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1.
The capacitation process of sperm cells involves complex changes in the composition and orientation of molecules at the surface of the sperm cell. Here we focus on the lipid architecture in the sperm plasma membrane and demonstrate that the sperm plasma membrane is not static but is an extremely dynamic structure. Advanced fluoroscopic techniques enabled continuous monitoring of lipid organization in living cells and extremely rapid lipid movements were observed. The orientation of lipids in the sperm plasma membrane changed under capacitative treatments, was found to be sensitive for temperature and also changed upon binding of sperm cells to the zona pellucida. The changes in membrane properties coincided with an activation of protein kinases resulting in tyrosine phosphorylation of specific plasma membrane proteins. The detected membrane changes relate to intrinsic membrane properties such as fluidity, permeability, adhesiveness and fusibility. We think that these results may provide a physiological basis for new assays, able to discriminate between functional and non-physiological sperm cells.  相似文献   

2.
Summary

The capacitation process of sperm cells involves complex changes in the composition and orientation of molecules at the surface of the sperm cell. Here we focus on the lipid architecture in the sperm plasma membrane and demonstrate that the sperm plasma membrane is not static but is an extremely dynamic structure. Advanced fluoroscopic techniques enabled continuous monitoring of lipid organization in living cells and extremely rapid lipid movements were observed. The orientation of lipids in the sperm plasma membrane changed under capacitative treatments, was found to be sensitive for temperature and also changed upon binding of sperm cells to the zona pellucida. The changes in membrane properties coincided with an activation of protein kinases resulting in tyrosine phosphorylation of specific plasma membrane proteins. The detected membrane changes relate to intrinsic membrane properties such as fluidity, permeability, adhesiveness and fusibility. We think that these results may provide a physiological basis for new assays, able to discriminate between functional and non‐physiological sperm cells.  相似文献   

3.
Based on an experimental model of osteoarthritis in 42 full-grown rabbits the histological changes were studied during the development of osteoarthritis after operative induction of instability of the knee joint. The changes were followed from 2 week to 1½ years after the induction.The first changes were observed at 1 week stage as a proliferation of the lining cells. During the time of observation additional changes were seen such as hypertrophy of villi, infiltrations with plasma cells and lymphocytes, increased vascularity and interstitial edema and fibrosis. Edema was only seen during the first months, later increasing fibrosis was predominant.The first cartilage change was reduction of the staining ability, expressing depletion of GAG, this was seen already at the one week stage whereas morphological changes were present after 4–6 weeks.It was concluded that the synovial membrane in this model shows changes that may contribute to the development of osteoarthritis.  相似文献   

4.
Sperm plasma membrane is a very important structure that functions to protect sperm against extracellular injuries and to respond to physiological challenges. It plays a crucial role during sperm capacitation, in sperm-egg interaction and, finally, in fertilization. Concerning sperm technology, possibly the most important factors causing damage in mammalian spermatozoa membranes are initiated by the osmotic stress generated by dehydration of the cells during freezing and thawing. These changes are rapidly derived to the plasma and organelle membranes that gradually experiment loss of membrane architecture, causing unbalanced production of reactive oxygen species and increased lipid peroxidation. Other procedures such as sperm sorting or liquid storage of sperm also induce harmful changes in the integrity of the membrane. The specific composition of lipids of the sperm membranes may provide clues for understanding the mechanisms behind the differences found in the response to stress in different species. In the present review, we deal with the composition, architecture and organization of the sperm plasma membrane, emphasizing the factors that can affect membrane integrity. The intracellular signalling pathways related with membrane reorganization during capacitation and acrosome reaction are also reviewed.  相似文献   

