Lipofuscin Pigment Research Articles

Ceroid lipofuscinosis in sheep. I. Bis(monoacylglycero)phosphate, dolichol, ubiquinone, phospholipids, fatty acids, and fluorescence in liver lipopigment lipids.

The ceroid lipofuscinoses are inherited lysosomal diseases of children characterized by a fluorescent lipopigment stored in a variety of tissues. Defects in lipid metabolism or the control of lipid peroxidation have been postulated to explain their pathogenesis. In the present study, lipopigment was isolated from the liver of sheep affected with ceroid lipofuscinosis. It was 70% protein, the rest being mainly lipids. These were only one-sixth as fluorescent as total liver lipids, but contained a number of fluorophors. None were major components of the lipopigment or the postulated fluorescent product of lipid peroxidation. Lipopigment lipids included the lysosomal marker bis(monoacylglycero)phosphate that contained 42.9% linoleate and 16.5% linolenate. Lipopigment neutral lipids were dolichol, dolichyl esters, ubiquinone, free fatty acids, and cholesterol, indicative of a lysosomal origin of the lipopigment. Phosphatidylcholine, phosphatidylinositol, phosphatidylserine, and phosphatidylethanolamine were present in proportions and with fatty acid profiles typical of lysosomes. No differences were found between the lipids of total control and affected livers, nor the fatty acid profiles of their phosphatidylcholine, phosphatidylethanolamine, or triglycerides. It is concluded that ovine ceroid lipofuscinosis is not a lipidosis, nor does the lipopigment arise from the abnormal peroxidation of lipids. Strong similarities between the lipopigment and the age pigment lipofuscin were noted.

Leydig cell hyperplasia.

The ultrastructure of Leydig cell hyperplasia is described. The hyperplastic Leydig cells differ morphologically from normal Leydig cells as well as from Leydig tumor cells. They have less smooth endoplasmic reticulum and less lipofuscin pigment in their cytoplasm, and the nuclear membrane shows more irregular indentations than normal Leydig cells. It is suggested that electron microscopy might be useful in the differential diagnosis between Leydig cell hyperplasia, Leydig cell tumors, and normal Leydig cells.

Chapter 12 Ratio of pyramidal cells versus non-pyramidal cells in the human frontal isocortex and changes in ratio with ageing and Alzheimer's disease

This chapter presents a study that assesses the percentages of both the pyramidal neurons and the non-pyramidal cells in various layers of a defined prefrontal isocortical area of the human adult and it has the ultimate goal of examining whether changes of these percentages occur with ageing and in Alzheimer's disease. The neuronal constituents of the mammalian telencephalic cortex can generally be classified as either pyramidal neurons or non-pyramidal neurons. In general, pyramidal cells and the modified forms of pyramidal cells can be considered as projection neurons, while most non-pyramidal cells can be referred to as local circuit neurons. Lipofuscin pigment granules can frequently be encountered in nerve cells of the human adult. The size, shape, stainability, and pattern of distribution of these granules can serve as characteristics for the differentiation of various neuronal types forming the human brain. Pigmented nerve cells show a slight increase in the number of lipofuscin granules with age, but this does not change the typical pattern of pigmentation. The pigmentation of pyramidal cells and modified pyramidal cells differs considerably from that seen in the nonpyramidal neurons. After the demonstration of the correspondence of the pigment characteristics and the nerve cell type as seen in the Golgi preparation, a combined pigment, Nissl, preparation allow to quantitatively analyze the proportions of the various types of cortical constituents at light microscopical level and enables to study their pattern of distribution throughout various cortical layers.

Effect of Dietary Selenite on Hepatic Organic Solvent-Soluble Lipofuscin Pigments

Supplementation of selenium as sodium selenite results in an increase in hepatic organic solvent-soluble lipofuscin pigments, the metabolic end products of lipid peroxidation. Weanling mice fed a basal diet containing 0.05 ppm selenium had a significant increase in hepatic organic solvent-soluble lipofuscin pigments and glutathione peroxidase activity following supplementation of an additional 0.1 ppm selenium as sodium selenite from 5 to 9 months of age. Normal levels of vitamin E (30 mg/kg) were insufficient to protect against the oxidative effect of this increased dose of selenite. However, 10 times the normal level of vitamin E markedly suppressed this oxidative effect.

