphincter relaxation and crural diaphragm (CD) inhibition occur concomitantly. Modifying vagus nerve control of transient lower esophageal sphincter relaxation is a major focus of development of therapeutics for gastroesophageal reflux disease, but neural mechanisms that coordinate the CD are poorly understood. METHODS Nerve tracing and immunolabeling were used to assess innervation of the diaphragm and lower esophageal sphincter in ferrets. Mechanosensory responses of vagal afferents in the CD and electromyography responses of the CD were recorded in novel in vitro preparations and in vivo. RESULTS Retrograde tracing revealed a unique population of vagal CD sensory neurons in nodose ganglia and CD motor neurons in brainstem vagal nuclei. Anterograde tracing revealed specialized vagal endings in the CD and phrenoesophageal ligament-sites of vagal afferent mechanosensitivity recorded in vitro. Spontaneous electromyography activity persisted in the CD following bilateral phrenicotomy in vivo, while vagus nerve stimulation evoked electromyography responses in the CD in vitro and in vivo. CONCLUSIONS We conclude that vagal sensory and motor neurons functionally innervate the CD and phrenoesophageal ligament. CD vagal afferents show mechanosensitivity to distortion of the gastroesophageal junction, while vagal motor neurons innervate both CD and distal esophagus and may represent a common substrate for motor control of the reflux barrier.

Altered gastric vagal mechanosensitivity in diet-induced obesity persists on return to normal chow and is accompanied by increased food intake

Background and aims:Gastric vagal afferents convey satiety signals in response to mechanical stimuli. The sensitivity of these afferents is decreased in diet-induced obesity. Leptin, secreted from gastric epithelial cells, potentiates the response of vagal afferents to mechanical stimuli in lean mice, but has an inhibitory effect in high-fat diet (HFD)-induced obese mice. We sought to determine whether changes in vagal afferent function and response to leptin in obesity were reversible by returning obese mice consuming a HFD to standard laboratory chow diet (SLD).Methods:Eight-week-old female C57BL/6 mice were either fed a SLD (N=20) or HFD (N=20) for 24 weeks. A third group was fed a HFD for 12 weeks and then a SLD for a further 12 weeks (RFD, N=18). An in vitro gastro-oesophageal vagal afferent preparation was used to determine the mechanosensitivity of gastric vagal afferents and the modulatory effect of leptin (0.1–10 nM) was examined. Retrograde tracing and quantitative RT–PCR were used to determine the expression of leptin receptor (LepR) messenger RNA (mRNA) in whole nodose and specific cell bodies traced from the stomach.Results:After 24 weeks, both the HFD and RFD mice had increased body weight, gonadal fat mass, plasma leptin, plasma insulin and daily energy consumption compared with the SLD mice. The HFD and RFD mice had reduced tension receptor mechanosensitivity and leptin further inhibited responses to tension in HFD, RFD but not SLD mice. Mucosal receptors from both the SLD and RFD mice were potentiated by leptin, an effect not seen in HFD mice. LepR expression was unchanged in the whole nodose, but was reduced in the mucosal afferents of the HFD and RFD mice.Conclusion:Disruption of gastric vagal afferent function by HFD-induced obesity is only partially reversible by dietary change, which provides a potential mechanism preventing maintenance of weight loss.

Circadian Variation in Gastric Vagal Afferent Mechanosensitivity

Food intake is coordinated to cellular metabolism by clock gene expression with a master clock in the suprachiasmatic nucleus synchronized by light exposure. Gastric vagal afferents play a role in regulating food intake, but it is unknown whether they exhibit circadian variation in their mechanosensitivity. We aimed to determine whether gastric vagal afferents express clock genes and whether their response to mechanical stimuli oscillates throughout the light/dark cycle. Nodose ganglia were collected from 8-week-old female C57BL/6 mice every 3 h starting at lights off (1800 h) to quantify Bmal1, Per1, Per2, and Nr1d1 mRNA by qRT-PCR. Additionally in vitro single-fiber recordings of gastric vagal mechanoreceptors were taken at all time points. Per1, Per2, Bmal1, and Nr1d1 mRNA is expressed in the nodose ganglia and levels oscillated over a 24 h period. In mice fed ad libitum, gastric content was 3 times higher at 0000 h and 0300 h than 1200 h. The response of tension receptors to 3 g stretch was reduced by up to 70% at 2100 h, 0000 h, and 0300 h compared with 1200 h. Gastric mucosal receptor response to stroking with a 50 mg von Frey hair was 3 times greater at 1200 h and 1500 h than the response at 0000 h. Similar findings were obtained in mice fasted for 6 h or maintained in darkness for 3 d before study. Therefore, these changes do not result from food intake or the light/dark cycle. Thus, gastric vagal mechanoreceptors display circadian rhythm, which may act to control food intake differentially at different times of the day.

