We studied the effects of simulated microgravity on mouse oocytes maturation, and analyzed whether the tail-suspended model can be applied to investigate simulated microgravity effects on reproductive processes in female mice. Mouse oocytes were cultured in vitro with microgravity simulated by a rotating wall vessel bioreactor and by tail-suspended model, and the maturation rate of the mouse oocytes in the two models were examined in vivo. The maturation rate of mouse oocytes cultured in simulated microgravity was 8.93%, and that was 72.33% in 1g gravity. In ratio, oocyte maturation rate had no significant difference between the rotational group and control group. Microgravity simulated by the tail-suspended model inhibited mouse oocytes maturation and increased the rate of oocytes abnormity. The maturation rate of tail-suspended mouse oocytes was 14.54%, which was significantly lower than that of control group. Tail-suspended model should be an ideal model to investigate simulated microgravity effects on reproductive processes of female mice.
Animals ; Cells, Cultured ; Female ; Hindlimb Suspension ; Mice ; Oocytes ; cytology ; physiology ; Oogenesis ; physiology ; Weightlessness Simulation

The aim of the present study was to examine whether there are changes in systolic and diastolic blood pressure (SBP and DBP) and heart rate (HR) and their spectral indices in conscious free-moving rats after tail-suspension for 28 d. The tail-suspended hindlimb-unloaded (HU) rat model was used to simulate the cardiovascular effect of microgravity and the post-spaceflight cardiovascular dysfunction. The auto- and cross-spectral analysis of SBP variability (SBPV) and HR variability (HRV) were performed by the method based on the autoregressive model (AR), and the auto-spectral results was compared with the results from the classical periodogram method. The baroreceptor-heart rate reflex sensitivity (BRS) was estimated using transfer function analysis from SBP to HR. The results indicated that auto-spectral results based on the two methods were comparable, while smoother power spectral curves with distinguished peaks were trained by the AR method. The means of SBP, DBP, and HR, the main spectral indices of SBPV and HRV, and the mean average gain of transfer functions computed at low- and high-frequency ranges (0.25-0.8 Hz and 0.8-2.4 Hz) did not show significant changes before and after release from suspension. Furthermore, the main spectral indices of SBPV and HRV at different time points did not show significant differences between the control and suspension groups. However, the means of SBP, DBP, and HR at different time points were significantly higher in simulated weightless rats than those in the control rats. The findings of the present study suggest that a mid-term simulated microgravity might induce hypertension and tachycardia upon removal from the suspension which reflects a general sympathetic hyperactivity. We speculated that the sympathetic hyperactivity might be a compensatory mechanism activated in the intact animal to counteract HU-induced hypo-responsiveness of resistance vessels. In addition, lack of clear and distinct changes in HRV and BRS have also been reported in some recent space and ground-based human studies.
Animals ; Baroreflex ; Blood Pressure ; Heart Rate ; Hindlimb Suspension ; Hypertension ; Rats ; Weightlessness Simulation

OBJECTIVETo study the effects on rats' bone mineral density and bone biomechanics by suspensory simulated weightlessness and removing suspension.

METHODSTwenty Wistar rats were divided into two groups randomly as control group and model group. Suspend the model group rats for 14 days, then remove suspension and continue to feed for another 14 days. Feed control group rats for 28 days. Detect the bone mineral density (BMD) in vivo of cranial bone, second thoracic vertebra, fourth lumbar vertebra, pelvis, right radioulna and right femoral bone of each group at the 14th day. At the 28th day,execute all the rats and take out of right femoral bone and fourth lumbar vertebra for detecting BMD and the intensity of biomechanics.

RESULTSAt the 14th day in experiment, being compared with control group,the BMD of femoral bone, pelvis and lumbar vertebra in model group decreased significantly (P < 0.001, P < 0.001, P < 0.01) and the change of BMD of cranial bone, thoracic vertebra and radioulna in model group was not remarkable (P > 0.05). At the 28th day in experiment, the BMD of femoral bone and lumbar vertebra, the maximal load of femoral bone decreased significantly in model group as compared with control group (P < 0.01, P < 0.001, P < 0.01).

CONCLUSIONBMD in vive body showed that suspensory simulated weightlessness for 14 days could cause disorder of bone metabolism and remarkable mineral loss of weight bearing bone, even BMD and biomechanical intensity of weight bearing bone decrease obviously when removing suspension for 14 days. The results suggest that the disorder of bone metabolism could not be recovered in short time.

