Hironobu Morita

Vestibular system plays a significant role in arterial pressure control during head-up tilt in young subjects.

We recently demonstrated that galvanic vestibular stimulation (GVS) obscured the vestibulo-cardiovascular reflex in rats during gravitational change. Here, we used GVS to examine the role of the vestibular-mediated arterial pressure (AP) control during a 60 degrees head-up tilt (HUT) in young and aged subjects (19-24 years old, 12 males and 3 females for young subjects, 63-91 years old, 5 males and 5 females for aged subjects). In young subjects, the AP did not change during posture transition from the supine position to HUT when GVS was not applied. When GVS was applied, AP immediately and significantly decreased by 17+/-2 mmHg upon HUT. When they were exposed to lower-body negative pressure (LBNP) in the supine position, the degree of footward fluid shift induced was the same as that induced by HUT. LBNP elicited only a footward fluid shift without alteration of vestibular input, while HUT elicited both. LBNP decreased the AP significantly, and the decrease was similar to that observed in the HUT with GVS. Thus, GVS modified the AP responses in young subjects during HUT but not during LBNP. In contrast, in aged subjects, the AP decreased during HUT regardless of whether GVS was applied. The vestibular system plays an important role in initial AP control during posture transition in young subjects. However, this function might be impaired in aged subjects.

A novel type of implantable and programmable infusion pump for small laboratory animals.

INTRODUCTION The iPRECIO (Primetech Corporation, Tokyo, Japan) is a new form of pump for infusing small laboratory animals. The key features of the iPRECIO are that it can be implanted within the animal, it is refillable, and it is programmable. The infusion start-points and end-points are adjustable, infusion rate can be altered, and the infusion solution can be changed after the pump is implanted. In order to confirm the precision of the iPRECIO, in vivo and in vitro experiments were employed. METHODS In the in vitro experiment, at the excretion rate of 1 microl/h for 336 h, 15 microl/h for 96 h, and 30 microl/h for 120 h, the decrease in each pump weight was used to estimate the actual excretion volume. In the in vivo experiments, the iPRECIO was chronically implanted in rats, angiotensin II was infused, and arterial pressure (AP) was monitored. RESULTS In the in vitro experiment, the volume of solution excreted from the pump increased with time, and the volume excreted matched the programmed volume. The infusion rate also changed at the scheduled time. In the in vivo experiment, AP increased and decreased on schedule, and a dose-dependent pressor response to angiotensin II occurred. Furthermore, after exchanging saline with angiotensin II, AP increased and decreased on schedule. DISCUSSION Present data of the in vitro and in vivo experiments indicates that the iPRECIO worked precisely, making it suitable for a variety of experiments involving small laboratory animals.

Interaction between vestibulo-cardiovascular reflex and arterial baroreflex during postural change in rats

To examine a cooperative role for the baroreflex and the vestibular system in controlling arterial pressure (AP) during voluntary postural change, AP was measured in freely moving conscious rats, with or without sinoaortic baroreceptor denervation (SAD) and/or peripheral vestibular lesion (VL). Voluntary rear-up induced a slight decrease in AP (-5.6 ± 0.8 mmHg), which was significantly augmented by SAD (-14.7 ± 1.0 mmHg) and further augmented by a combination of VL and SAD (-21 ± 1.0 mmHg). Thus we hypothesized that the vestibular system sensitizes the baroreflex during postural change. To test this hypothesis, open-loop baroreflex analysis was conducted on anesthetized sham-treated and VL rats. The isolated carotid sinus pressure was increased stepwise from 60 to 180 mmHg while rats were placed horizontal prone or in a 60° head-up tilt (HUT) position. HUT shifted the carotid sinus pressure-sympathetic nerve activity (SNA) relationship (neural arc) to a higher SNA, shifted the SNA-AP relationship (peripheral arc) to a lower AP, and, consequently, moved the operating point to a higher SNA while maintaining AP (from 113 ± 5 to 114 ± 5 mmHg). The HUT-induced neural arc shift was completely abolished in VL rats, whereas the peripheral arc shifted to a lower AP and the operating point moved to a lower AP (from 116 ± 3 to 84 ± 5 mmHg). These results indicate that the vestibular system elicits sympathoexcitation, shifting the baroreflex neural arc to a higher SNA and maintaining AP during HUT.

Impairment of vestibular-mediated cardiovascular response and motor coordination in rats born and reared under hypergravity.

