Cholesterol is an essential component of the mammalian plasma membrane because it promotes membrane stability without comprising membrane fluidity. Given this important cellular role, cholesterol levels are tightly co...Cholesterol is an essential component of the mammalian plasma membrane because it promotes membrane stability without comprising membrane fluidity. Given this important cellular role, cholesterol levels are tightly controlled at multiple levels. It has been clearly shown that cholesterol redistribution and depletion from the sperm membrane is a key part of the spermatozoon＇s preparation for fertilization. Some factors that regulate these events are described （e.g., bicarbonate, calcium） but the mechanisms underlying cholesterol export are poorly understood. How does a hydrophobic cholesterol molecule inserted in the sperm plasma membrane enter the energetically unfavorable aqueous surroundings？ This review will provide an overview of knowledge in this area and highlight our gaps in understanding. The overall aim is to better understand cholesterol redistribution in the sperm plasma membrane, its relation to the possible activation of a cholesterol transporter and the role of cholesterol acceptors. Armed with such knowledlze, sl）erm handlin~ techniques can be adapted to better prepare spermatozoa for in vitro and in vivo fertilization.展开更多
Amyloid-beta peptide is the main component of amyloid plaques, which are found in Alzhei- mer＇s disease. The generation and deposition of amyloid-beta is one of the crucial factors for the onset and progression of Al...Amyloid-beta peptide is the main component of amyloid plaques, which are found in Alzhei- mer＇s disease. The generation and deposition of amyloid-beta is one of the crucial factors for the onset and progression of Alzheimer＇s disease. Lipid rafts are glycolipid-rich liquid domains of the plasma membrane, where certain types of protein tend to aggregate and intercalate. Lipid rafts are involved in the generation of amyloid-beta oligomers and the formation of amyloid-beta peptides. In this paper, we review the mechanism by which lipid rafts disturb the aberrant deg- radative autophagic-lysosomal pathway of amyloid-beta, which plays an important role in the pathological process of Alzheimer＇s disease. Moreover, we describe this mechanism from the view of the Two-system Theory of fasciology and thus, suggest that lipid rafts may be a new target of Alzheimer＇s disease treatment.展开更多
Aim： In the present study, a variety of high resolution microscopy techniques were used to visualize the organization and motion of lipids and proteins in the sperm＇s plasma membrane. We have addressed questions suc...Aim： In the present study, a variety of high resolution microscopy techniques were used to visualize the organization and motion of lipids and proteins in the sperm＇s plasma membrane. We have addressed questions such as the presence of diffusion barriers, confinement of molecules to specific surface domains, polarized diffusion and the role of cholesterol in regulating lipid rafts and signal transduction during capacitation. Methods： Atomic force microscopy identified a novel region （EqSS） within the equatorial segment of bovine, porcine and ovine spermatozoa that was enriched in constitutively phosphorylated proteins. The EqSS was assembled during epididymal maturation. Fluorescence imaging techniques were then used to follow molecular diffusion on the sperm head. Results： Single lipid molecules were freely exchangeable throughout the plasma membrane and showed no evidence for confinement within domains. Large lipid aggregates, however, did not cross over the boundary between the post-acrosome and equatorial segment suggesting the presence of a molecular filter between these two domains. Conclusion： A small reduction in membrane cholesterol enlarges or increases lipid rafts concomitant with phosphorylation of intracellular proteins. Excessive removal of cholesterol, however, disorganizes rafts with a cessation of phosphorylation. These techniques are forcing a revision of long-held views on how lipids and proteins in sperm membranes are assembled into larger complexes that mediate recognition and fusion with the egg. （Asian JAndrol 2007 July; 9： 438-444）展开更多
Lipid rafts are sterol and sphingolipid rich membrane domains that possibly may play roles in multiple cellular processes. These domains are still the matter of debate and it is still unknown by which mechanism if any...Lipid rafts are sterol and sphingolipid rich membrane domains that possibly may play roles in multiple cellular processes. These domains are still the matter of debate and it is still unknown by which mechanism if any and organisms promote their formation. This study centers on the ease of in vitro formation of lipid rafts-like structures as it relates to the relative availability of sphingolipids, phospholipids, cholesterol, and membrane proteins. Following a 12 h incubation period, isolation and extraction of the lipid rafts-like assemblies, the composition of the structures was evaluated using HPLC. Cholesterol and sphingomyelin were detected at 206 nm and phosphatidylcholine was detected at 254 nm. Identification of lactose permease, a typical membrane protein, was done using FTIR. The thermal stability of the produced structures was also determined. Results show that the addition of cholesterol significantly increased both the amount of insoluble lipid rafts-like structures and their stability, and that the availability of a minimum amount of sphingolipid was necessary to produce larger amounts of more stable structures. However, the addition of phospholipids hindered the formation of lipid rafts-like assemblies and those formed were generally less stable.展开更多
For a long time, lipid rafts have been thought to participate in regulating neurotransmitter release. However, the existence of lipid rafts on synaptic vesicles （SVs） and the mechanism by which exocytosisrelative pr...For a long time, lipid rafts have been thought to participate in regulating neurotransmitter release. However, the existence of lipid rafts on synaptic vesicles （SVs） and the mechanism by which exocytosisrelative proteins distribute on this structure have not been fully investigated. There is also much controversial data concerning rafts on SVs and synaptic vesicle proteins which makes the results difficult to interpret. This study systematically analyzed the existence and properties of lipid rafts on purified SVs by sucrose density gradient centrifugation, cholesterol depletion, and temperature variation. The data reveals that typical lipid rafts on SVs are both cholesterol dependent and temperature sensitive. Previous confusing results may have been caused by improper treatment or side effects of particular reagent. We also screened the lateral distribution of major exocytosis-related SV proteins and found that only the synaptobrevin （syb） and synaptotagmin （syt） produce detectable association with lipid rafts in 1% Triton X-100.展开更多
文摘Cholesterol is an essential component of the mammalian plasma membrane because it promotes membrane stability without comprising membrane fluidity. Given this important cellular role, cholesterol levels are tightly controlled at multiple levels. It has been clearly shown that cholesterol redistribution and depletion from the sperm membrane is a key part of the spermatozoon＇s preparation for fertilization. Some factors that regulate these events are described （e.g., bicarbonate, calcium） but the mechanisms underlying cholesterol export are poorly understood. How does a hydrophobic cholesterol molecule inserted in the sperm plasma membrane enter the energetically unfavorable aqueous surroundings？ This review will provide an overview of knowledge in this area and highlight our gaps in understanding. The overall aim is to better understand cholesterol redistribution in the sperm plasma membrane, its relation to the possible activation of a cholesterol transporter and the role of cholesterol acceptors. Armed with such knowledlze, sl）erm handlin~ techniques can be adapted to better prepare spermatozoa for in vitro and in vivo fertilization.
