Microinjection of l-glutamate into the nucleus ambiguus partially inhibits gastric motility through the NMDA receptor - nitric oxide pathway.

We have previously reported that both l-glutamate (l-Glu) and nitric oxide (NO) modulate gastric motility in the nucleus ambiguus (NA). The aim of this study is to explore the potential correlation between the l-Glu and NO. A latex balloon connected to a pressure transducer was inserted into the pylorus through the fundus of anesthetized male Wistar rats to continuously record changes in gastric smooth muscle contractile curves. Pretreatment with the NO-synthase inhibitor N-nitro-l-arginine methylester (l-NAME) did not completely abolish the inhibitory effect of l-Glu on gastric motility, but intravenous injection of the ganglionic blocker hexamethonium bromide (Hb) did. By using a specific N-methyl-d-aspartic acid (NMDA) receptor antagonist, we blocked the inhibitory effect of the NO-donor sodium nitroprusside (SNP) on gastric motility. These results suggest that microinjections of l-Glu into the NA inhibits gastric motility by activating the cholinergic preganglionic neurons, partially through the NMDA receptor - NO pathway.

Comparative study of c-Fos expression in rat dorsal vagal complex and nucleus ambiguus induced by different durations of restraint water-immersion stress.

Restraint water-immersion stress (RWIS) of rats induces vagally-mediated gastric dysfunction. The present work explored the effects of different durations of RWIS on neuronal activities of the dorsal vagal complex (DVC) and the nucleus ambiguous (NA) in rats. Male Wistar rats were exposed to RWIS for 0, 30, 60, 120, or 180 min. Then, a c-Fos immunoperoxidase technique was utilized to assess neuronal activation. Resumptively, c-Fos expression in DVC and NA peaked at 60 min of stress, subsequently decreased gradually with increasing durations of RWIS. Interestingly, the most intense c-Fos expression was observed in the dorsal motor nucleus of the vagus (DMV) during the stress, followed by NA, nucleus of solitary tract (NTS) and area postrema (AP). The peak of c-Fos expression in caudal DMV appeared at 120 min of the stress, slower than that in rostral and intermediate DMV. The c-Fos expression in intermediate and caudal NTS was significantly more intense than that in rostral NTS. These results indicate that the neuronal hyperactivity of DMV, NA, NTS and AP, the primary center that control gastric functions, especially DMV and NA, may play an important role in the disorders of gastric motility and secretion induced by RWIS.

Effects of nucleus ambiguus and dorsal motor nuclei of vagus on gastric H(+) and HCO(3)(-) secretion in rats.

AIM: To determine the effects of electrical stimulation of nucleus ambiguus (NA) and dorsal motor nuclei of vagus (DMV) on gastric acid and bicarbonate secretion in rats. METHODS: NA and DMV in rats were electrically stimulated. Pylorus ligation or esophagus perfusion was used to collect the gastric secretion. The titratable H(+) quantum, H(+) concentration, HCO(3)(-) secretion quantum were measured. RESULTS: Electrical stimulation of NA had no effects on the volume of gastric juice, titratable acidity and acid concentration, but elicited a pronounced increase in the total bicarbonate. However, electrical stimulation of DMV significantly increased the titratable acidity, the volume of gastric juice and the acid concentration. Similarly, electrical stimulation of either NA or DMV decreased the respiratory frequency and sinus bradycardia. CONCLUSION: NA in rats can not control the secretion of gastric acid but the secretion of bicarbonate in gastric juice, while DMV controls the secretion of gastric acid.

Effects of electrical stimulation at different locations in the central nucleus of amygdala on gastric motility and spike activity.

The aim of the study was to determine the effects of electrical stimulation of different locations in the central nucleus of amygdala (CNA) on gastric motility and spike activity in dorsal vagal complex. Gastric motility index (GMI) and firing rate (FR) of dorsal vagal complex neurons were measured in adult Wistar rats respectively. Neuronal spikes in dorsal vagal complex (DVC) were recorded extracellularly with single-barrel glass microelectrodes. Each type of responses elicited by electrical stimulation in medial (CEM) and lateral (CEL) subdivisions of CNA were recorded, respectively. GMI was significantly increased after stimulation of CEM (p<0.01), and significantly decreased in response to CEL stimulation (p<0.01). After stimulation of CEM, FR in medial nucleus of the solitary tract (mNST) decreased by 31.6 % (p<0.01) and that in dorsal motor nucleus of the vagus (DMNV) increased by 27.1 % (p<0.01). On the contrary, FR in mNST increased (p<0.01) and that in DMNV decreased in response to CEL stimulation (p<0.05). In conclusions, our findings indicated that different loci of CNA may mediate differential effects on gastric activity via changes in the firing of brainstem neurons controlling gut activity.