5.
Reasons for performing study: The timing of lamellar basement membrane (BM) changes occurring during laminitis development is incompletely understood. Objectives: To determine the temporal progression of lamellar BM changes and whether laminin‐332 (Ln‐332) γ2 cleavage products are generated during laminitis development. Methods: Eight clinically normal Standardbred horses were allocated into treatment (n = 5) or sham (n = 3) groups. The treatment group received, via nasogastric intubation, an oligofructose (OF) bolus (10 g/kg bwt) while the sham group was given water. Laminitis induction proceeded for 48 h followed by euthanasia. Lamellar biopsies were obtained prior to dosing and at intervals during the treatment period for analysis (at 12, 18, 24, 30 and 36 h and at 48 h following euthanasia). Results: Changes in lamellar collagen type IV and Ln‐332 were first observed at 12 h post dosing. A unique pattern of reactivity for the Ln‐332 γ2 antibody D4B5 occurred, in which reactivity was observed only in lamellar tissue affected by laminitis. No bioactive Ln‐332 γ2 proteolytic fragments were detected in lamellar samples. Conclusions: Basement membrane changes occurred early during the laminitis process. Direct Ln‐332 γ2 cleavage to release biologically active products did not appear to occur. Thus loss of stability or protein interaction of the BM is probably responsible for the γ2 specific reactivity observed. Potential relevance: Basement membrane changes may a first step in lamellar failure occurring prior to detection with conventional methods. Thus, more sensitive detection methods of BM changes are required to study laminitis development.  相似文献   

6.
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7.
Gentamicin sulfate (3 ml; 50 mg/ml) was administered intra-articularly into 30 normal equine radiocarpal joints after arthrocentesis. Arthrocentesis alone was performed on 10 normal radiocarpal joints. Synovial fluid evaluations and gross and microscopic examinations were performed on synovial fluid and synovial membrane of designated joints at selected daily intervals over a period of 10 days. Synovial fluid from gentamicin-injected joints had greater turbidity, higher RBC and WBC counts, and higher refractive indices than did joints not injected with gentamicin. The largest increases developed on days 1 or 2 after gentamicin injection, with mean total WBC, large mononuclear cell, small mononuclear cell, and polymorphonuclear cell counts of 23,860, 11,853, 857, and 11,150 cells/microliter, respectively. Arthrocentesis alone resulted in smaller increases in these counts. Microscopic changes seen in the synovial membrane of gentamicin-injected joints included edema, leukocytic infiltration, and loss of synovial lining cells. These inflammatory changes resolved within 7 days after gentamicin injection.  相似文献   

8.
Porcine reproductive and respiratory syndrome virus (PRRSV) was isolated from blood samples taken at a pig farm in Hungary from pigs showing clinical signs of the disease. The virus (ABV 32) was identified as belonging to the European genotype by using type-specific monoclonal antibodies. This was confirmed by comparing the sequence of the membrane protein gene (ORF 6) and the nucleocapsid gene (ORF 7) with the American VR2332 and the European LV genotype reference strain, respectively. Analysis of the amino acid sequence of the ORF 6 and ORF 7 of ABV 32 revealed five amino acid changes in both ORFs when compared with LV, of which two changes in ORF 7 were only found in the Spanish isolates. Additionally, the ORF 7 sequence was compared with corresponding sequences of a total of 21 other European strains. Phylogenetic analysis using the PHYLIP package confirmed the close relationship between the Hungarian and the Spanish isolates. Of all the isolates analysed, ABV 32 and LV were the least related.  相似文献   

9.
In osteoarthritis the changes of the synovial membrane may seriously alter the oxygen transfer characteristics from the capillaries of the membrane to the synovial fluid and hence impede or deprive the joint cartilage of its sole source of oxygen. In the present study we have estimated the blood flow (Q), diffusing capacity (DO2) and oxygen consumption (VO2) of the synovial membrane in the chronic non effusive stage of experimental osteoarthritis. In 14 osteoarthritic knee joints we found a statistically significant increase in oxygen consumption, compared to previously reported results from normal joints, whereas the diffusing capacity and the blood flow were unchanged. The implication of this is that the partial pressure difference required to overcome the increased oxygen consumption was increased with a factor 4.3 and that the oxygenation of the joint cartilage was reduced with approx. 30 Torr.  相似文献   