It is well known that environmental stimulation is important for the proper development of sensory function. The vestibular system senses gravitational acceleration and then alters cardiovascular and motor functions through reflex pathways. The development of vestibular-mediated cardiovascular and motor functions may depend on the gravitational environment present at birth and during subsequent growth. To examine this hypothesis, arterial pressure (AP) and renal sympathetic nerve activity (RSNA) were monitored during horizontal linear acceleration and performance in a motor coordination task in rats born and reared in 1-G or 2-G environments. Linear acceleration of +/-1 G increased AP and RSNA. These responses were attenuated in rats with a vestibular lesion, suggesting that the vestibular system mediated AP and RSNA responses. These responses were also attenuated in rats born in a 2-G environment. AP and RSNA responses were partially restored in these rats when the hypergravity load was removed, and the rats were maintained in a 1-G environment for 1 wk. The AP response to compressed air, which is mediated independently of the vestibular system, did not change in the 2-G environment. Motor coordination was also impaired in the 2-G environment and remained impaired even after 1 wk of unloading. These results indicate that hypergravity impaired both the vestibulo-cardiovascular reflex and motor coordination. The vestibulo-cardiovascular reflex was only impaired temporarily and partially recovered following 1 wk of unloading. In contrast, motor coordination did not return to normal in response to unloading.

Vestibular-mediated increase in central serotonin plays an important role in hypergravity-induced hypophagia in rats

Exposure to a hypergravity environment induces acute transient hypophagia, which is partially restored by a vestibular lesion (VL), suggesting that the vestibular system is involved in the afferent pathway of hypergravity-induced hypophagia. When rats were placed in a 3-G environment for 14 days, Fos-containing cells increased in the paraventricular hypothalamic nucleus, the central nucleus of the amygdala, the medial vestibular nucleus, the raphe nucleus, the nucleus of the solitary tract, and the area postrema. The increase in Fos expression was completely abolished or significantly suppressed by VL. Therefore, these regions may be critical for the initiation and integration of hypophagia. Because the vestibular nucleus contains serotonergic neurons and because serotonin (5-HT) is a key neurotransmitter in hypophagia, with possible involvement in motion sickness, we hypothesized that central 5-HT increases during hypergravity and induces hypophagia. To examine this proposition, the 5-HT concentrations in the cerebrospinal fluid were measured when rats were reared in a 3-G environment for 14 days. The 5-HT concentrations increased in the hypergravity environment, and these increases were completely abolished in rats with VL. Furthermore, a 5-HT(2A) antagonist (ketanserin) significantly reduced 3-G (120 min) load-induced Fos expression in the medial vestibular nucleus, and chronically administered ketanserin ameliorated hypergravity-induced hypophagia. These results indicate that hypergravity induces an increase in central 5-HT via the vestibular input and that this increase plays a significant role in hypergravity-induced hypophagia. The 5-HT(2A) receptor is involved in the signal transduction of hypergravity stress in the vestibular nucleus.

Feasibility of a Short-Arm Centrifuge for Mouse Hypergravity Experiments.

To elucidate the pure impact of microgravity on small mammals despite uncontrolled factors that exist in the International Space Station, it is necessary to construct a 1 g environment in space. The Japan Aerospace Exploration Agency has developed a novel mouse habitat cage unit that can be installed in the Cell Biology Experiment Facility in the Kibo module of the International Space Station. The Cell Biology Experiment Facility has a short-arm centrifuge to produce artificial 1 g gravity in space for mouse experiments. However, the gravitational gradient formed inside the rearing cage is larger when the radius of gyration is shorter; this may have some impact on mice. Accordingly, biological responses to hypergravity induced by a short-arm centrifuge were examined and compared with those induced by a long-arm centrifuge. Hypergravity induced a significant Fos expression in the central nervous system, a suppression of body mass growth, an acute and transient reduction in food intake, and impaired vestibulomotor coordination. There was no difference in these responses between mice raised in a short-arm centrifuge and those in a long-arm centrifuge. These results demonstrate the feasibility of using a short-arm centrifuge for mouse experiments.

Long-term exposure to microgravity impairs vestibulo-cardiovascular reflex

The vestibular system is known to have an important role in controlling blood pressure upon posture transition (vestibulo-cardiovascular reflex, VCR). However, under a different gravitational environment, the sensitivity of the vestibular system may be altered. Thus, the VCR may become less sensitive after spaceflight because of orthostatic intolerance potentially induced by long-term exposure to microgravity. To test this hypothesis in humans, we investigated the ability of the VCR to maintain blood pressure upon head-up tilt before and after a 4–6 months stay on the International Space Station. To detect the functional state of the VCR, galvanic vestibular stimulation (GVS) was applied. As GVS transiently interrupts the vestibular-mediated pressor response, impaired VCR is detected when the head-up tilt-induced blood pressure response does not depend on GVS. During the first 20 s of head-up tilt, a transient blood pressure increase (11.9 ± 1.6 mmHg) was observed at pre-spaceflight but not at 1–4 days after return from spaceflight. The magnitude of VCR recovered to the pre-spaceflight levels within 2 months after return. These results indicate that long-term exposure to microgravity induces VCR impairment, which may be involved in a mechanism of spaceflight-induced orthostatic intolerance.