Animals ; Biomechanical Phenomena ; Bone Density ; Bone and Bones ; physiology ; Hindlimb Suspension ; Male ; Rats ; Rats, Wistar ; Weightlessness Simulation

One of the most important and urgent issues in the field of space medicine is to reveal the potential mechanism underlying the disused muscle atrophy during the weightlessness or microgravity environment. It will conduce to find out effective methods for the prevention and treatment of muscle atrophy during a long-term space flight. Increasing data show that muscle spindle discharges are significantly altered following the hindlimb unloading, suggesting a vital role in the progress of muscle atrophy. In the last decades, we have made a series of studies on changes in the morphological structure and function of muscle spindle following simulated weightlessness. This review will discuss our main results and related researches for understanding of muscle spindle activities during microgravity environment, which may provide a theoretic basis for effective prevention and treatment of muscle atrophy induced by weightlessness.
Animals ; Hindlimb Suspension ; Muscle Spindles ; physiopathology ; Muscle, Skeletal ; physiopathology ; Muscular Atrophy ; physiopathology ; Space Flight ; Weightlessness Simulation

OBJECTIVE: To introduce a modified protocol for generating the simulated weightlessness rat model by hindlimb unloading. METHODS: Ninety male adult SD rats were randomly divided into three groups: the control group, classical suspension group and modified suspension group (n=30/group). In the classical suspension group, a strip of medical adhesive tape was attached to the tail, with horizontal filament tape wrapping. A piece of gauze was wrapped around the tail at the outermost layer and the tail was suspended for hindlimb unloading. In the modified suspension group, a layer of plastic net was added between the horizontal filament tape and the gauze to reduce the squeeze on the tail as a buffer zone and ensure proper circulation of the tail. After 4 weeks of suspension, damage to the tail and sheath detachment were observed. Meanwhile the body weight and right soleus wet weight of rats were measured. RESULTS: The ratio of right soleus wet weight to body weight was decreased significantly in both the classical suspension group and the modified suspension group compared with the control group, while there was no difference in body weight among the three different groups. Importantly, the incidence of tail ischemia and necrosis (13.3% vs 40.0% in the classical suspension group) and the incidence of sheath detachment from tail (3.3% vs 26.7% in the classical suspension group) were significantly lower whereas the success rates of model (33.3% vs 83.3% in classical suspension group) was significantly higher in the modified suspension group. CONCLUSION The modified protocol decreases the incidence of tail necrosis and sheath detachment in the rat tail suspension and increases the success rate of the hindlimb unloading rat model, with improved simplicity and practicability.
Animals ; Hindlimb Suspension ; Male ; Muscle, Skeletal ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Weightlessness Simulation ; methods

The present study aimed to observe the changes of contractile function and responsiveness to isoproterenol (ISO) in tail-suspended rat cardiomyocytes under simulated weightlessness condition. Tail-suspended rat model was used to simulate weightlessness on the ground. Twenty-four male Sprague-Dawley rats were randomly divided into the control and tail-suspended groups. After 4 weeks of suspension, the rats were injected with heparin (100 IU/100 g body weight, i.p.) and anesthetized with pentobarbital sodium (40 mg/kg body weight). The hearts were removed and the aortas were cannulated rapidly. The cannulated hearts were mounted on a Langendorff perfusion apparatus and perfused with constant flow. The perfusion pressure was monitored. The hearts were digested by 0.08% collagenase I at 37 degrees C. The ventricular tissues were chopped and the single myocytes were dispersed gently by a wide-tipped pipette. The contractile function was measured in the Edge Detector system within 6 h after isolation. The length and width of cardiomyocytes were measured without electric stimulation. Contractile curves of the single cardiomyocytes were recorded at stimulation frequency of 1.0, 2.0 and 4.0 Hz. To observe the responsiveness of cardiomyocytes to ISO, 1, 5 and 10 nmol/L ISO in Kreb's solution was perfused at a stimulation frequency of 2.0 Hz. The length and width of the left and right ventricular cardiomyocytes in tail-suspended group had little difference from that in the control group. The unloaded shortening amplitude increased as stimulation frequency elevated in both the control and tail-suspended groups. It was increased by (8.50±1.26)%, (9.00±1.38)%, (9.23±1.83)% in the left ventricular cardiomyocytes, and (9.80±2.48)%, (10.03±2.48)%, (10.28±2.27)% in the right ventricular cardiomyocytes in the control group at stimulation frequency of 1.0, 2.0 and 4.0 Hz. Compared with that in the control group, the unloaded shortening amplitude decreased by 12.2% and 10.9% in the left ventricular cardiomycytes (P<0.05), and 16.5% and 16.3% in the right ventricular cardiomyocytes (P<0.05) at stimulation frequency of 1.0 and 2.0 Hz in tail-suspended group. There was no significant difference in unloaded shortening amplitude at stimulation frequency of 4.0 Hz between the control and tail-suspended groups. Time to peak shortening (TPS) in tail-suspended group significantly reduced in both the left and right ventricular cardiomyocytes (P<0.05). Time from peak to 75% relaxation (TR(75)) in tail-suspended group significantly prolonged in both the left and right ventricular cardiomyocytes (P<0.05). No significant differences in shortening and relaxation rate (±dL/dt(max)) were observed between the control and tail-suspended groups. The unloaded shortening amplitude increased by (10.63±0.83)%, (35.06±5.22)% and (71.64±6.83)% in the control cardiomyocytes, but increased by (5.75±0.76)%, (23.97±4.50)% and (26.38±8.13)% in tail-suspended group during perfusion with 1, 5 and 10 nmol/L ISO (P<0.05, P<0.01). The unloaded shortening amplitude increased by (3.04±0.27)%, (9.81±2.66)% and (20.20±3.47)% in the control cardiomyocytes, but increased by (1.42±0.53)%, (3.83±1.71)% and (5.49±4.08)% in tail-suspended group during perfusion with 10, 50 and 100 nmol/L forskolin (P<0.05). The results obtained suggest that the unloaded shortening amplitude and responsiveness to ISO decrease in rat cardiomyocytes after 4-week tail-suspension.
Animals ; Electric Stimulation ; Hindlimb Suspension ; Isoproterenol ; pharmacology ; Male ; Myocytes, Cardiac ; drug effects ; Rats ; Rats, Sprague-Dawley ; Weightlessness Simulation

