Unloading of the soleus in tail-suspended rats increases the number of intrafusal fibers in muscle spindles

Transmission electron microscopy was used to study the ultrastructure of muscle spindles (encapsulated stretch receptors) in m. soleus of adult Wistar rats after repeated hindlimb unloading. It was shown that the unloaded soleus contained not only spindles with a typical number of intrafusal fibers (four) but also spindles with five or six fibers. The increase in the number of intrafusal fibers in muscle spindles of the unloaded animals is likely to be caused by the proliferation of their satellite cells (myoblasts).     

Differential effects of low-magnitude high-frequency vibration on reloading hind-limb soleus and gastrocnemius medialis muscles in 28-day tail-suspended rats

Objectives:Low-magnitude high-frequency vibration (LMHFV) was reported beneficial to muscle contractile functions in clinical and preclinical studies. This study aims to investigate the effects of LMHFV on myofibers, myogenic cells and functional properties of disused soleus (Sol) and gastrocnemius medialis (GM) during reloading.Methods:Sprague Dawley rats were hind-limb unloaded for 28 days and assigned to reloading control (Ctrl) or LMHFV group (Vib). Sol and GM of both groups were harvested for fiber typing, proliferating myogenic cell counting and in vitro functional assessment.Results:Myogenic cells proliferation was promoted by LMHFV in both Sol and GM (p<0.001 and p<0.05 respectively). Force generating capacity was not much affected (Vib=Ctrl, p>0.05) but fast-fiber favorable changes in fiber type switching (more type IIA but lower type I in Vib; p<0.05 and 0.01 respectively) and fiber hypertrophy (type I, Vib<Ctrl; p<0.01) were observed mainly in GM.Conclusion:LMHFV was not detrimental to reloading muscles but the outcomes were muscle dependent. The unique fiber type composition and anatomical differences between Sol and GM might render the differential muscle responses to LMHFV. Further investigations on myofibers type specific responses to different LMHFV regimes and myogenic cell interaction with associated myofiber were proposed.     

Cerebral arteriolar structure and function in pinealectomized rats

We examined cerebral arteriolar structure and autoregulation of cerebral blood flow (CBF) in control (n = 8), sham-operated (n = 8), pinealectomized (n = 10), and pinealectomized plus melatonin-treated (0.51 +/- 0.01 mg x kg(-1) x day(-1) in drinking water, n = 9) young Wistar rats. The lower limit of CBF autoregulation (LLCBF) was determined by measurement of CBF (in arbitrary units, laser Doppler) during stepwise hypotensive hemorrhage; the arteriolar internal diameter (ID; in microm, cranial window) was also measured. Measurements of ID were repeated during a second stepwise hypotension after smooth muscle cell deactivation (67 mmol/l EDTA). The cross-sectional area (CSA) was measured by histometry. CSA and EDTA-induced vasodilatation decreased after pinealectomy (517 +/- 21 vs. 819 +/- 40 microm(2) in sham and 829 +/- 55 microm(2) in control, P < 0.05, and 81 +/- 4 vs. 102 +/- 5 microm in sham and 104 +/- 4 microm in control, P < 0.05, respectively) and were restored by melatonin (924 +/- 39 microm(2) and 102 +/- 5 microm, respectively). These results suggest that melatonin deprival makes the arteriolar wall thinner and stiffer. However, these changes had little effect on LLCBF. In conclusion, pinealectomy of young rats induces atrophy and decreases distensibility of the cerebral arteriolar wall; these effects are prevented by melatonin. They do not modify LLCBF.     

Changes in blood biochemical parameters in tail-suspended rats

There have been many studies on the effects of gravity on animals and humans. In particular, weightlessness and the time course of physiological shifts in space acclimation are the most significant problems in a space environment. In a laboratory setting, tail suspension in rats has been utilized for simulation of weightlessness. We therefore utilized this technique in order to study the influence of microgravity on blood constituents in rats. It was clarified that some blood biochemical parameters were effected for the duration of suspension exposure, and they are reported here.     