Foveomacular Vitelliform Dystrophy, Adult Type: A Clinicopathologic Study Including Electron Microscopic Observations

Foveomacular vitelliform dystrophy, adult type (FVDAT), is a dystrophy of the retinal pigment epithelium (RPE) characterized in part by autosomal dominant inheritance, midlife onset, mild visual loss, and various patterns of foveal yellow deposits located at the level of the pigment epithelium. Detailed pathologic studies of two postmortem eyes from a 61-year-old woman with documented FVDAT disclosed striking abnormalities involving mainly the foveal RPE and overlying photoreceptors. By light microscopy, marked focal atrophy of the pigment epithelial cells in the foveolar area was bordered by hypertrophic RPE. Interposed between the atrophic RPE and Bruch's membrane were scattered eosinophilic, fusiform collagenous plaques. The sensory retina overlying the deranged RPE displayed marked atrophy of the outer nuclear layer with loss of photoreceptor inner and outer segments. Pigment-laden macrophages containing periodic acid -Schiff-positive material had migrated into the atrophic, outer sensory retina. Ultraviolet fluorescent microscopy demonstrated massive accumulation of lipofuscin pigment within the macular RPE as well as within macrophages in the atrophic outer retina. By scanning electron microscopy, a confluent area of flattened, atrophic retinal pigment epithelial cells was rimmed by taller, hypertrophic RPE cells..By transmission electron microscopy, the pigment epithelial cells contained myriad lipofuscin granules. Bruch's membrane and the intercapillary pillars were slightly thickened and the choriocapillaris was patent. The possible interrelationships between FVDAT and the other pattern dystrophies of the RPE are discussed.

ChemInform Abstract: STUDIES ON PEROXIDIZED LIPIDS. VI. FLUORESCENT PRODUCTS DERIVED FROM THE REACTION OF PRIMARY AMINES, MALONALDEHYDE AND MONOFUNCTIONAL ALDEHYDES

AbstractMalondialdehyd (II) liefert mit den primären Aminen (III) die 4‐Methyldihydropyridindicarbaldehyde (I), die Cokondensation der Aldehyde (IV) mit (II) und (III) führt zu den 4‐substituierten Dihydropyridindicarbaldehyden (V).

Population age structure in males and juveniles of the Antarctic Krill,Euphausia superba Dana

Further results are reported on the age structure of populations of the Antarctic krill, using the age pigment lipofuscin. Study of the males shows a five-year adult life span as in females in an earlier study. Only a single age group was found in the juvenile stage.

The rôle of copper level in the formation of neuronal lipofuscin in the spinal ganglia of Torpedo m

Copper, a marine pollutant of the Gulf of Naples, appears to be involved in the formation of lipofuscin in neuronal cells of Torpedo m. Lipofuscin pigment, a wear and tear pigment, is considered to be a marker of cell damage—in particular in cells such as neurons which are post-mitotic cells which reveal the life history of an an individual. Samples of Torpedo m. were collected from two sites with different levels of pollution. Other animals were kept for 60 days in a closed circuit seawater tank enriched with CuCl 2. In addition to cells of the central nervous system (CNS), other organs were sampled for copper content. Detailed quantification of lipofuscin was made, at the electronmicroscopic level, of spinal ganglia cells and cells of the electric lobe, revealing the high content of lipofuscin in the spinal ganglia neurons of fish from a heavily polluted area and a further increase of lipofuscin in those neurons of animals kept in artificially high copper-containing seawater. The general implications of this pollution for the Gulf of Naples are discussed.

Ratio of pyramidal cells versus non-pyramidal cells in sector CA1 of the human Ammon's horn

Combined Golgi/pigment studies revealed that pyramidal neurons and non-pyramidal cells of the Ammon's horn of the human adult can be distinguished from each other by their characteristic lipofuscin pigment deposits. In sector CA1, both the typical pyramidal neurons and the modified forms of pyramidal cells contain a modest amount of fine lipofuscin granules while non-pyramidal cells are either pigment-laden or devoid of lipofuscin deposits. Strips running through the whole depth of the pyramidal cell layer and the stratum oriens of CA1 were examined and all nucleolated nerve cells present within these strips were classified and counted (16 brains, age range from 28 to 69 years). Of the 18,510 neurons classified, 16,765 were pyramidal cells, including their modified versions, and 1,745 were non-pyramidal cells. The pyramidal cells, accordingly, were intermixed with 9.4 +/- 1.0% non-pyramidal neurons. The data presented provide a basis for investigation of the aging and diseased human brain.