[Time domain and power spectrum of wide frequency band electrocardiogram in pigeons].

The wide frequency band ECG (WFB-ECG) was recorded in 33 (anesthetized) normal pigeons by the microprocessor ECG system (made in Nanjing University) with a wide-frequency response (0-1000 Hz), a high-speed sweep (up to 1401 mm/s) and a high sensitivity (up to 28 mm/mV). The recording methods for limb leads in the pigeon were the same as those in man, except that the needle electrodes (made by No.5 needles) were subcutaneously inserted in the bases of the wings and in the legs. We studied the features of time domain and power spectrum of pigeons WFB-ECG. It presents P, R, S and T waves, but no Q wave, basically similar to the results from Aves described by Sturkie. But there are still many characters that were not be recorded on the conventional ECG: (1) the main QRS complex is inverted and forms the type of rS or rSr , no Q wave in leads II, III, aVF, and the S-T segment is absent, which is different from that of humans. The T wave is upright in leads II, III, and aVF (except one), in agreement with that of man. But in lead aVR, the main QRS complex is upright and forms the type of Rs, and the T wave is inverted without any exception. There is a large notch on the upstroke of S wave without any exception. The amplitude of the notch is 0.413+/-0.133 mV and the duration is 9.733+/-1.291 ms in lead II. (2) The ratio of duration of P wave to P-R segment is about 0.8, lower than that of humans (1.0-1.6), but higher than that of mice (0.4). (3) The low frequency signals (0-80 Hz) are prominent. The relative power content of high frequency range of QRS in lead II is: 100-1000 Hz: (10.181+/-7.443)%; 80-300 Hz: (15.418+/-10.579)%. (4)The QRS vector loop in the frontal plane lies between -90 degrees and -180 degrees. The electrical axis of QRS complex averages -118 +/-10 (ranges from -96 degrees to -136 degrees). The reason that position of vector loop and the direction of main wave of QRS in the pigeon are different from human s and rodent s is probably that the Purkinje fibers cross the whole ventricular wall and terminate in the subepicardium in Aves including pigeons . After the impulses coming from the sinoatrial node reach the ventricular muscles, the subepicardium is depolarized before the endocardium. However in human s and rodent s, the Purkinje fibres only reach one-forth to one-second of the whole thickness from the endocardium to the epicardium, the subendocardium is depolarized before the subepicardium.

Oxytocin and vasopressin involved in restraint water-immersion stress mediated by oxytocin receptor and vasopressin 1b receptor in rat brain.

AIMS: Vasopressin (AVP) and oxytocin (OT) are considered to be related to gastric functions and the regulation of stress response. The present study was to study the role of vasopressinergic and oxytocinergic neurons during the restraint water-immersion stress. METHODS: Ten male Wistar rats were divided into two groups, control and RWIS for 1h. The brain sections were treated with a dual immunohistochemistry of Fos and oxytocin (OT) or vasopressin (AVP) or OT receptor or AVP 1b receptor (V(1b)R). RESULTS: (1) Fos-immunoreactive (Fos-IR) neurons dramatically increased in the hypothalamic paraventricular nucleus (PVN), the supraoptic nucleus (SON), the nucleus of solitary tract (NTS) and motor nucleus of the vagus (DMV) in the RWIS rats; (2) OT-immunoreactive (OT-IR) neurons were mainly observed in the medial magnocellular part of the PVN and the dorsal portion of the SON, while AVP-immunoreactive (AVP-IR) neurons mainly distributed in the magnocellular part of the PVN and the ventral portion of the SON. In the RWIS rats, Fos-IR neurons were identified in 31% of OT-IR neurons and 40% of AVP-IR neurons in the PVN, while in the SON it represented 28%, 53% respectively; (3) V(1b)R-IR and OTR-IR neurons occupied all portions of the NTS and DMV. In the RWIS rats, more than 10% of OTR-IR and V(1b)R-IR neurons were activated in the DMV, while lower ratio in the NTS. CONCLUSION: RWIS activates both oxytocinergic and vasopressinergic neurons in the PVN and SON, which may project to the NTS or DMV mediating the activity of the neurons by OTR and V(1b)R.

The role of catecholaminergic neurons in the hypothalamus and medullary visceral zone in response to restraint water-immersion stress in rats.

