Vinyl isolator breeding induces insulin resistance in C57BL/6JJcl mice.

As basic probiotics studies, the glucose tolerance test (GTT), insulin tolerance test (ITT), and adipokine and hepatic enzyme activities were investigated in male C57BL/6JJcl (B6J) mice under germfree (GF) or specific pathogen free (SPF) conditions. GF B6J mice were reproduced by reproductive engineering and cesarean section using a vinyl isolators (GF group). Some GF group mice were transferred to other vinyl isolators under SPF conditions (SPF group). In addition, conventional B6J mice bred in an open room were defined as controls (Conv group). GTT, ITT, and the sampling of blood, liver, white adipose tissue, and pancreas were performed when these B6J mice were at the age of 8 weeks. As a result, the GF and SPF groups showed hyperglycemia, impaired glucose tolerance and insulin resistance when compared with the Conv group. The adipose tissues and plasma TNFα concentrations in the GF and SPF groups were enlarged and increased when compared with the Conv group. Hepatic enzyme activities associated with glucose uptake in the GF and SPF groups were higher than those in the Conv group. However, hepatic enzyme activities associated with gluconeogenesis in the GF and SPF groups were lower than those in the Conv group. We assumed that these results were reactions by the liver to recover from the impaired glucose tolerance and the insulin resistance caused by vinyl isolator breeding of the GF and SPF groups by control of glucose metabolism.

Morphological and sequence analysis of Mycoplasma sp. isolated from the oral cavity of a house musk shrew (Suncus murinus).

Mycoplasma sp. strain EDS-4 was isolated from the oral cavity of EDS line of a house musk shrew (Suncus murinus) originated from Bangladesh, and was distinguished from all previously described mollicutes. It lacks a cell wall; ferments glucose; does not produce film and spots; and does not hydrolyse arginine and urea. The strain could be distinguished from all previously described mollicutes by 16S rRNA gene sequence comparisons. The results suggest that the isolate is new species of mollicutes originated from the shrew. The strain EDS-4 has been deposited with Japan collection of Microorganisms, Bioresource Center, RIKEN in Japan (JCM15930). The 16S rRNA gene sequence of strain EDS-4 is available through the DDBJ under accession number (AB469852).

Interactions of carotid sinus or aortic input with emetic signals from gastric afferents and vestibular system.

This study investigated how baro- and chemoreceptor afferents interact with emetic signals from gastric afferents and the vestibular system, and how these interactions modulate emetic and prodromal responses. We performed splanchnic denervation and abdominal vagotomy in anesthetized shrews (Suncus murinus), and then induced emetic responses by gastric distension. Next, we investigated the effects of these gastric afferent sections on cardiovascular and emetic responses induced by electrical stimulation of the aortic depressor nerve (ADN) and the carotid sinus nerve (CSN) with or without gastric distension. Splanchnic denervation abolished the prodromal response before retching and aortic baroreflex inhibition caused by gastric distension, but had no effects on the emetic response. In contrast, abdominal vagotomy abolished the emetic response induced by gastric distension with or without CSN stimulation, but without affecting gastric distension-induced or CSN stimulation-induced vascular and respiratory responses. In conscious animals, CSN denervation significantly suppressed veratrine- and motion-induced emetic responses, whereas ADN denervation had no significant effects. These results suggest that aortic baroreflex inhibition via the activation of splanchnic afferents contributes to the prodromal response before retching and circulatory homeostasis. In contrast, carotid sinus inputs, which are usually non-emetic signals, interact with vagal and vestibular inputs, and modulate the development of retching and vomiting.

Modulation of emetic response by carotid baro- and chemoreceptor activations.