The aim of the present study was to investigate the expressions of calpain and calpastatin in the myocardium of simulated weightlessness rats, and to elucidate the underlying mechanism of cardiac troponin I (cTnI) degradations. Tail-suspended (SUS) rats were used as a simulated weightlessness model on the ground. The myocardium of rats was homogenized, and the expressions of calpain-1, calpain-2, calpastatin and cTnI were analyzed by Western blotting technique. Calpastatin expression was significantly decreased in 2- and 4-week SUS groups compared with that in the synchronous controls (P<0.05). Calpain-2 expression was slightly decreased, whereas calpain-1 expression was unaltered in SUS groups. However, calpain-1/calpastatin and calpain-2/calpastatin ratios were increased after tail-suspension, being significantly higher in 2- and 4-week SUS groups than those in the synchronous controls (P<0.05, P<0.01). Cardiac TnI degradation was significantly increased after tail-suspension (P<0.01), but cTnI degradation in both SUS and control groups was significantly inhibited by a non-specific inhibitor of calpain, PD150606 (P<0.01). These results suggest that an increase in calpain activity may enhance cTnI degradation in the myocardium of tail-suspended rats.
Animals ; Calcium-Binding Proteins ; metabolism ; Calpain ; metabolism ; Hindlimb Suspension ; Myocardium ; metabolism ; Proteolysis ; Rats ; Troponin I ; metabolism ; Weightlessness Simulation

Microgravity is known to produce a number of neurological disturbances during space flight; however, the underlying mechanism of these disturbances is yet to be elucidated. There have been some reports about the increased oxidative stress under microgravity or simulated microgravity. In the present study, we investigated the process of oxidative stress induced by simulated microgravity in different areas of rat brain, which may shed light on the mechanism of neurological disturbances and further neuroprotective research in spaceflight. After adaption for 7 d, 40 healthy male Sprague-Dawley rats were matched for body weight and randomly assigned to control groups (7, 14, 21 and 28 d) and tail-suspended simulated microgravity groups (7, 14, 21 and 28 d). The tail-suspended groups were treated with 30 angels of tail suspension and the control groups were treated similarly to the tail-suspended groups but without tail suspension. After the required times, different structures of rat brain, including cerebellum, cerebral cortex and hippocampus, were harvested and frozen for the further determination. Griess assay, thiobarbituric acid reactive substance (TBARS) assay, competitive ELISA and ferric reducing ability of plasma (FRAP) assay were used for the observation of the changes of reactive nitrogen species (RNS), malondialdehyde (MDA), nitrotyrosine (NT) and total antioxidant capacity (TAC), respectively. As shown in the results, there were different changes in various brain regions after tail suspension compared with control groups. (1) In cerebellum, NT increased after 7 d tail suspension, decreased after 14 d and increased again after 28 d; MDA increased after 14 d; RNS increased and TAC decreased after tail suspension for 21 d; (2) Increase of NT after14 d tail suspension, increase of MDA and decrease of TAC after 21 d were found in cerebral cortex; (3) In hippocampus, RNS increased after tail suspension for 7 d, decreased after 14 d and increased again after 28 d; MDA increased after 21 d; NT increased after 28 d; TAC increased after 7 d and recovered after 21 d. These results suggest that simulated microgravity induced by tail suspension increases the level of oxidative stress in rat brain; however, there are different features in different areas of rat brain. During the response to simulated microgravity, rat brain tissues present a similar process from adaptive response to irreversible oxidative damage.
Animals ; Antioxidants ; metabolism ; Brain ; physiopathology ; Hindlimb Suspension ; Male ; Malondialdehyde ; metabolism ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Weightlessness Simulation