The activity of catecholaminergic neurons in the hypothalamus and the medullary visceral zone (MVZ) in rats in response to restraint water-immersion stress (RWIS) was measured by use of dual Fos and tyrosine hydroxylase (TH) immunohistochemistry. In RWIS rats Fos immunoreactive (Fos-IR) nuclei dramatically increased in the paraventricular nucleus (PVN), the supraoptic nucleus (SON), the dorsal motor nucleus of the vagus (DMV), the nucleus of the solitary tract (NTS), the area postrema (AP), and the ventrolateral medulla (VLM). A small number of TH-immunoreactive (TH-IR) and Fos/TH double-labeling neurons in the PVN, and their absence from the SON, were observed in both RWIS and nonstressed rats. More TH-IR neurons were observed in the MVZ of RWIS rats than in nonstressed rats. In RWIS and nonstressed rats, the percentage of Fos-IR nuclei in TH-IR neurons was 38.0 and 14.3% in the DMV, 34.4 and 9.7% in the NTS, 18.6 and 4.5% in the AP, and 45.7 and 18.9% in the VLM, respectively. In conclusion, catecholaminergic neurons in the MVZ are involved in the response to RWIS; although the PVN and SON also participate in the response to RWIS, the mechanism is not via catecholaminergic neurons.

Studies on functional connections between the supraoptic nucleus and the stomach in rats.

The present study was to investigate whether there are functional connections between the hypothalamic supraoptic nucleus (SON) and the stomach, which is the case with the paraventricular nucleus. The rats were divided into four groups. Group I: the neuronal discharge was recorded extracellularly in the NTS, DMV or SON before and after cold physiological saline (4°C) was perfused into the stomach and effused from the duodenum. Group II: the rats were stimulated as for Group I and c-Fos expression in NTS, DMV and SON was examined. Group III: the control to Group II. Group IV: gastric motility was recorded continuously before and after microinjection of L: -Glu into the SON. In Group I, the discharge frequency increased in all the three nuclei, while in Group II, Fos expression in NTS, DMV and SON was, respectively, greater than that of Group III. In Group IV, microinjection of L: -Glu (5 nmol) into SON significantly inhibited gastric motility. These data suggest there are functional connections between SON and stomach.

[Effects of L-glutamate microinjection into the ventral horn of spinal cord gray matter on gastrocnemius contractility in toad].

A diagram of motor neuron pool of ventral horn of spinal cord gray matter in toad was first delineated. Different concentrations (1, 0.5, 0.1, 0.01 mol/L) of excitatory amino acid L-Glu or physiological saline (0.65% NaCl) were then microinjected into the motor neuron pool in a urethane-anaesthetized toad. The contraction curve of the gastrocnemius was then recorded by the BL-420 Physiological Signal Recording. We took the maximal tension, the duration of rising phase, the velocity of tension variation, and the duration of descending phase as the parameters to study the characteristic of gastrocnemius contractility. It was found that the gastrocnemius contractility of all the 4 groups was tetanus but differed in degree, especially the maximal tension, and velocity of tension variation. In contrast to physiological saline, gastrocnemius contracted by the stimulation of L-Glu, and the contraction parameter showed dose-effect relationships except for the duration of descending phase, which was caused by the combination rate of L-Glu and the receptor.

Effects and mechanisms of L-glutamate microinjected into nucleus ambiguus on gastric motility in rats.

BACKGROUND: L-glutamate (L-GLU) is a major neurotransmitter in the nucleus ambiguus (NA), which can modulate respiration, arterial pressure, heart rate, etc. This study investigated the effects and mechanisms of L-GLU microinjected into NA on gastric motility in rats. METHODS: A latex balloon connected with a pressure transducer was inserted into the pylorus through the forestomach for continuous recording of the gastric motility. The total amplitude, total duration, and motility index of gastric contraction waves within 5 minutes before microinjection and after microinjection were measured. RESULTS: L-GLU (5 nmol, 10 nmol and 20 nmol in 50 nl normal saline (PS) respectively) microinjected into the right NA significantly inhibited gastric motility, while microinjection of physiological saline at the same position and the same volume did not change the gastric motility. The inhibitory effect was blocked by D-2-amino-5-phophonovalerate (D-AP5, 5 nmol, in 50 nl PS), the specific N-methyl-D-aspartic acid (NMDA) receptor antagonist, but was not influenced by 6-cyaon-7-nitroquinoxaline-2,3-(1H,4H)-dione (CNQX) (5 nmol, in 50 nl PS), the non-NMDA ionotropic receptor antagonist. Bilateral subdiaphragmatic vagotomy abolished the inhibitory effect by microinjection of L-GLU into NA. CONCLUSIONS: Microinjection of L-GLU into NA inhibits the gastric motility through specific NMDA receptor activity, not non-NMDA receptor activity, and the efferent pathway is the vagal nerves.

Hindbrain Effects of L-Glutamate on Gastric Motility in Rats.