Studies on peroxidized lipids. VI. Fluorescent products derived from the reaction of primary amines, malonaldehyde and monofunctional aldehydes

AbstractMonofunctional aldehydes such as acetaldehyde,n‐propylaldehyde,n‐butylaldehyde,n‐hexylaldehyde,n‐heptylaldehyde and benzaldehyde affected the reaction between primary amines and malonaldehyde. While the reaction of primary amines and malonaldehyde at pH 7 produced fluorescent 4‐methyl‐1,4‐dihydropyridine‐3,5‐dicarbaldehydesIa‐f, the reaction of the primary amines, malonaldehyde and the aldehydes listed above gave fluorescent 4‐substituted 1,4‐dihydropyridine‐3,5‐dicarbaldehydesIIa‐j. The primary amines used for this reaction included alkylamines, amino acids and alkanolamines. The optimal ratio of the amine, malonaldehyde and the aldehyde was 1:2:1–2, at which compoundsII were produced quantitatively. Peroxidized lipids which may contain malonaldehyde and other aldehydes could react with the primary amines to produce highly fluorescentII. Fluorescence spectra ofII showed excitation maxima at 386–403 nm and emission maxima at 444–465 nm in phosphate similar to those ofI. The spectra of these 1,4‐dihydropyridinesI andII were roughly similar to those of lipofuscin pigment, but they exhibited different characteristics in acid and alkaline media from those of lipofuscin pigment. CompoundsII may be useful as model compounds to elucidate the chemical structure of lipofuscin pigment.

The human placenta does not contain lipofuscin pigment

Autofluorescent granules are present in the villous syncytiotrophoblast of the human placenta, most prominently during the first trimester. These granules differ in both size and hue from classical lipofuscin granules, have different staining characteristics, and are shown by electron microscopy to be syncytial lipid droplets. It is concluded that previous reports of the presence of lipofuscin pigment in the placenta are unfounded.

Effect of Selenite, Vitamin E and N,N′-Diphenyl-p-Phenylenediamine on Liver Organic Solvent-Soluble Lipofuscin Pigments in Mice

The present experiment was designed to investigate the effect of selenium (Se) supplementation, as sodium selenite, on organic solvent-soluble lipofuscin pigment (OLP) accumulation and glutathione peroxidase (GSH-Px) activity in the livers of mice fed varying levels of vitamin E or N,N′-diphenyl-p-phenylenediamine (DPPD). Four groups of 16 female, weanling mice each were fed either a vitamin E-deficient diet, a diet supplemented with 30 mg/kg or 300 mg/kg vitamin E (as RRR-α-tocopheryl acetate), or a diet supplemented with 30 mg/kg DPPD. Each diet contained 0.05 ppm Se. At 5 months of age, eight animals from each dietary group were supplemented with an additional 0.1 ppm Se, as sodium selenite, in their drinking water. The remaining animals were fed their original diets through the 9-month experimental period. Selenite supplementation resulted in a significant increase in OLP concentration and GSH-Px activity in the liver of mice fed vitamin E- or DPPD-supplemented diets. Normal levels of vitamin E and DPPD (30 mg/kg) were not sufficient to protect against the oxidative effects of selenite; however, 10 times the normal level of vitamin E (300 mg/kg) markedly suppressed this oxidative effect.

The neuromuscular features of acromegaly: a clinical and pathological study.

A study of the neuromuscular features of acromegaly was performed in six patients. Clinical assessment was supplemented by quadriceps force measurements, plasma creatine kinase (CK) activities, electromyography (EMG) and nerve conduction studies. Muscle mass was measured by urinary creatinine/height indices (CHI) and cross sectional area (CSA) of thighs and calves on computed tomography. Quadriceps force/unit cross sectional area was derived. Needle biopsies of vastus lateralis were studied by histochemical and ultrastructural methods. Mean fibre area (MFA) and fibre type proportions were measured. Most of the subjects studied had muscle pain and proximal muscle weakness confirmed by quadriceps force measurements. This occurred in the absence of muscle wasting, as shown by cross sectional area measurements and normal or raised creatinine/height indices. "Myopathic" features were demonstrated by needle biopsy in half the patients and occasionally by electromyography and raised plasma creatine kinase activity. Abnormalities on needle biopsy included variation in fibre size, type 2 fibre atrophy and large type 1 MFA relative to type 2 MFA. Electronmicroscopy showed the non-specific findings of increased glycogen accumulation, excess lipofuscin pigment and myofilament loss.

Identification and characterization of non-dividing cell populations in phase II cultures of human glial cells

It has been reported that cultures of normal cells in phase II contain a non-multiplying cell population, the size of which increases with passage number. In phase II cultures of normal human glial cells we have found two subpopulations of non-proliferating cells, one of which has a characteristic morphology, and differs from the actively dividing cells in a number of respects: (1) they are larger although of various sizes and are well spread over a very large substratum area: (2) they contain a great number of granules showing acid phosphatase activity, being heavy metal positive and displaying the characteristic natural fluorescence of lipofuscin pigment; (3) they frequently contained a central somewhat irregular nucleus with various numbers of darkly straining nucleolar-like structures. Cytophotometric nuclear DNA measurements of the described “large” cell population show a decreased proportion of diploid cells as compared to their smaller sister cells. Moreover, with increasing passage number, the DNA values for large cells shift towards higher ploidy levels resulting in a scattered aneuploid pattern in the oldest passage. This “large” cell subpopulation consists of between 2% and 3% of all passages and become greatly decreased following subcultivation. The other subpopulation of non-dividing cells is generally morphologically similar to the dividers, increases in size with passage number and is the more important in the phase III phenomenon.