基金supported by a grant from Projects of High-tech Industrialization of Guangdong Province of China,No.2011B010500004a grant from National Financial Major Project of China
文摘Amyloid-beta peptide is the main component of amyloid plaques, which are found in Alzhei- mer＇s disease. The generation and deposition of amyloid-beta is one of the crucial factors for the onset and progression of Alzheimer＇s disease. Lipid rafts are glycolipid-rich liquid domains of the plasma membrane, where certain types of protein tend to aggregate and intercalate. Lipid rafts are involved in the generation of amyloid-beta oligomers and the formation of amyloid-beta peptides. In this paper, we review the mechanism by which lipid rafts disturb the aberrant deg- radative autophagic-lysosomal pathway of amyloid-beta, which plays an important role in the pathological process of Alzheimer＇s disease. Moreover, we describe this mechanism from the view of the Two-system Theory of fasciology and thus, suggest that lipid rafts may be a new target of Alzheimer＇s disease treatment.
文摘Aim： In the present study, a variety of high resolution microscopy techniques were used to visualize the organization and motion of lipids and proteins in the sperm＇s plasma membrane. We have addressed questions such as the presence of diffusion barriers, confinement of molecules to specific surface domains, polarized diffusion and the role of cholesterol in regulating lipid rafts and signal transduction during capacitation. Methods： Atomic force microscopy identified a novel region （EqSS） within the equatorial segment of bovine, porcine and ovine spermatozoa that was enriched in constitutively phosphorylated proteins. The EqSS was assembled during epididymal maturation. Fluorescence imaging techniques were then used to follow molecular diffusion on the sperm head. Results： Single lipid molecules were freely exchangeable throughout the plasma membrane and showed no evidence for confinement within domains. Large lipid aggregates, however, did not cross over the boundary between the post-acrosome and equatorial segment suggesting the presence of a molecular filter between these two domains. Conclusion： A small reduction in membrane cholesterol enlarges or increases lipid rafts concomitant with phosphorylation of intracellular proteins. Excessive removal of cholesterol, however, disorganizes rafts with a cessation of phosphorylation. These techniques are forcing a revision of long-held views on how lipids and proteins in sperm membranes are assembled into larger complexes that mediate recognition and fusion with the egg. （Asian JAndrol 2007 July; 9： 438-444）
文摘Lipid rafts are sterol and sphingolipid rich membrane domains that possibly may play roles in multiple cellular processes. These domains are still the matter of debate and it is still unknown by which mechanism if any and organisms promote their formation. This study centers on the ease of in vitro formation of lipid rafts-like structures as it relates to the relative availability of sphingolipids, phospholipids, cholesterol, and membrane proteins. Following a 12 h incubation period, isolation and extraction of the lipid rafts-like assemblies, the composition of the structures was evaluated using HPLC. Cholesterol and sphingomyelin were detected at 206 nm and phosphatidylcholine was detected at 254 nm. Identification of lactose permease, a typical membrane protein, was done using FTIR. The thermal stability of the produced structures was also determined. Results show that the addition of cholesterol significantly increased both the amount of insoluble lipid rafts-like structures and their stability, and that the availability of a minimum amount of sphingolipid was necessary to produce larger amounts of more stable structures. However, the addition of phospholipids hindered the formation of lipid rafts-like assemblies and those formed were generally less stable.
基金Supported by the National Key Basic Research and Development (973) Program of China (No. 2004CB720005)
文摘For a long time, lipid rafts have been thought to participate in regulating neurotransmitter release. However, the existence of lipid rafts on synaptic vesicles （SVs） and the mechanism by which exocytosisrelative proteins distribute on this structure have not been fully investigated. There is also much controversial data concerning rafts on SVs and synaptic vesicle proteins which makes the results difficult to interpret. This study systematically analyzed the existence and properties of lipid rafts on purified SVs by sucrose density gradient centrifugation, cholesterol depletion, and temperature variation. The data reveals that typical lipid rafts on SVs are both cholesterol dependent and temperature sensitive. Previous confusing results may have been caused by improper treatment or side effects of particular reagent. We also screened the lateral distribution of major exocytosis-related SV proteins and found that only the synaptobrevin （syb） and synaptotagmin （syt） produce detectable association with lipid rafts in 1% Triton X-100.