BACKGROUND: There are no unanimous standpoints about the dorsal motor nucleus of the vagus (DMV) and nucleus of solitary tract (NTS) involving in the regulation of gastric motility up to now. METHODS: In this study, we injected L-Glutamate (L-Glu), an incitant neurotransmitter in the central neural system, into DMV and NTS to further investigate the effects of the two nuclei on gastric motility. A latex balloon connected with a pressure transducer was inserted into the pylorus through the fundus for continuous recording of the change of gastric smooth muscle contractile curves. RESULTS: L-Glu (10 nmol in 0.1 µl) microinjected into right DMV and NTS significantly inhibited gastric motility. We compared the effects of L-Glu (10 nmol) microinjected into the two nuclei, the L-Glu microinjected into right NTS had the greater inhibitory effect on gastric motility than microinjected into the right DMV. The physiological saline microinjection evoked no significant effect on gastric motility. CONCLUSIONS: L-Glu microinjected into right DMV and NTS evoked significant inhibition on gastric motility in rats. At equal dose of L-Glu, NTS had the greater inhibitory effect than DMV.

c-Fos expression in the supraoptic nucleus is the most intense during different durations of restraint water-immersion stress in the rat.

Restraint water-immersion stress (RWIS) can induce anxiety, hypothermia, and severe vagally-mediated gastric dysfunction. The present work explored the effects of different durations of RWIS on neuronal activities of the forebrain by c-Fos expression in conscious rats exposed to RWIS for 0, 30, 60, 120, or 180 min. The peak of c-Fos induction was distinct for different forebrain regions. The most intense c-Fos induction was always observed in the supraoptic nucleus (SON), and then in the hypothalamic paraventricular nucleus (PVN), posterior cortical amygdaloid nucleus (PCoA), central amygdaloid nucleus (CeA), and medial prefrontal cortex (mPFC). Moreover, body temperature was reduced to the lowest degree after 60 min of RWIS, and the gastric lesions tended to gradually worsen with the prolonging of RWIS duration. These data strongly suggest that these nuclei participate in the organismal response to RWIS to different degrees, and may be involved in the hypothermia and gastric lesions induced by RWIS.

[Studies on effects and mechanism of water immersion restraint stress on bile secretion in rats].

AIM: To probe into the operation mechanism of stress, through the studies on the effects of bile secretion in rats at the condition of water immersion restraint. METHODS: The animals were divided into six groups (n=8): Group A: restraint alone under room temperature + saline; Group B: water immersion restraint + saline; Group C: restraint alone under room temperature + Atropine; Group D: water immersion restraint + Atropine; Group E: restraint alone under room temperature + Phentolamine; F group: water immersion restraint + Phentolamine. RESULTS: Compared with group A, the capacity of bile secretion in group B decreased significantly (P < 0.05), changes of bile increased remarkably (P < 0.01), but there were no significant decreases on the capacity of bile secretion in group C (P > 0.05) compared with A, Group C only decreased appreciably. Compared with group A, the capacity of bile secretion in group E decreased appreciably (P < 0.05). Compared with group B, the capacity of bile secretion in group D decreased significantly (P < 0.05), pH of bile had no significant changes in group D. Compared with group B, the capacity of bile secretion in group F decreased significantly (P < 0.05), pH of bile had no significant changes in group F. Compared with group D, the capacity of bile secretion and pH of bile in group F had no significant changes. CONCLUSION: Water immersion restraint stress inhibited evidently on the capacity of bile secretion, and the capacity of bile secretion in water immersion groups decreased significantly, moreover pH of bile increased greatly. At the condition of restraint alone under room temperature, vagus and sympathetic nerve had no significant effects on the bile secretion, but they played important roles in decreases of bile secretion evidently induced by water immersion restraint stress in rats (P < 0.05).

Comparative studies on the wide frequency band electrocardiogram and vectorcardiogram in pigeon and mouse.

Wide frequency band ECG and vectorcardiogram in anesthetized pigeon and mouse were studied from the standpoint of comparison. The key results were as follows: in pigeon, the direction of the main QRS was inverted in leads II, III and aVF, and upright in lead aVR, which was contrary to that in mouse. The T wave was upright in leads II, III and aVF, but inverted in lead aVR in pigeon, which was the same as that in mouse. In pigeon, there was a large notch on the upstroke of the S wave in lead II without exception, but there was no such notch in the corresponding lead in mouse. The QRS vector loop in the frontal plane lay between -90 and -180 degrees in pigeon, while that of mouse lay between 0 and 90 degrees. The relative power of high frequency range (80-1000 Hz) of the QRS in lead II was approximately 15% in pigeon, but 55% in mouse. The direction of the main QRS was contrary in pigeon and mouse because the subepicardial muscles were depolarized before the subendocardial muscles in pigeon, but the latter were depolarized before the former in mouse. The direction of the T waves was the same in both pigeon and mouse because subepicardial muscles were all repolarized before subendocardial muscles.