10.
刘艳  岳鑫  陈贵林 《草业学报》2010,19(6):79-86
以一年生乌拉尔甘草为材料,利用透射电镜技术,观察水分胁迫下根皮层细胞和中柱细胞,以及叶片叶肉细胞超微结构的变化,同时结合丙二醛含量测定,分析水分胁迫对甘草根叶细胞超微结构和膜脂过氧化的影响。结果显示,随着干旱胁迫时间的延长,甘草根系、叶片细胞的细胞器结构均发生明显变化,表现为:细胞核变形,染色质凝聚并边缘化;线粒体内腔空化;叶绿体基粒片层结构逐渐模糊,淀粉粒降解;根系中高尔基体扁平囊出芽形成小囊泡等。不同组织以及不同细胞器对干旱胁迫的敏感程度不同,其中根皮层组织的细胞器形态变化早于中柱细胞和叶片。胁迫初期,无论是根系还是叶片,都以细胞核较早发生染色质凝聚,并早于细胞膜脂过氧化损伤。以上特征与细胞程序性死亡特征相似,推测干旱胁迫初期诱导根、叶细胞发生程序性死亡可能是甘草抵御干旱胁迫的机制之一。  相似文献   

11.
低温弱光胁迫下芸豆叶片光抑制与类囊体膜脂构成变化   总被引：1，自引：0，他引：1  
以对低温弱光敏感性有差异的两个红芸豆品种为材料,研究了低温弱光胁迫处理及恢复过程中幼苗叶片的叶绿素含量、荧光参数和类囊体膜脂肪酸组成的变化。结果表明:与对照相比,随胁迫时间延长,叶片中Chl a、Chl b和Chl(a+b)含量降低(P<0.05);Fv/Fm、Fv'/Fm'、qP、ΦPSⅡ和ETR下降(P<0.05),Chl a/b和NPQ上升(P<0.05);类囊体膜脂MGDG、DGDG和SQDG中的亚麻酸(C18:3)含量显著降低(P<0.05),棕榈酸(C16:0)含量显著升高(P<0.05),PG中的棕榈酸(C16:0)和反式十六碳-烯酸[C16:1(3t)]含量降低(P<0.05),而亚麻酸(C18:3)和亚油酸(C18:2)含量升高(P<0.05),在叶片抵御低温弱光胁迫过程中维持一定的膜脂不饱和度的重要作用。随胁迫时间延长,类囊体膜总饱和脂肪酸(SFA)含量升高,多不饱和脂肪酸(PUFA)含量以及膜脂不饱和度(U/S)显著降低,恢复期则相反。在胁迫处理和恢复期,“英大红”和“小红芸豆”的U/S变化差异不明显,PUFA含量变化差异显著(P<0.05),且与Fv/Fm的相关性分别达到86.21%和83.92%,表明低温弱光处理及恢复过程中,光抑制后PSⅡ功能的修复与PUFA含量增加存在一定关系。因此,低温弱光胁迫下“英大红”光抑制程度较“小红芸豆”轻,可能是较高含量的PUFA增加了类囊体膜的不饱和度,维持膜的稳定性,减轻了光抑制。  相似文献   

12.
Degenerative joint disease and inflammation of the synovial membrane were produced in the left stifle of 16 dogs by severing the cranial cruciate ligament. Arthrotomy only was performed on the right stifle. Synovial membrane from these joints was histologically examined at 1, 2, 8, and 13 weeks after surgical operation. Similar tissue was obtained from 4 healthy dogs for comparison. Inflammatory changes in the synovium of the left stifle progressed with time and were prominent at 8 weeks postoperatively; subsynovial fibrosis was greatest at 13 weeks. Inflammation of the synovial membrane and subsynovial tissue was characterized by synovial cell hypertrophy and hyperplasia, plasma cell and lymphocyte infiltration, and increased vascularization of the subsynovial region.  相似文献   

13.
1. Chick embryos of 7, 9, 11, 12, 13, 14, 15, 17 and 19 d of embryonic development were examined to determine the activities of 5-aminolevulinic dehydratase (ALA-D, EC 4.2.1.24) and porphobilinogen deaminase (PBG-D, EC 4.3.1.8). 2. Liver and yolk sac membrane ALA-D specific activities showed a maximum between 12 and 13 d of embryonic development, yolk sac membrane PBG-ase activity a maximum at 9 d and at 7 d in liver. Total activities of ALA-D and PBG-D were not constant during the course of embryonic development but probably related to the changes of intensity of haem synthesis. 3. ALA-D and PBG-ase activities were higher in yolk sac membrane than in liver, showing the importance of the yolk sac membrane as erythropoietic tissue. PBG-D catalysed the rate-limiting reaction of the cytosolic steps in the biosynthetic pathway in both tissues.  相似文献   