Changes of angiotensinogen expression in arteries of tail-suspended rats

Previous findings from our laboratory have demonstrated that simulated microgravity may result in atrophic changes with depressed vasoconstrictor responsiveness in hindquarter vessels, and hypertrophic changes with enhanced vasoconstrictor responsiveness in cerebral arteries of rats. However, the mechanisms of this differential adaptation are still not well understood. Local renin-angiotensin system (L-RAS) has been found to be actively involved in the remodeling of arteries. We hypothesized that L-RAS may function as a local regulatory mechanism in the microgravity-induced differential changes of arterial vessles. Angiotensinogen (AGT) is the only and indispensable substrate of local renin-angiotensin system (L-RAS). In the present work, the expression changes of AGT mRNA and protein level as well as its time course characteristics were examined.     

钙蛋白酶介导模拟失重大鼠心肌肌钙蛋白抑制亚基的降解

本文旨在观察尾部悬吊模拟失重大鼠心肌钙蛋白酶(calpain)与钙蛋白酶抑素(calpastatin)表达的变化,以探讨心肌肌钙蛋白抑制亚基(cardiac troponin I,cTnI)降解的可能机制。采用尾部悬吊模拟失重大鼠模型,Western blotting技术观测心肌calpain-1、calpain-2与calpastatin的表达;PD150606抑制calpain活性,分析cTnI降解程度的变化。结果显示:与同步对照组相比,悬吊2周与4周组大鼠心肌calpastatin表达呈显著性降低(P0.05),calpain-1表达未改变,calpain-2表达略有降低;但是,心肌calpain-1/calpastatin及calpain-2/calpastatin的比值在悬吊2周与4周组明显增高(P0.05,P0.01)。悬吊4周组cTnI降解显著高于对照组(P0.01);然而,用calpain非特异性抑制剂PD150606处理后,对照组及悬吊组cTnI的降解均被显著抑制(P0.01)。这些结果提示模拟失重大鼠心肌calpain活性增高可能增加cTnI的降解。     

Proteomic analysis of pulmonary tissue in tail-suspended rats under simulated weightlessness

Weightlessness affects lung function and even causes certain damages to pulmonary tissue. This study used rat tail-suspension model to simulate the physiological effects of weightlessness and investigate the alterations of lung proteome, to reveal the mechanism of lung injury under weightlessness condition. Twenty male Sprague-Dawley rats were randomly divided into two groups: tail-suspended and control. Protein samples from pulmonary tissue of tail-suspended and control groups were separated by two-dimensional (2D) gel electrophoresis and analyzed with ImageMaster 2D elite software. Differentially expressed proteins were identified by high definition mass spectrometry (HDMS) in combination with database searching. Seventeen differentially expressed proteins were identified, among which 13 proteins were upregulated, and four proteins downregulated. The functions of these identified proteins can be classified into six classes related to: metabolism, oxidative stress, cellular functions, cytoskeletal proteins, signal tranduction, and protein degradation. They are mainly related to cellular energy metabolism, stress and inflammatory response, cell injury and repair, intracellular signal transduction, and other cellular functions, playing important roles in weightlessness-induced lung injury.     

Peroxynitrite induces arteriolar smooth muscle cells membrane hyperpolarization with arteriolar hyporeactivity in rats

Peroxynitrite (ONOO-) has been recently known to act as a potent cytotoxin during pathogenesis of various diseases. This study aimed to investigate the possible effect of ONOO- on the cremaster muscle arteriolar reactivity in response to noradrenaline and subsequently determined whether membrane hyperpolarization and potassium channel activation were involved in ONOO(-)-induced alteration of arteriolar reactivity. The results demonstrated that 1) ONOO- could decrease arteriolar reactivity in a time- and concentration-dependent manner with no significant alteration of arteriolar diameter; 2) Superfusion with 20 microM ONOO- over 40 minutes showed slight but not significant influence on the resting potential (Em) of arteriolar smooth muscle cells (ASMCs). However, ASMCs subjected to 50 or 100 microM ONOO- administration were significantly hyperpolarized. As control, treatment with 50 microM decomposed ONOO- or Kreb's solution had little effect on the Em of ASMCs; 3) ONOO(-)-induced arteriolar hyporeactivity could be greatly reversed by co-administration of KCl and partially by TEA. The above results indicated that membrane hyperpolarization and potassium channel activation were preferentially responsible for the reduction of cremaster muscle arteriolar reactivity after exposure to ONOO-.     

The effect of Lodronat(R) on development of osteoporosis in tail-suspended rats

It has been recently revealed that deficient loading of the musculoskeletal apparatus gives rise to osteoporosis. It is also a common knowledge, that diphosphonate preparations are used to advantage in clinical practice to prevent and treat osteoporosis of varying etiology. In connection with the above an attempt was made to evaluate potentials of dichlormethylene diphosphonic acid (Lodronat(R), Boehringer Mannheim) as a countermeasure against osteoporosis in tail-suspended rats (micro-g simulation).     