We hypothesized that baroreceptor or chemoreceptor activation might be involved in the emetic, and prodromal cardiovascular and respiratory responses. To test this hypothesis, we induced the emetic responses by gastric distension in anesthetized Suncus murinus (house musk shrew), that had intact and absent baroreceptor and chemoreceptor afferents. Secondly, we stimulated the aortic depressor nerve (ADN) and the carotid sinus nerve (CSN) with or without gastric distension. Internal carotid artery ligation in the bifurcation area, which abolished reflex bradycardia by baroreceptor activation, and abolition of chemoreceptor reflex bradycardia and hyperventilation, by carotid body denervation, suppressed the emetic response but did not abolish it. ADN denervation, which produced no significant effects on the baroreceptor or chemoreceptor reflex bradycardia, had no effect on the emetic response, including the prodromal phase. CSN stimulation with gastric distension elicited retching accompanied by reflex bradycardia and hypotension during or just after stimulation, whereas ADN stimulation with gastric distension did not induce the cardiovascular reflex, and had no effects on the emetic response. These results indicate that carotid, rather than aortic, baroreceptor or chemoreceptor activation plays an important role in the augmentation of cardiac parasympathetic activity and the development of emetic response. In conclusion, carotid baroreceptor or chemoreceptor activation, which is non-emetic stimulation, acts as a modulator in the central mechanisms of emesis.

Emetic responses and neural activity in young musk shrews during the breast-feeding/weaning period: comparison between the high and low emetic response strains using a shaking stimulus.

The high emetic response (HER) strain and low emetic response (LER) strain of musk shrews (Suncus murinus) markedly differ in the emetic reflex in adults. However, there have been no studies on young musk shrews. We gave a shaking stimulus to young musk shrews aged 10 days or more that were obtained by mating within each strain and observed emetic responses. In the HER strain, no animal aged 10 days vomited, but vomiting was observed in 1 of 5 animals each aged 12 and 14 days, 2 of 5 animals aged 16 days, and all animals aged 18 days or more. In the LER strain, no vomiting was observed until the age of 14 days, but at the age of 16 days or more, 1 or 2 of 5 animals at each age vomited. After stimulation, activated neurons of the dorsal vagal complex and the dorsal reticular formation of the nucleus ambiguus (Amb) were examined by Fos immunohistochemistry. This morphometric study demonstrated that the numbers of Fos-positive neurons in the nucleus of the solitary tract and the dorsal reticular formation of the Amb were significantly larger in the animals that vomited in the HER strain than animals that did not vomit in the LER strain. We suggest that neurons in these regions are involved in emetic responses, as is the case in adult animals.

Induction of Fos protein in neurons in the medulla oblongata after motion- and X-irradiation-induced emesis in musk shrews (Suncus murinus).

To clarify the anatomical location of medullary neurons associated with vomiting, the musk shrew (Suncus murinus), a small animal used as a model for emesis, was exposed to various emetic stimuli and patterns of neuronal excitation were investigated by Fos immunohistochemistry. In motion experiments, musk shrews were shaken for 30 min on a tabletop shaker (displacement=25 mm and frequency=1.2 Hz). Ten of fifteen animals vomited frequently (Mo-FV group); the other five animals did not vomit (Mo-NV group). In radiation experiments, X-ray irradiation (10 Gy) of the whole body caused frequent vomiting in all of seven experimental animals (Ra-FV group). In the Mo-FV group, many Fos-immunoreactive (Fos-ir) neurons were detected in the nucleus of the solitary tract (NTS) and the reticular formation. The distribution pattern of Fos-ir neurons in the Mo-NV group was similar to that in the Mo-FV group, but the Mo-NV group had significantly fewer positive neurons in the NTS and the reticular formation around the nucleus ambiguus. In the Ra-FV group, numerous Fos-ir neurons were observed in the area postrema, an area containing no positive neurons in the motion-stimulated animals. The number of Fos-ir neurons in the NTS of the Ra-FV group was not statistically different from that of the Mo-NV group. In the Mo-FV and Ra-FV groups, Fos-ir neurons were clustered in the reticular formation at the dorsal-dorsomedial edge of the nucleus ambiguus at the level of the rostral medulla, while few such clusters were observed in the Mo-NV group. These neurons may play a role in the regulation of the vomiting response.

Role of the autonomic nervous system in emetic and cardiovascular responses in Suncus murinus.