14.
Sperm plasma membrane is an essential structure of sperm resistance to freezing. Signs of cryodamage can be visible on the sperm plasma membrane. The aim of our study was to evaluate the appearance of plasma membrane and acrosome in fresh and frozen‐thawed chicken sperm using electron and fluorescence microscopy. Semen was collected from 12 sexually mature roosters of Ross PM3 heavy line, diluted with Kobidil+ extender with 16% of ethylene glycol (KEG; control) or with KEG in combination with one of following non‐permeating cryoprotectants: trehalose (KEG‐TRE) or glycine (KEG‐GLY). Fluorescence staining was used for detection of the membrane integrity, apoptotic changes and viability (Annexin V, Yo‐PRO‐1, PI, respectively). Ultrathin sections (70 nm) from samples were prepared to examine sperm head ultrastructure. Freezing process significantly worsened the status of the sperm plasma membranes. In all frozen groups, only about a quarter of the evaluated sperm were graded as class I quality. In the KEG and KEG‐GLY groups, about half of sperm had severe plasma membrane damages (III class). In sperm with extensively damaged membranes (III class), the acrosome–sperm head junction was mostly disturbed. The use of trehalose was more beneficial (p < 0.05) for sperm plasma membrane than the use of glycine. In contrast, a decrease (p < 0.05) in the apoptotic sperm ratio (Yo‐PRO‐1) was noted in the KEG‐GLY group when compared to other treatments. In conclusion, we identified different plasma membrane and acrosome damages in cryopreserved chicken sperm. The loss of acrosomes can contribute to diminishing of fertilization ability of cryopreserved chicken sperm.  相似文献   

15.
1. Oxygen permeabilities (KO2 ) of the shell and shell membranes of fertile and infertile chicken eggs were measured at 37.5 °C and a relative humidity of 0.60 throughout 14 d incubation, with turning. The KO2 of the shell and membranes of infertile eggs was around 1.0 × 10?7 cm3 O2STP sec?1 cm?2 Torr?1 (1 Torr = 133.322 Pa) throughout incubation. With fertile eggs, from which there was a linear loss of water during incubation, the KO2 of the shell and shell membranes was about 1.0 × 10?7 cm3 O2STP sec?1 cm?2 Torr?1 for the first four days of incubation. Thereafter the majority of shells and membranes had a Ko2 of about 1.0 × 10?6 cm3 O2STP sec?1 cm?2 Torr?1.

2. A diminution of the Na+ and K+ content of the shell membranes of fertile eggs was not associated with changes in the dimensions of the glyco‐protein mantle on the cores of the individual fibres of the membranes. There was, however, a progressive deterioration in the limiting membrane of fertile but not of infertile eggs.

3. It was concluded that changes in the O2 resistance of the integument of fertile eggs were not a product of change in either of the shell membranes but of damage caused to the limiting membrane by the chorioallantois.  相似文献   