Vasoconstrictor responsiveness of hind body vascular beds is diminished in tail-suspended rats

It is well known that the mechanisms of occurrence of orthostatic intolerance induced by exposure to microgravity deal with multiple factors including alterations of arteries. In the previous works, the diminished contractile responsiveness of abdominal aorta and hind body medium-sized conduit arteries, mesenteric artery and femoral artery, were observed in tail-suspended rats, and the data showed that the femoral artery have subjected to the greatest changes. These results suggested that the vasoreactivity of resistance vessels might be affected by the real or simulated microgravity. Since the arterioles are the main site of peripheral resistance and of its regulation. Therefore, changes in responsiveness of arteriolar network, especially in the lower/hind body region, would be of primary importance in the genesis of postflight orthostatic intolerance. The aim of the present work was to examine whether simulated weightlessness may lead to an impairment in vasoconstrictor responsiveness in hind body vascular beds.     

Daily short-term head-up tilt can prevent structural changes in arteries of tail-suspended rats

The aim of the present study was to investigate the effectiveness of daily 4-h head up tilt (HUT) in preventing the structural changes in arteries from rats induced by tail-suspension (TS). TS rat model was used to simulate the physiological effects of microgravity. Daily 4-h HUT was used to simulate the effect of intermittent artificial gravity (IAG). The results showed that TS alone induced an increase in the vessel media layer cross-sectional area (CSA), media wall thickness (T), mean number of smooth muscle cell layers (NCL) in basilar artery, and a decrease in these parameters in anterior tibial artery. Both the hypertrophic and atrophic changes in these arteries that might occur due to TS alone can be prevented by a 4h/d HUT treatment.     

Changes in total fiber numbers of disused soleus muscle on rats

In this study, by use of technique that was modified from Morey method, we discussed the histological influence on the soleus muscle of the rats caused by disuse. This study is characterized by the calculating of total numbers of muscle fibers. ST (slow-twitch) and FT (fast-twitch) fibers in total muscular cross-sectional area were classified by their difference in intensity of staining of actomyosin adenosinetriphosphatase (myosin ATPase). During the experiment, average fiber diameter of ST and FT fibers declined when compared to control group (p less than 0.01). A 54% decrease in the total number of ST fibers was observed in the experimental group (p less than 0.01). Conversely, the total number of FT fibers increased to 362% of the control value (p less than 0.01). These results of the changes evoked in ST and FT fibers indicate 34% decrease in total muscular cross-sectional area, and showed that muscular function shifted toward a faster muscle in disused soleus muscle.     

Nuclear translocation of calpain-2 regulates propensity toward apoptosis in cardiomyocytes of tail-suspended rats

The compensatory increase in catecholamine release does not reverse orthostatic intolerance after returning from a long-term spaceflight, but it is unclear whether high dose of catecholamine induces cardiac damage. The tail-suspended rat model was used to simulate the effects of weightlessness on the heart. Apoptotic rates in the left ventricular myocardium did not increase in 4-week of tail-suspended rats compared with the synchronous control. On the contrary, isoproterenol (intraperitoneal injection) and 1-day recovery from the 4-week tail-suspension increased apoptotic rates in the myocardium. Propranolol and PD150606 inhibited cardiomyocyte apoptosis in the recovery group. PD150606 and calpain-2 knockdown also blocked isoproterenol-induced cardiomyocyte apoptosis in tail-suspended rats. The activity and nuclear translocation of calpain-2 increased, but the expression of calpain-1, calpain-2, and calpastatin was unchanged in the myocardium of tail-suspended rats. The Ser-16-phosphorylated phospholamban of the nuclear envelope was higher in tail-suspended rats than in the control rats under isoproterenol stimulation. Isoproterenol treatment also induced a large intranuclear Ca(2+) transient of cardiomyocytes in tail-suspended rats. These results suggest that high-dose isoproterenol phosphorylates phospholamban of the nuclear envelope and increases intranuclear Ca(2+) transient. Larger intranuclear Ca(2+) further activates nuclear calpain-2 and hence induces cardiomyocyte apoptosis.     