Effects of Fractionated Abdominal Irradiation on Small Intestinal Motility Studies in a Novel In Vitro Animal Model

Disordered small intestinal motility occurs frequently during acute radiation enteritis. However, the characteristics and time course of the motor dysfunction are poorly defined. These parameters were assessed in a novel animal model of radiation enteritis. Ileal pressures were recorded in vitro with perfused micromanometric catheter using an arterially perfused ileal loop in 22 ferrets following fractionated abdominal irradiation (9 doses 2.50 Gy thrice weekly for 3 weeks). Tissue damage was graded histologically. Studies were performed 3 to 29 days after irradiation. Tissue from 7 control animals was also studied. All treated animals developed diarrhoea. Histology showed changes consistent with mild to moderate radiation enteritis. Following irradiation, there was an initial increase in frequency followed by a non-significant reduction in the frequency, but not the amplitude of ileal pressure waves. The frequency of pressure waves showed an inverse relationship with time after radiation (r = -0.634, p < 0.002). There was no relationship between motility and histology. We conclude that abdominal irradiation is associated with a time-dependent reduction in ileal motility which does not correlate with light microscopic changes.

TRPV1 Channels and Gastric Vagal Afferent Signalling in Lean and High Fat Diet Induced Obese Mice

Aim Within the gastrointestinal tract vagal afferents play a role in control of food intake and satiety signalling. Activation of mechanosensitive gastric vagal afferents induces satiety. However, gastric vagal afferent responses to mechanical stretch are reduced in high fat diet mice. Transient receptor potential vanilloid 1 channels (TRPV1) are expressed in vagal afferents and knockout of TRPV1 reduces gastro-oesophageal vagal afferent responses to stretch. We aimed to determine the role of TRPV1 on gastric vagal afferent mechanosensitivity and food intake in lean and HFD-induced obese mice. Methods TRPV1+/+ and -/- mice were fed either a standard laboratory diet or high fat diet for 20wks. Gastric emptying of a solid meal and gastric vagal afferent mechanosensitivity was determined. Results Gastric emptying was delayed in high fat diet mice but there was no difference between TRPV1+/+ and -/- mice on either diet. TRPV1 mRNA expression in whole nodose ganglia of TRPV1+/+ mice was similar in both dietary groups. The TRPV1 agonist N-oleoyldopamine potentiated the response of tension receptors in standard laboratory diet but not high fat diet mice. Food intake was greater in the standard laboratory diet TRPV1-/- compared to TRPV1+/+ mice. This was associated with reduced response of tension receptors to stretch in standard laboratory diet TRPV1-/- mice. Tension receptor responses to stretch were decreased in high fat diet compared to standard laboratory diet TRPV1+/+ mice; an effect not observed in TRPV1-/- mice. Disruption of TRPV1 had no effect on the response of mucosal receptors to mucosal stroking in mice on either diet. Conclusion TRPV1 channels selectively modulate gastric vagal afferent tension receptor mechanosensitivity and may mediate the reduction in gastric vagal afferent mechanosensitivity in high fat diet-induced obesity.

Gastric neuropeptide W is regulated by meal-related nutrients

Neuropeptide W (NPW) is secreted from gastrin (G) cells in the stomach in response to food intake. The mechanisms underlying food intake-induced regulation of gastric NPW is largely unknown. We hypothesized that specific macronutrients were responsible for food-induced NPW secretion. We evaluated the acute effects of fat, carbohydrate and protein on plasma NPW concentrations in humans and mice. The effect of different nutrients on expression of NPW in the antral stomach was also determined in mice. Primary cell cultures of mouse gastric antral mucosal cells were used to investigate the signaling pathway of NPW expression. Plasma NPW concentrations did not change after nutrient ingestion in either humans or mice. NPW mRNA expression and the number of NPW positive cells in the mouse antrum were increased in mice gavage fed with protein or glucose, but not lipid. In primary antral mucosal cell culture, NPW mRNA expression was stimulated by l-phenylalanine, but not glucose. Calcium-sensing receptor (CaSR) positive cells were largely co-localized with NPW in mouse gastric antral mucosal cells, and NPW mRNA expression was inhibited by a selective antagonist of CaSR NPS2143. However, the l-phenylalanine-induced increase in NPW expression was not affected by NPS2143. In conclusion, these studies indicated an inconsistency between plasma and gastric NPW expression in response to nutrient ingestion, suggesting food induced gastric NPW expression may play a more important role locally. Moreover, glucose and especially protein are potent regulators of gastric NPW, via distinct mechanisms.

