Biomaterial Database

Isolation and characterization of the plasma membrane of rat cauda epididymal spermatozoa. 1988

P Agrawal, and S F Magargee, and R H Hammerstedt

Biochemistry Program, Pennsylvania State University, University Park 16802.

Cauda epididymal rat spermatozoa were isolated by flushing the excised epididymis and the plasma membrane was detached by a nitrogen cavitation treatment (500 psi, 10 minutes equilibration at 4 C). Membrane vesicles were recovered after sucrose gradient centrifugation. Portions of the sperm surface releasing the plasma membrane were assessed by light microscopy of fluoroscein isothiocyanate-succinylated concanavalin A-treated spermatozoa and by transmission electron microscopy. Plasma membrane was detached from the region overlying the acrosome from most spermatozoa and from the middle-piece overlying the mitochondria from some cells. Thus, the fraction analyzed was derived from at least two portions of the sperm surface. The fractions from the sucrose density gradient were analyzed for gross chemical composition (phospholipid, protein and sterol) and the protein components were detected after electrophoresis under denaturing conditions; the peak fractions (at density approximately 1.13 g/ml) were judged homogeneous. Replicate analyses of such preparations established mass ratios of protein to phospholipid of 0.63, total sterol to phospholipid of 0.18, and demosterol to cholesterol of 0.32. The molecular composition of the phospholipid fraction was determined to be 10% phosphatidylserine (mole percent), 3% phosphatidylinosital, 3% sphingomyelin, 31% phosphatidylethanolamine, 27% phosphatidylcholine, 10% diphosphatidylglycerol and 5% of an unknown component. Fatty acyl analyses of the phospholipid fraction revealed that approximately 70% of the residues consisted of palmitoyl (16:0) and stearoyl (18:0) acyl groups, with the balance distributed among various unsaturated acyl groups (18:1, 22:3, 22:4 and 22:5); about 40% of the recovered phospholipids represented ether acyl phosphatides. Differences in the lipid composition of rat vesicles described here and similar vesicles isolated from ram and boar spermatozoa (described previously) are discussed. The partitioning of the nitroxyl spin label 3-doxylheptane into vesicles isolated from rat and ram spermatozoa was assessed by electron paramagnetic resonance spectroscopy at temperatures between 4 C and 26 C; no difference in the response of the spin label in the two vesicle preparations was detected.

UI	MeSH Term	Description	Entries
D008297	Male		Males
D008563	Membrane Lipids	Lipids, predominantly phospholipids, cholesterol and small amounts of glycolipids found in membranes including cellular and intracellular membranes. These lipids may be arranged in bilayers in the membranes with integral proteins between the layers and peripheral proteins attached to the outside. Membrane lipids are required for active transport, several enzymatic activities and membrane formation.	Cell Membrane Lipid,Cell Membrane Lipids,Membrane Lipid,Lipid, Cell Membrane,Lipid, Membrane,Lipids, Cell Membrane,Lipids, Membrane,Membrane Lipid, Cell,Membrane Lipids, Cell
D008565	Membrane Proteins	Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.	Cell Membrane Protein,Cell Membrane Proteins,Cell Surface Protein,Cell Surface Proteins,Integral Membrane Proteins,Membrane-Associated Protein,Surface Protein,Surface Proteins,Integral Membrane Protein,Membrane Protein,Membrane-Associated Proteins,Membrane Associated Protein,Membrane Associated Proteins,Membrane Protein, Cell,Membrane Protein, Integral,Membrane Proteins, Integral,Protein, Cell Membrane,Protein, Cell Surface,Protein, Integral Membrane,Protein, Membrane,Protein, Membrane-Associated,Protein, Surface,Proteins, Cell Membrane,Proteins, Cell Surface,Proteins, Integral Membrane,Proteins, Membrane,Proteins, Membrane-Associated,Proteins, Surface,Surface Protein, Cell
D008854	Microscopy, Electron	Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.	Electron Microscopy
D010743	Phospholipids	Lipids containing one or more phosphate groups, particularly those derived from either glycerol (phosphoglycerides see GLYCEROPHOSPHOLIPIDS) or sphingosine (SPHINGOLIPIDS). They are polar lipids that are of great importance for the structure and function of cell membranes and are the most abundant of membrane lipids, although not stored in large amounts in the system.	Phosphatides,Phospholipid
D002458	Cell Fractionation	Techniques to partition various components of the cell into SUBCELLULAR FRACTIONS.	Cell Fractionations,Fractionation, Cell,Fractionations, Cell
D002462	Cell Membrane	The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.	Plasma Membrane,Cytoplasmic Membrane,Cell Membranes,Cytoplasmic Membranes,Membrane, Cell,Membrane, Cytoplasmic,Membrane, Plasma,Membranes, Cell,Membranes, Cytoplasmic,Membranes, Plasma,Plasma Membranes
D004822	Epididymis	The convoluted cordlike structure attached to the posterior of the TESTIS. Epididymis consists of the head (caput), the body (corpus), and the tail (cauda). A network of ducts leaving the testis joins into a common epididymal tubule proper which provides the transport, storage, and maturation of SPERMATOZOA.
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D012756	Sheep	Any of the ruminant mammals with curved horns in the genus Ovis, family Bovidae. They possess lachrymal grooves and interdigital glands, which are absent in GOATS.	Ovis,Sheep, Dall,Dall Sheep,Ovis dalli