Plasmalogen Content Research Articles

Phospholipids of bovine spermatozoa and seminal plasma.

Summary. Spermatozoal and seminal plasma lipids of fourteen individual bulls were separated by column chromatography into neutral lipid and several phospholipid fractions. Elution progress was monitored by thin-layer chromatography. Each phospholipid constituent was determined by phosphorus analysis. Total lipid, cholesterol and plasmalogen contents were determined. The fatty acids and aldehydes of the choline and ethanolamine phosphatide fractions were analysed by gasliquid chromatography. The spermatozoal phospholipid comprised 35·6% phosphatidyl choline, 28% phosphatidal choline, 20% phosphatidyl ethanolamine, 7·2% phosphatidal ethanolamine and 9·1% sphingomyelin. The seminal plasma phospholipid comprised 30% phosphatidyl choline, 23·6% phosphatidal choline, 10·5% phosphatidyl ethanolamine, 16·3% phosphatidal ethanolamine, 14·1% sphingomyelin and 5·4% polyglycerol phosphatide. Myristaldehyde and palmitaldehyde were the only aldehydes identified in the choline and ethanolamine phosphatide fractions. Docosahexaenoic acid constituted a large portion of the fatty acids of spermatozoal choline phosphatide.

Changes in plasmalogen content of human heart and skeletal muscle with age and development.

THE proportion of plasmalogen to total phospholipid in mammalian heart muscle seems to depend on both the age and species of the animal, with the choline plasmalogen (glycerol-2-acyl-1-alkenyl-3-phosphorylcholine) showing the most marked variations1–3. Species differences in the plasmalogen content of skeletal muscle also seem to be considerable and on the basis of analyses of monkey muscle, primate skeletal muscle has been suggested to be considerably richer in plasmalogens than tissue from other species, such as the rabbit4. Some of our analyses of mature and immature human muscle have shown that maturational changes, somewhat similar to those occurring in heart muscle, also take place in skeletal muscle. In addition they suggest that the plasmalogen content of mature human skeletal muscle is similar to that of monkey muscle.

Determination of plasmalogens after treating with a 2,4‐dinitrophenylhydrazine‐phosphoric acid reagent

A method for the quantitative determination of plasmalogens is described. Glycerophosphatide fractions from animal and plant sources were treated with a 2,4-dinitrophenylhydrazine-phosphoric acid reagent. The plasmalogen content was estimated by two different procedures after a single treatment with reagent: a) by determining the phosphorus content of diacyl phospholipid and (2-acyl) lysophospholipid separated by thin-layer chromatography, b) by spectrophotometric determination of the 2,4-dinitrophenylhydrazones and phosphorus assay of the original untreated sample. The total plasmalogen content of an unfractionated extract may be determined easily by the latter procedure.The method described gives results comparable with those from the p-nitrophenylhydrazine method and iodination procedure. The usefulness of this 2,4-dinitrophenylhydrazine-phosphoric acid treatment for further structural analyses of plasmalogens is discussed.

Lipid composition and metabolism in testicular and ejaculated ram spermatozoa.

1. Spermatozoa collected directly from the testis of the conscious ram contain 25% more phospholipid than ejaculated spermatozoa. The concentration of lecithin, phosphatidylethanolamine and ethanolamine plasmalogen was greater in testicular spermatozoa; little difference was observed in choline plasmalogen. Both types of spermatozoa had significant amounts of cardiolipin and alkyl ether phospholipid. 2. The fatty acids in the phospholipid extracted from testicular spermatozoa have a very high content of palmitic acid. The phospholipids of ejaculated spermatozoa contained less palmitic acid, but more myristic acid. 3. Ejaculated spermatozoa contained less acyl ester and cholesterol. It is suggested that lipids are a source of substrate for spermatozoa during their passage through the epididymis. 4. Testicular spermatozoa when incubated with [U-(14)C]glucose incorporated more radioactivity into the glycerol part of the phospholipid and neutral lipid fractions than did ejaculated cells. The distribution of radioactivity in the individual phospholipids and neutral lipids was similar for both cell types. No radioactivity was detected in choline plasmalogen, which accounted for approx. 40% of the total phospholipid. 5. Testicular spermatozoa incorporated more radioactivity from glucose into formate than into acetate, whereas a higher proportion of radioactivity was found in acetate in ejaculated cells. 6. The implications of these lipid changes in the process of spermatozoal maturation are discussed.