Neuropeptide W modulation of gastric vagal afferent mechanosensitivity: Impact of age and sex

AIM Gastric vagal afferents are activated in response to mechanical stimulation, an effect attenuated by neuropeptide W (NPW) in 20-week-old female mice. In this study we aimed to determine whether there were age and sex dependent effects of NPW on gastric vagal afferent mechanosensitivity. METHODS An in vitro gastro-oesophageal preparation was used to determine the effect of NPW on gastric vagal afferent mechanosensitivity from 8 and 20-week-old male and female C57BL/6 mice. Retrograde tracing and laser capture microdissection were used to selectively collect gastric vagal afferent cell bodies. Expression of NPW in the gastric mucosa and its receptor, GPR7, in gastric vagal afferent cell bodies was determined using quantitative RT-PCR. RESULTS NPW inhibited gastric tension sensitive vagal afferents from 20-week-old male and female mice, but not 8-week-old mice. In contrast, NPW inhibited the mechanosensitivity of gastric mucosal vagal afferents in 8-week-old male and female mice, but not 20-week-old mice. NPW mRNA expression in the gastric mucosa was higher in 20-week-old male mice compared to 8-week-old male mice. GPR7 mRNA expression in vagal afferent neurons innervating the gastric muscular layers was higher in 20-week-old mice compared to 8-week-old mice in both sexes. CONCLUSION The inhibitory effect of NPW on gastric tension sensitive and mucosal vagal afferents is age but not sex-dependent. These findings suggest that the physiological role of NPW varies depending on the age of the mice.

Divergence of mucosal and motor effects of insulin‐like growth factor (IGF)‐I and LR3IGF‐I on rat isolated ileum following abdominal irradiation

Abstract Background and Aims: In addition to its beneficial effects on small intestinal mucosal development and repair, insulin‐like growth factor (IGF)‐I has also been reported to improve neural function in toxic neuropathies. It has recently been recognized that enteric neural abnormalities contribute to the small intestinal dysmotility observed during and after abdominal radiotherapy for gynecological and pelvic malignancy. The aim of the present study was to evaluate the effects of IGF‐I (5 mg/kg per day) and the more potent analog LR3IGF‐I (5 mg/kg per day) on neurally mediated ileal dysmotility following irradiation.

Nesfatin-1 modulates murine gastric vagal afferent mechanosensitivity in a nutritional state dependent manner

HighlightsNesfatin‐1 potentiated mucosal receptor mechanosensitivity in lean fed and fasted but not high fat diet (HFD) mice.Tension receptor mechanosensitivity was unaffected by nesfatin‐1 in lean fed or fasted mice, but inhibited in HFD mice.Plasma concentrations of nesfatin‐1were unchanged between fed, fasted and HFD mice.No difference in NUCB2 mRNA expression was observed in gastric mucosa or gonadal fat of fed, fasted and HFD mice.Nesfatin‐1 modulated gastric vagal afferents in a nutritional state dependent manner. Abstract Food intake is regulated by vagal afferent signals from the stomach. Nesfatin‐1 is an anorexigenic peptide produced within the gastrointestinal tract and has well defined central effects. We aimed to determine if nesfatin‐1 can modulate gastric vagal afferent signals in the periphery and further whether this is altered in different nutritional states. Female C57BL/6J mice were fed either a standard laboratory diet (SLD) or a high fat diet (HFD) for 12 weeks or fasted overnight. Plasma nucleobindin‐2 (NUCB2; nesfatin‐1 precursor)/nesfatin‐1 levels were assayed, the expression of NUCB2 in the gastric mucosa and adipose tissue was assessed using real‐time quantitative reverse‐transcription polymerase chain reaction. An in vitro preparation was used to determine the effect of nesfatin‐1 on gastric vagal afferent mechanosensitivity. HFD mice exhibited an increased body weight and adiposity. Plasma NUCB2/nesfatin‐1 levels were unchanged between any of the groups of mice. NUCB2 mRNA was detected in the gastric mucosa and gonadal fat of SLD, HFD and fasted mice with no difference in mRNA abundance between groups in either tissue. In SLD and fasted mice nesfatin‐1 potentiated mucosal receptor mechanosensitivity, an effect not observed in HFD mice. Tension receptor mechanosensitivity was unaffected by nesfatin‐1 in SLD and fasted mice, but was inhibited in HFD mice. In conclusion, Nesfatin‐1 modulates gastric vagal afferent mechanosensitivity in a nutritional state dependent manner.