Changes in nerve cells of the nucleus basalis of Meynert in Alzheimer's disease and their relationship to ageing and to the accumulation of lipofuscin pigment

The number of nerve cells in the nucleus basalis of Meynert in normally aged persons is reduced by 30% by 90 years of age and cytoplasmic RNA content and nucleolar volume by about 20%. In Alzheimer's disease the changes are greatly exacerbated, with the cell number being depleted by a further 60% and cytoplasmic RNA content and nucleolar volume by an extra 35%. Moreover, younger patients with Alzheimer's disease show a much greater difference from controls of the same age than do older patients; indeed, by 90 years of age the levels of these changes are similar in Alzheimer's disease and in old age alone. Lipofuscin content is increased to a similar extent with age in both Alzheimer's disease and control patients and was associated with loss of cytoplasmic RNA content and nucleolar volume in both groups. It is suggested that mechanisms which result in the accumulation of this pigment during life also lower the capability of cells of the nucleus basalis of Meynert to withstand disease pathogens, leading to a certain degree of change in old age alone and one which is other persons may be compounded by secondary factors giving the extreme degeneration of Alzheimer's disease.

Neuronal types in the neocortex-dependent lateral territory of the human thalamus. A Golgi-pigment study.

Nerve cell types of the neocortex-dependent nuclei of the human thalamus were investigated with the use of a transparent Golgi technique, that allows one to study not only the peculiarities of the cell processes, but also the marking characteristics of the intraneuronal lipofuscin pigment deposits. Three principal types of neurons have been distinguished: Type I is a medium-sized to large neuron with a profusely radiating dendrite system. Numerous large vacuolated lipofuscin granules are contained in one pole of the cell body. Type II is a small to medium-sized neuron with a few sparsely branching dendrites. Small and intensely stained pigment granules are dispersed within the cell body. Type III is a medium-sized to large neuron with only a few thick and almost unbranched dendrites devoid of spiny appendages. The dendrites extend over long distances. The cell body is devoid of lipofuscin granules.

Ultrastructure of the testis in Klinefelter's syndrome.

The ultrastructure of the testis in 5 cases of Klinefelter's syndrome was studied. In 3 cases there was complete hyalinization of the seminiferous tubules. In one case Sertoli cells were present, and in another case focal areas of spermatogenesis were seen in the tubules. Hyperplasia of the Leydig cells was found in every case. The Leydig cells had usually abundant smooth endoplasmic reticulum and lipofuscin pigment in the cytoplasm. This suggested that the function of the Leydig cells was normal. The Sertoli cells contained lipid droplets and glycogen-filled vacuoles in the cytoplasm.

Studies on peroxidized lipids: Fluorescent products derived from the reaction of unsaturated fatty acids and methylamine.

Unsaturated fatty acids such as linoleic, linolenic and arachidonic acids produced a complex mixture of fluorescent substances with an excitation maximum at 355-370nm and an emission maximum at 420-440nm by reaction with methylamine at pH 7.5 and 37°C. The excitation and emission maxima of the products shifted to higher wavelength with increasing unsaturation of the fatty acid, and were similar to those of lipofuscin pigments. The fluorescence characteristics of the products indicated that they were different in structure or composition with different fatty acids, and the major fluorescent products were neither 1-substituted-4-methyl-1, 4-dihydropyridine-3, 5-dicarbaldehydes [Kikugawa et al., Chem. Pharm. Bull., 29, 1423 (1981)] nor the conjugated Schiff bases [Chio and Tappel, Biochemistry, 8, 2821 (1969)] which were produced by reaction of malonaldehyde (MDA) with primary amines. Both thin-layer and high performance liquid chromatographies suggested that thiobarbituric acid-reactive substances other than MDA contributed to the formation of the fluorescent substances.

Relationship between pigment accumulation and age in Alzheimer's disease and Down syndrome.

The amount of lipofuscin pigment within nerve cells of the nucleus basalis of Meynert and that of melanin pigment with nerve cells of locus ceruleus was measured in seven patients with Down syndrome, in 22 patients with Alzheimer's disease and in 18 controls ranging from 30 to 88 years of age. No significant differences in amount of either pigment in these cells at any age were noted between the patient groups and their age controls.

A preliminary report on hepatic pigmentation in Falkland Islands sheep.

Pigmentation of hepatocytes and Kupffer's cells with lipofuscin and melanin-like pigment is described. The occurrence of pigmented reticulo-endothelial cells in the lungs and lymphoid tissue was also noted. No discernible impairment of function accompanied the pigmentation.