Phospholipid and cholesterol content of epididymal and ejaculated ram spermatozoa and seminal plasma in relation to cold shock.

The concentration of total phospholipid, phosphatidylcholine, phosphatidylethanolamine, and choline plasmalogen in spermatozoa from the caput epididymis of the ram was higher than in spermatozoa from the cauda. The phospholipid composition of spermatozoa from the latter region of the epididymis more closely resembled ejaculated spermatozoa.

Quantitative differences in phospholipids of ejaculated spermatozoa and spermatozoa from three levels of the epididymis of the boar.

Summary. Five major phospholipids of ejaculated and epididymal porcine spermatozoa were separated and quantitatively determined by thin-layer chromatography. These were in the order of decreasing concentration, phosphatidyl choline, phosphatidyl ethanolamine, ethanolamine plasmalogen, sphingomyelin and choline plasmalogen. Phosphatidyl serine, lysolecithin and possibly phosphatidyl inositol were also present, but in low concentrations. The concentration of phosphatidyl choline and phosphatidyl ethanolamine in spermatozoa progressively decreased from the head to the tail of the epididymis, and was lowest in ejaculated semen. The plasmalogen content of spermatozoa from all levels of the reproductive tract of the boar was low and varied very slightly.

Changes in lipid composition of the maturing rat testis.

1. The lipids and fatty acids of the lipids of testes of rats aged 4 weeks to 6 months were separated and analysed. 2. A decrease in concentration of triglyceride was noted, but there was no significant change in the concentration of phospholipids, plasmalogen or cholesterol during this time. 3. There were no significant differences in the total lipid concentration of palmitic acid, stearic acid, linoleic acid, arachidonic acid and docosatetraenoic acid between the various age groups. 4. A decrease in the concentration of oleic acid in the phosphatide and triglyceride fractions and an increase in the concentration of docosapentaenoic acid (characterized as the Delta4,7,10,13,16-isomer) in phosphatides but not in triglyceride were observed during the maturation period. 5. Histological studies indicated that the lipid changes occurred at the same time as the appearance and maturation of the spermatids.

PLASMALOGEN AND GLYCEROL ETHER CONCENTRATIONS IN NORMAL AND ATHEROSCLEROTIC AORTIC TISSUE.

PLASMALOGEN AND GLYCEROL ETHER CONCENTRATIONS IN NORMAL AND ATHEROSCLEROTIC AORTIC TISSUE.

STUDIES IN THE HISTOCHEMISTRY OF PLASMALOGENS: I. THE EFFECT OF FORMALIN AND ACROLEIN FIXATION ON THE PLASMALOGENS OF ADRENAL AND BRAIN

Formalin containing fixatives were found to cause a rapid drop in extractable plasmalogens of steer adrenal cortex and medulla and rat brain to 50-60% of the initial value in 4-6 hours. This rapid destructive reaction was followed by a slow decrease so that for several days the plasmalogen levels remained relatively constant. An acrolein containing fixative, although not evaluated with adrenal, showed similar behavior with rat brain. Control experiments show the fixative aldehyde is solely responsible for this plasmalogen loss. This decrease is not paralleled by a similar decrease in extractable total lipid or total phospholipid. It is believed that this decrease represents an actual destruction of the enol-ether bond by chemical reaction with the fixative aldehyde. Although the nature of this reaction is unknown, model experiments show cephalin plasmalogen to be much more labile than choline plasmalogens to formaldehyde, although all the cephalin plasmalogens are not destroyed during the fast reaction in tissue. These experiments also show that steer adrenal cortex and medulla have the same plasmalogen content and that the plasmalogen in either part of the gland responds the same to fixation, even though plasmal staining would indicate that the medulla is richer in plasmalogen. After a period of fixation sufficient to render the plasmal reaction negative in both adrenal cortex and medulla and in rat brain there is still at least 35-50% of the initial plasmalogen present. The intensity of the plasmal stain appears to parallel the concentration of the fixative labile plasmalogen. In these particular tissues there was no suggestion of an increase in free aldehyde over plasmalogen until the tenth day of fixation.