The aim of the present study was to investigate the effect of a short-term (3-day) simulated microgravity with and without daily dorsoventral gravitation (-G(x)) for 1 h on myogenic tone and vasoconstrictor responsiveness of the middle cerebral artery and mesenteric third-order small artery in rats. The tail-suspension (SUS) model was used to simulate cardiovascular deconditioning due to microgravity. Daily restoring to normal standing (STD) posture for 1 h was adopted to provide -G(x) as the countermeasure. Segments of middle cerebral artery and mesenteric third-order small artery were isolated and cannulated. Vascular diameters in response to increased intraluminal pressure (from 20 mmHg to 120 mmHg, by 20 mmHg steps) of isolated arteries under no-flow conditions were recorded by a Pressure Myograph System in both physiologic salt solution (PSS) (active diameter, Da) and calcium-free PSS (passive diameter, Dp). The myogenic tone was calculated by (Dp-Da)/Dpx100%. Vasoconstrictor responsiveness of the isolated middle cerebral artery to serotonin and that of small mesenteric artery to phenylephrine were assessed in the PSS under an intraluminal pressure of 40 mmHg. The results showed that SUS induced an enhancement of the myogenic tone and vasoconstrictor responsiveness in the isolated middle cerebral artery but a depression of those in the small mesenteric artery. Daily STD for 1 h prevented the depression of myogenic tone and vasoconstrictor responsiveness in the small mesenteric artery, but did not prevent the functional enhancement in the middle cerebral artery. These data suggest that a short-term simulated microgravity may result in different alterations in the function of the cerebral artery and the resistance vessel in the hind-body. Moreover, only the decrease of function in these resistance vessels, not in the cerebral arteries, can be prevented by such a countermeasure of daily STD for 1 h.
Animals ; Cerebral Arteries ; pathology ; Hindlimb Suspension ; Mesenteric Arteries ; pathology ; Pressure ; Rats ; Serotonin ; pharmacology ; Vascular Resistance ; Vasoconstriction ; Weightlessness Simulation

OBJECTIVETo study the effects on the male reproduction in adult male rats and its mechanisms through simulated weightlessness using tail-suspension, in order to do a basic works of exploring the effects on human being's reproduction in outer space.

METHODSForty Spraque-Dawley adult male rats were randomly divided into four groups, two experimental groups and two control groups. Rats in the two experimental groups were tail-suspended for 14 d and 28 d respectively, then we examined the weight and morphology of testis, the quality and amount of sperm, also tested the serum hormone by radioimmunoassay and analyzed apoptosis rate of testicular cells by TUNEL in the experimental rats and control rats.

RESULTSAfter tail-suspension, the weight of testis, the sperm count and sperm motility significantly decreased (P <0.05), while the apoptosis rate of testicular cells and the amount of abnormal sperm markedly increased (P <0.05). The content of testosterone significantly decreased (P <0.05), but the contents of FSH and LH mildly increased (P > 0.05). These changes were not significant between two experimental groups (P > 0.05). In addition, the seminiferous tubules became atrophy with the reduction of the layers of seminiferous epithelium, and sperm amount in lumens of seminiferous tubules decreased in experimental groups. The above were more remarkable in the 28 d experimental group.

CONCLUSIONSimulating weightlessness has a harmful effect on reproduction of adult male rats. These may be caused by inducing apoptosis. The blocking apoptosis of testicular cells may be useful in improving the harmful effect.

Animals ; Apoptosis ; Hindlimb Suspension ; Male ; Organ Size ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reproduction ; physiology ; Spermatozoa ; physiology ; Testis ; pathology ; Weightlessness Simulation