The vestibular system is critical for the changes in muscle and bone induced by hypergravity in mice

Gravity changes concurrently affect muscle and bone as well as induce alterations in vestibular signals. However, the role of vestibular signals in the changes in muscle and bone induced by gravity changes remains unknown. We therefore investigated the effects of vestibular lesions (VL) on the changes in muscle and bone induced by 3 g hypergravity for 4 weeks in C57BL/6J mice. Quantitative computed tomography analysis revealed that hypergravity increased muscle mass surrounding the tibia and trabecular bone mineral content, adjusting for body weight in mice. Hypergravity did not affect cortical bone and fat masses surrounding the tibia. Vestibular lesions blunted the increases in muscle and bone masses induced by hypergravity. Histological analysis showed that hypergravity elevated the cross‐sectional area of myofiber in the soleus muscle. The mRNA levels of myogenic genes such as MyoD, Myf6, and myogenin in the soleus muscle were elevated in mice exposed to hypergravity. Vestibular lesions attenuated myofiber size and the mRNA levels of myogenic differentiation markers enhanced by hypergravity in the soleus muscle. Propranolol, a β‐blocker, antagonized the changes in muscle induced by hypergravity. In conclusion, this study is the first to demonstrate that gravity changes affect muscle and bone through vestibular signals and subsequent sympathetic outflow in mice.

Arterial pressure oscillation and muscle sympathetic nerve activity after 20 days of head-down bed rest

Both spectral power within the low-frequency component, i.e., 0.04 to 0.15 Hz, of systolic pressure and muscle sympathetic nerve activity are increased during head-up tilt. The nerve activity during tilt is altered after space flight and exposure to simulated microgravity. In the present study, correlations of the low-frequency component and the nerve activity were analyzed before and after 20 days of -6° of head-down bed rest. Measurements were performed at -6° head-down bed rest, 0° (flat), and 30° and 60° head-up tilt (HUT). Mean arterial pressure during HUT was not different between pre- and post-bed rest, but muscle sympathetic nerve activity in post-bed rest significantly increased at tilt angles of -6°, 0°, 30°, and 60° compared with those during pre-bed rest. The low-frequency component of systolic pressure also significantly increased during post-bed rest compared with pre-bed rest at tilts of 0°, 30°, and 60°. The nerve activity and the frequency component were linearly correlated for individual (r(2) = 0.51-0.88) and averaged (r(2) = 0.60) values when the values included both pre- and post-bed rest. Thus, the low-frequency component of systolic pressure could be an index of the muscle sympathetic nerve activity during tilt during pre- and post-bed rest.

Galvanic vestibular stimulation counteracts hypergravity-induced plastic alteration of vestibulo-cardiovascular reflex in rats

Recent data from our laboratory demonstrated that, when rats are raised in a hypergravity environment, the sensitivity of the vestibulo-cardiovascular reflex decreases. In a hypergravity environment, static input to the vestibular system is increased; however, because of decreased daily activity, phasic input to the vestibular system may decrease. This decrease may induce use-dependent plasticity of the vestibulo-cardiovascular reflex. Accordingly, we hypothesized that galvanic vestibular stimulation (GVS) may compensate the decrease in phasic input to the vestibular system, thereby preserving the vestibulo-cardiovascular reflex. To examine this hypothesis, we measured horizontal and vertical movements of rats under 1-G or 3-G environments as an index of the phasic input to the vestibular system. We then raised rats in a 3-G environment with or without GVS for 6 days and measured the pressor response to linear acceleration to examine the sensitivity of the vestibulo-cardiovascular reflex. The horizontal and vertical movement of 3-G rats was significantly less than that of 1-G rats. The pressor response to forward acceleration was also significantly lower in 3-G rats (23 +/- 1 mmHg in 1-G rats vs. 12 +/- 1 mmHg in 3-G rats). The pressor response was preserved in 3-G rats with GVS (20 +/- 1 mmHg). GVS stimulated Fos expression in the medial vestibular nucleus. These results suggest that GVS stimulated vestibular primary neurons and prevent hypergravity-induced decrease in sensitivity of the vestibulo-cardiovascular reflex.