HISTOCHEMICAL STUDY ON THE CYTOPLASMIC CONSTITUENTS OF KIDNEY OF TOAD (BUFO MELANOSTICUS) DURING ACUTE DEHYDRATION

1. A condition of reduced glomerular filtration and increased tubular reabsorption was produced experimentally in toads by the process of acute dehydration. 2. In such a condition there was an interesting change, hitherto unrecorded, in acid phosphatase and plasmalogen content of the kidney. There was a fall in plasmalogen and a rise in acid phosphatase in kidney glomeruli during dehydration, and opposite results were observed in the tubules. 3. It may be concluded that plasmalogen is related to the functional activity of the kidney; in the case of acid phosphatase, a non-functional accumulation has been observed. 4. There was also a rise in succinic acid dehydrogenase and esterase in the functioning kidney tubules during dehydration.

Comparative studies on the phosphatides of normal rat liver and primary hepatoma.

A low phosphatide content was found in primary hepatoma of rat when compared with normal rat liver. Chromatographic analyses did not reveal, however, remarkable differences in the phosphatide composition of the normal and neoplastic tissue. The ratio of the 32P incorporation in vitro into phosphatidylcholine and phosphatidylethanolamine was lower in normal rat liver than in primary liver tumour. Gas chromatographic analyses of fatty acids performed on total lipid and its different components disclosed also notable differences between hepatoma and normal liver. A significant difference was observed in the stearic acid-oleic acid proportions between phosphatides of rat liver and primary hepatoma. The plasmalogen content of hepatoma was relatively rather high. The comparative analysis of phosphatides was extended to cell fractions obtained by differential centrifugation and to a mitochondrial membrane fragment. Within one tissue the fatty acid composition appeared to be characteristic for the lipid component and was independent of the cell fraction. A similar 32P pattern of the phosphatides from all cell fractions of one tissue was found after an incubation of slices with radioactive phosphate. Consequently the observed differences between phosphatides of normal rat liver and hepatoma are distributed throughout the cells.

Studies on the plasmalogens of nervous tissue

The concentration and distribution of plasmalogens in various regions of the human nervous system and in nervous and other tissues of some animal species have been investigated. Only traces of choline plasmalogen were detected in any region of the human or animal nervous systems examined. About 50% of the ethanolamine phosphatide of grey matter and probably all or nearly all that of white matter and spinal cord is present as plasmalogen.

The structure of plasmalogens IV. Lipids in normal and neoplastic tissues of man and in normal tissues of rabbit and rat

1. 1. Total lipid extracts of rabbit tissues (9 normal), rat tissues (9 normal, 1 cancerous), and human tissues (7 normal, 10 neoplastic) have been analyzed for their plasmalogen content by 2 independent methods: formation of the aldehyde p-nitrophenylhydrazone and addition of iodine under conditions specific for α,β-unsaturated ethers. 2. 2. The results agree with an average variation of less than 10%, indicating that almost all plasmalogens in mammalian organs have, as a common structural feature, an α,β-unsaturated ether linkage in a relatively long fatty chain. 3. 3. It is suggested that as the plasmalogen-lipid phosphorus ratio for a given organ is relatively constant, this ratio may be a more significant criterion than plasmalogen content for relating plasmalogen changes to organ function.

Histochemical studies on the dependence on ovarial function of the plasmalogen content of the genital tract in the white mouse

Der Plasmalogengehalt in den Epithelien des Genitaltractus der weisen Maus ist weitgehend abhangig von der Ovarialfunktion und stimmt besonders in der Uterusmucosa und in der Scheidenschleimhaut mit der Follikulinkurve uberein.