16.
Our previous work has shown that an anti-androgen flutamide administered pre- and post-natally induced adverse effects on the epididymal morphology and function of adult boars. The present investigation is aimed to understand the effect of flutamide and its metabolite on changes in sperm plasma membrane integrity and its stability, changes in mitochondrial oxidative capability and frequency of abnormal sperm. In vivo effects of flutamide (50 mg/kg b.w.) on sperm ultrastructure were examined by electron microscopic observations. In vitro effects of 5, 50 and 100 μg/ml hydroxyflutamide, administered for 2 and 24 h, on sperm plasma membrane integrity were measured by LIVE/DEAD Sperm Vitality kit, while those on sperm membrane stability and mitochondrial oxidoreductive activity were investigated using Merocyanine 540 and NADH tests, respectively. The incidence of abnormal spermatozoa increased significantly (p < 0.05) in flutamide-treated boars compared with controls. In an in vitro approach, low dose of hydroxyflutamide in 2-h incubations appeared less effective in altering the sperm plasma membrane integrity and its stability than two higher doses used (p < 0.05). No further decrease in the membrane integrity was found when the effect of anti-androgen lasted for 24 h. On the other hand, a decrease in sperm membrane destabilization and mitochondrial oxidoreductive activity was strengthened after 24 h of hydroxyflutamide administration (p < 0.05). Characterization of sperm parameters with regard to oxidative capability of mitochondria, plasma membrane changes and sperm ultrastructure provides novel data on the boar sperm sensitivity to anti-androgen action. Results indicate high sensitivity of boar spermatozoa to androgen withdrawal.  相似文献   

17.
Autoantibodies and erythrocyte membrane proteins were analyzed in a case of a dog with autoimmune hemolytic anemia (AIHA). In crisis phase, antiglobulin test was positive. The eluate from the erythrocytes of the dog with AIHA gave agglutination against autologus erythrocytes. Immunoglobulin subclasses in the eluate were revealed to be IgG and IgA by the double diffusion test. Comparing the SDS polyacrylamide gel electrophoretic patterns of erythrocyte membrane proteins between the crisis and remission phases, there was a change in the protein on protein 4.1 region. However, there were no changes in blood group typings in two phases.  相似文献   

18.
The SDS-PAGE patterns of the outer membrane protein (OMP) extracts of Pasteurella multocida strain P1059, grown under iron-restricted, iron-replete and in vivo conditions, were examined. The results showed that the iron-regulated outer membrane proteins (IROMPs) with molecular masses of 76 kDa, 84 kDa, and 94 kDa were expressed by bacteria grown in iron-restricted media. They were also expressed by in vivo grown P. multocida. Convalescent-phase sera, obtained from turkeys which had survived pasteurellosis, contained antibodies that reacted intensly with th three IROMPs. This indicated that these proteins were expressed in vivo. Bacteria expressing the IROMPs showed greater binding to Congo Red when compared to cells not expressing IROMPs. Cells expressing the IROMPs or its OMP extracts grown in iron-restricted media also showed greater binding to 59Fe-pasteurella siderophore (multocidin) when compared to bacteria or its extracts not expressing IROMPs. Convalescent-phase sera, which contained antibodies against the IROMPs, blocked this specific 59Fe-multocidin binding to IROMPs. Autoradiography was used to determine which of these IROMPs functioned as a receptor for the iron-multocidin complex. The results suggested that these three IROMPs have specific epitopes for binding to the iron multocidin complex.  相似文献   

19.
The oxygen supply to the joint cartilage depends on the oxygen transport from the capsular arteries to the capillaries, the oxygen diffusion across the synovial membrane and the oxygen transport through the synovia. In osteoarthritis the resistance to transport across all 3 barriers may be increased because of the joint effusion, the elevated intraarticular pressure and the inflammatory changes of the synovial membrane.In the present study we describe a method to determine 2 important parameters affecting the oxygen transport through the synovial membrane: the oxygen consumption and diffusion capacity of the membrane. The principle of the method is to perfuse the joint cavity of the knee by saline saturated with air and to record the relationship between the oxygen partial pressure in the outflowing perfusate and the perfusion rate.The values found for the diffusion capacity and oxygen consumption were 0.039 ± 0.013 μl O2/min/Torr and 0.93 ± 0.90 μl O2/min (mean ± s).  相似文献   

20.
Concanavalin-A points of linkage were positively detected on cell membranes of alveolar and peritoneal macrophages by means of Con-A ferritin conjugate. Quantitative conclusions were drawn from these findings with regard to the number of mannose and glucose residues per 1 micron 2 of membrane area. With different incubation periods, 15, 25, and 45 minutes, various distribution patterns of ferritin molecules were recorded. They were diffusely distributed in cytoplasma as well as on the outer nuclear membrane. Ferritin particles were identified also on vacuolar membranes and in direct contact with lysosomes.  相似文献   

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