Observations on the structure of pullulan

An extracellular α-glucan, pullulan, elaborated by a strain of Pullularia pullulans, contains 0.6% of a maltotetraose subunit, as well as the already known major component, maltotriose. The majority, at least, of the maltotetraose is contained within the polymeric chains and is linked, through its terminal glucose units, by α-1→6— bonds.     

Observations on structure and differentiation in plasmodesmata

Summary The fine structure of plasmodesmata in a number of plant tissues has been examined following fixation in glutaraldehyde. The structure of plasmodesmata is not constant. Variations occur between species, tissues and between different cellular situations in a single tissue. The nature of these variations is described and related to current theories of the formation and function of plasmodesmata. Evidence is presented that the young cell wall after division contains a large number of plasmodesmata, probably functionally and structurally identical, and that the development process involves characteristic modifications both to the distribution of plasmodesmata within the wall, and to the structure of individual plasmodesmata. The probable importance of the endoplasmic reticulum is stressed in relation to the formation and functioning of plasmodesmata.     

ADP-induced pial arteriolar dilation in ovariectomized rats involves gap junctional communication

It was previously shown that, despite the loss of nitric oxide (NO) dependence, ADP-induced pial arteriolar dilation was not attenuated in estrogen-depleted [i.e., ovariectomized (Ovx)] rats. Additional evidence suggested that the NO was replaced by an endothelium-dependent hyperpolarizing factor (EDHF)-like mechanism. To further characterize the nascent EDHF role in Ovx females, the current study was undertaken to test whether, in Ovx rats, ADP-induced pial arteriolar dilation retained its endothelial dependence and whether gap junctions are involved in that response. A closed cranial window and intravital microscopy system was used to monitor pial arteriolar diameter changes in anesthetized rats. The endothelial portion of the ADP-induced dilation was evaluated using light dye endothelial injury (L/D). The study was organized around three experimental approaches. First, the responses of pial arterioles to ADP before and after L/D exposure in intact and Ovx female rats were tested. L/D reduced the ADP response by 50-70% in both groups, thereby indicating that the endothelium dependence of ADP-induced vasodilation is not altered by chronic estrogen depletion. Second, the NO synthase inhibitor N(omega)-nitro-L-arginine (L-NNA) and the prostanoid synthesis inhibitor indomethacin (Indo) were coapplied. In intact females, L-NNA-Indo attenuated the response to ADP by 50%, with no further changes upon the addition of L/D. On the other hand, L-NNA-Indo did not affect ADP reactivity in Ovx rats, but subsequent L/D exposure reduced the ADP response by >50%. The NO-prostanoid-independent, but endothelium-dependent, nature of the response in Ovx females is a hallmark of EDHF participation. Third, gap junctional inhibition strategies were applied. A selective inhibitor of gap junctional function, Gap 27, did not affect ADP reactivity in intact females but reduced the the ADP response by 50% in Ovx females. A similar result was obtained following application of a connexin43 antisense oligonucleotide. These findings suggest that the nascent EDHF dependency of ADP-induced pial arteriolar dilation in Ovx females involves connexin43-related gap junctional communication.     

Observations on the structure of bacilysin.

1. Elementary analysis and other properties of a highly purified preparation of bacilysin indicated that a possible molecular formula for the substance is C(12)H(18)N(2)O(5). The results of electrometric titration were consistent with the hypothesis that the substance was a peptide containing one free alpha-amino group and one free carboxyl group. 2. Hydrolysis of bacilysin with 6n-hydrochloric acid at 105 degrees yielded l-alanine and l-tyrosine, but the ultraviolet spectrum of the substance showed that no tyrosine residue was present in the molecule and a nuclear-magnetic-resonance spectrum indicated that olefinic and aromatic protons were absent. The dinitrophenyl (DNP) derivative of bacilysin yielded DNP-alanine on acid hydrolysis. 3. Bacilysin was hydrolysed by leucine aminopeptidase (EC 3.4.1.1) and by Pronase to give alanine and an uncharacterized amino acid. Its infrared spectrum was consistent with the presence of a peptide grouping in the molecule. 4. The optical rotatory dispersion of bacilysin and its reaction with thiosemicarbazide indicated that the substance contained an aldehyde or ketone group. Its behaviour on catalytic reduction and its reaction with sodium thiosulphate and with certain thiols suggested that an epoxide group was present. 5. A possible type of structure for bacilysin is considered in the light of its known properties.