To clarify the role of the autonomic nervous system in cardiovascular and emetic responses, we studied the influence of drugs that act on autonomic nervous function on emetic and cardiovascular responses induced by chemical or mechanical stimulation to the stomach in two strains of Suncus murinus, Jic:SUN-Her and Jic:SUN-Ler. Latency to the first retching in Jic:SUN-Her was significantly shorter than that in Jic:SUN-Ler to both mechanical and chemical stimulation. This result indicated that there are different sensitivities to mechanical and chemical stimulation to the stomach in these two strains of suncus. However, the numbers of emetic episodes were almost the same in these two strains. Mean blood pressure significantly increased from baseline prior to retching in both strains. Heart rate decreased in Jic:SUN-Her and increased in Jic:SUN-Ler prior to retching, suggesting that a different baroreflex responsiveness might exist in these two strains of suncus. Administration of acetylcholine and phenylephrine affected emetic response induced by mechanical and chemical stimulation. Although the baseline values of mean blood pressure and heart rate after administration of these drugs were different, changes in mean blood pressure and heart rate prior to retching were unaffected. This result suggested that the state of autonomic activity before the emetic response might be important in the development of the emetic response. Pretreatment with hexamethonium suppressed the cardiovascular response prior to retching and prolonged the latency to the first retching. This result indicated that there was an interaction between the mechanisms involved in cardiovascular and emetic responses. The change in autonomic function during the emetic response, especially enhancement of sympathetic activity prior to retching, may be relevant to emetic and cardiovascular responses. Moreover, these results suggest that different autonomic function or different baroreflex responsiveness in Jic:SUN-Her and Jic:SUN-Ler may be involved in emetic responses.

Immunohistochemical demonstration of c-fos protein in neurons of the medulla oblongata of the musk shrew (Suncus murinus) after veratrine administration.

We subcutaneously injected 0.5 mg/kg veratrine into the musk shrew (Suncus murinus), observed the presence or absence, latency, and the incidence of vomiting in each animal for 90 min, and selected animals that frequently vomited (FV group) and those that did not vomit (NV group). Subsequently, animal brains were removed, and the induction of c-fos protein (Fos) was immunohistochemically examined to evaluate neuronal activity in the medulla oblongata. The distribution of Fos-positive neurons in the medulla oblongata was similar between FV and NV groups, with numerous neurons along the entire length of the nucleus of the solitary tract and in the ventrolateral reticular formation. Both veratrine-injected groups showed higher numbers of positive neurons than the saline administered group. However, while the FV group showed a high concentration of positive neurons in the dorsal-dorsomedial reticular formation of the nucleus ambiguus in the rostral medulla, the NV group showed few positive neurons in this area. Fos activity in neurons in this area appeared to be higher in animals with a higher incidence of vomiting.

Diurnal fluctuations of heart rate, body temperature and locomotor activity in the house musk shrew (Suncus murinus).

Diurnal fluctuations of heart rate (HR), body temperature (BT) and locomotor activity (LA) in the unanaesthetized and unrestrained house musk shrew (Suncus murinus) were studied using a telemetry system. Six adult male shrews (Jic:SUN) weighing 60-70 g were used in the present study. They were housed under conditions of 24 C and a 12/12-hr light-dark cycle. HR, BT and LA were recorded over 10 days, following the post-implantation period (10 days or more) of the telemetric transmitter. A clear nocturnal rhythm of LA was shown, while intermittent and short-term LA were shown during the light period. The mean HR was 323.5 +/- 8.8 bpm in the light period and 354.3 +/- 5.2 bpm in the dark period, and the fluctuation of HR showed a nocturnal pattern. A nocturnal pattern was also observed in BT fluctuation, and all animals lowered their body temperature from 35-37 C to approximately 30 C or below, mostly during the light period. The fall of body temperature progressed over 2-3 hr, and then rose to the baseline temperature rapidly within approximately 30 min. While the body temperature fell, HR markedly decreased to approximately 100 bpm. These results suggest that the shrew has unique physiological properties in maintaining metabolic balance which are anticipated to be caused by the dramatic alteration of the autonomic nervous function.