A MODIFIED PROCEDURE FOR THE ANALYSIS OF MIXTURES OF TISSUE GANGLIOSIDES<sup>1</sup>

MacMillan, V.; Wherrett, John R.

doi:10.1111/j.1471-4159.1969.tb10361.x

Cited by 75 publications

(19 citation statements)

References 14 publications

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“…Protein and DNA were extracted from the brain tissue (22) and measured by the methods of Lowry et al (23) and Burton (24), respectively. Gangliosides were extracted by the method of Suzuki (25), resolved into major species by TLC, with Silica Gel H and the neuranimic acid (NANA) content determined by the resorcinol reaction (26). Cerebrosides, sulfatides, and ceramides were separated and purified by Florisil column chromatography and TLC with Silica Gel (27).…”

Section: Biochemical Analysesmentioning

confidence: 99%

Effect of Lead Intoxication on the Postnatal Growth of the Rat Nervous System

Krigman¹,

Hogan²

1974

Environmental Health Perspectives

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Lead encephalopathy was induced in developing Long-Evans rats by adding lead carbonate (4% w/w) to the diet of nursing mother immediately after delivery. The morphological and biochemical features of cerebral ontogenesis were studied in 30-dayold rats.By the 30th postnatal day, the overall effect of lead intoxication was retardation of brain growth. The mass of both the cerebral gray and white matter was appreciably reduced in the lead rats without any reduction in cell populations. While the neuronal population was preserved, the growth of neurons was reduced and their maturation retarded. The retarded neuronal growth was characterized by the limited proliferation of processes in the neuropil and by the reduction in the number of synapses per neuron. However, synaptogenesis was neither delayed nor perturbed but reduced by the limited development of neuronal dendritic fields. The myelination was altered and its cerebral content significantly reduced. The effect of lead on myelination was one of hypomyelination. The hypomyelination appears to be primarily related to retarded growth and maturation of the neuron and is not a reflection of a defect in the myelinating glia or a delay in the initiation of myelination. IntroductionThe vulnerability of the nervous system to the toxic effects of lead is most evident in the young. While the neural aspects of plumbism have been the subject of numerous studies, we still have much to learn about the extent of the effects of lead on the nervous system. This is particularly true for body lead burdens in children which have been previously considered nontoxic (1, 2). The nervous system of the young is obviously different from that of the adult. The episodic nature of the growth and maturation of the developing nervous system may make the young more vulnerable at selected periods (3) to a hostile environment than the adult.In an attempt to define the effects of lead on the developing nervous system, we have studied the model of lead encephalopathy May 1974 187 described by Pentschew and Garro (4). The model uses the nursing rat and is characterized by selected morphological features resembling those of clinical encephalopathy. This report is based upon quantitative morphometric and biochemical analyses of postnatal brain development and contains our findings on the effects of lead on neuronal growth and maturation, synaptogenesis, and myelination. The studies reported herein were limited to the 30th postnatal day. By this time the nervous system is highly developed: the brain weight is nearly threefourths that of an adult (5), neurogenesis has stopped (5, 6), synaptogenesis has nearly ceased (7,8), and the most active period of myelination is over (3); also, the pups are already weaned. Materials and MethodsA more detailed description of the animal model, morphometric procedures, and chemical analyses appears in our earlier reports (9, 10). Animal ModelLead intoxication was produced in the Long-Evans strain of rats by the procedure described by Pentschew and Garro (4). Immediate...

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Section: Biochemical Analysesmentioning

confidence: 99%

Effect of Lead Intoxication on the Postnatal Growth of the Rat Nervous System

Krigman¹,

Hogan²

1974

Environmental Health Perspectives

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“…This was followed by resorcinol assay directly on the positively identified spots which were scraped off the plate (12,13). In order to facilitate identification of spots containing small amounts of ganglioside a modification of the Suzuki (12, 13) procedure suggested by MacMillan and Wherrett (14) was employed. This was especially useful in estimating quantitative recoveries during the course of purification.…”

Section: Methodsmentioning

confidence: 99%

Studies on Human Platelet Gangliosides

Marcus¹,

Ullman²,

Safier³

1972

J. Clin. Invest.

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A B S T R A C T Gangliosides, glycosphingolipids which contain sialic acid, were studied in human platelets. They represented 0.5% of the platelet lipids and accounted for 6% of the total neuraminic acid content of platelets. Three major ganglioside fractions were identified and characterized. Ganglioside I was hematoside (G6) and comprised 92% of the platelet gangliosides. It contained glucose, galactose, and sialic acid in molar ratios of 1: 1: 1 and no hexosamnine. The imiajor fatty acid was behenate (22: 0). Ganglioside I w-as also identified in isolated platelet granules and membranes. Ganglioside II (5%) contained glucose, galactose. sialic acid, and hexosamines (molar ratios 1: 2: 1: 1). The hexosamines were glucosamine (72%) and galactosamine (28%). It was therefore designated as ganglioside lacto-N-neotetraose. Ganglioside III (2%) contained disialosyllactosyl ceramide (G3A) as well as two other gangliosides which could not be precisely characterized. Gangliosides I, II, and III were susceptible to the action of Clostridiuin perfringens neuraminidase as evidenced by full recovery of sialic acid in its free form after incubation. Neutral platelet glycolipids were qualitatively examined by thinlayer chromatography. The major component was lactosvI ceramide.Interactions of gangliosides I and III and serotonin-'4Cwere examined in an equilibrium dialysis system at 4°C. The gangliosides bound serotonin-'4C in relatively small quantities, wlhereas cointrol lipids w-ere negative. The binding was essentially tinchainged by reverse dialysis, ultracentrifugation anld stubsequen-t tlhin-laver chromatography. The results are comparable to the previously observed nonmetabolic interactions between wlhole platelets and serotonin in the cold. It is suggested that the orientation and specific distribtution of platelet membrane glycolipids may be important determinants of the unique surface properties of platelets.

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“…;C and their sugar composition determined by analyses by LBSA, protein and lipid were determined as described in the text. The percentage distribution of sialic acid in the individual gangliosides was measured by the method of MacMillan and Wherrett [27] ; the results are expressed as a percentage of the total sialic acid recoyered. The composition of the individual gangliosides of rabbit skeletal muscle has previously been reported by Lassaga et al [14].…”

Section: Methodsmentioning

confidence: 99%