Bulk Separation of Neuronal Cell Bodies and Glial Cells in the Absence of Added Digestive Enzymes

Sellinger, Otto Z.; Azcurra, Julio M.

doi:10.1007/978-1-4615-7751-5_1

Cited by 53 publications

(4 citation statements)

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“…Glial cells were isolated from whole brain (cerebrum and cerebellum) according to the method described by Sellinger & Azcurra (1974). Briefly, pooled brains were placed in beakers containing 7•5 % (w/v) polyvinylpyrrolidone and 10 mmol/l CaCl 2 at pH 4•7 and 25 8C.…”

Section: Isolation Of Glial Cells From Whole Brainmentioning

confidence: 99%

Maternal dietary 22: 6n-3 is more effective than 18: 3n-3 in increasing the 22: 6n-3 content in phospholipids of glial cells from neonatal rat brain

Bowen

Clandinin

2005

Br J Nutr

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One of the debates in infant nutrition concerns whether dietary 18: 3n-3 (linolenic acid) can provide for the accretion of 22: 6 n-3 (docosahexaenoic acid, DHA) in neonatal tissues. The objective of the present study was to determine whether low or high 18: 3 n-3 v. preformed 22: 6 n-3 in the maternal diet enabled a similar 22: 6 n-3 content in the phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and phosphatidylserine (PS) of glial cells from whole brain (cerebrum and cerebellum) of 2-week-old rat pups. At parturition, the dams were fed semi-purified diets containing either increasing amounts of 18: 3 n-3 (18: 2 n-6 to 18: 3 n-3 fatty acid ratio of 7·8: 1, 4·4: 1 or 1: 1), preformed DHA, or preformed 20: 4 n-6 (arachidonic acid)+DHA. During the first 2 weeks of life, the rat pups from the respective dams received only their dam's milk. The fatty acid composition of the pups' stomach contents (dam's milk) and phospholipids from glial cells were quantified. The 20: 4n-6 and 22: 6 n-3 content in the stomach from rat pups at 2 weeks of age reflected the fatty acid composition of the dam's diet. The 20: 4n-6 content of PE and PS in the glial cells was unaffected by maternal diet treatments. Preformed 22: 6 n-3 in the maternal diet increased the 22: 6 n-3 content of glial cell PE and PS compared with maternal diets providing an 18: 2n-6 to 18: 3 n-3 fatty acid ratio of 7·8: 1, 4·4: 1 or 1: 1 (P<0·0001). There was no significant difference in the 20: 4 n-6 and 22: 6 n-3 content of glial cell PC and PI among maternal diet treatments. It was concluded that maternal dietary 22: 6n-3 is more effective than low or high levels of maternal dietary 18: 3 n-3 at increasing the 22: 6 n-3 content in PE and PS of glial cells from the whole brain of rat pups at 2 weeks of age. The findings from the present study have important implications for human infants fed infant formulas that are devoid of 22: 6 n-3.

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Section: Isolation Of Glial Cells From Whole Brainmentioning

confidence: 99%

Maternal dietary 22: 6n-3 is more effective than 18: 3n-3 in increasing the 22: 6n-3 content in phospholipids of glial cells from neonatal rat brain

Bowen

Clandinin

2005

Br J Nutr

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“…Male pups were euthanized at day 14 after birth (n = 24 in each group). Brains (cerebrum plus cerebellum) were excised and placed in ice-cold 0.32 M sucrose as previously described [ 10 , 17 ]. Six brains from pups at day 14 were pooled together for glial cell separation.…”

Section: Methodsmentioning

confidence: 99%

“…Glial cells were purified from the whole brain (cerebrum plus cerebellum) by centrifugation as described in the literature [ 17 ]. Briefly, pooled brains were homogenized in 7.5% (w/v) polyvinylpyrrolidone and 10 mM CaCl 2 at pH 4.7 and 25°C.…”

Section: Methodsmentioning

confidence: 99%

Differential effect of maternal diet supplementation with α-Linolenic adcid or n-3 long-chain polyunsaturated fatty acids on glial cell phosphatidylethanolamine and phosphatidylserine fatty acid profile in neonate rat brains

Destaillats

Joffre²,

Acar³

et al. 2010

Nutr Metab (Lond)

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BackgroundDietary long-chain polyunsaturated fatty acids (LC-PUFA) are of crucial importance for the development of neural tissues. The aim of this study was to evaluate the impact of a dietary supplementation in n-3 fatty acids in female rats during gestation and lactation on fatty acid pattern in brain glial cells phosphatidylethanolamine (PE) and phosphatidylserine (PS) in the neonates.MethodsSprague-Dawley rats were fed during the whole gestation and lactation period with a diet containing either docosahexaenoic acid (DHA, 0.55%) and eicosapentaenoic acid (EPA, 0.75% of total fatty acids) or α-linolenic acid (ALA, 2.90%). At two weeks of age, gastric content and brain glial cell PE and PS of rat neonates were analyzed for their fatty acid and dimethylacetal (DMA) profile. Data were analyzed by bivariate and multivariate statistics.ResultsIn the neonates from the group fed with n-3 LC-PUFA, the DHA level in gastric content (+65%, P < 0.0001) and brain glial cell PE (+18%, P = 0.0001) and PS (+15%, P = 0.0009) were significantly increased compared to the ALA group. The filtered correlation analysis (P < 0.05) underlined that levels of dihomo-γ-linolenic acid (DGLA), DHA and n-3 docosapentaenoic acid (DPA) were negatively correlated with arachidonic acid (ARA) and n-6 DPA in PE of brain glial cells. No significant correlation between n-3 and n-6 LC-PUFA were found in the PS dataset. DMA level in PE was negatively correlated with n-6 DPA. DMA were found to occur in brain glial cell PS fraction; in this class DMA level was correlated negatively with DHA and positively with ARA.ConclusionThe present study confirms that early supplementation of maternal diet with n-3 fatty acids supplied as LC-PUFA is more efficient in increasing n-3 in brain glial cell PE and PS in the neonate than ALA. Negative correlation between n-6 DPA, a conventional marker of DHA deficiency, and DMA in PE suggests n-6 DPA that potentially be considered as a marker of tissue ethanolamine plasmalogen status. The combination of multivariate and bivariate statistics allowed to underline that the accretion pattern of n-3 LC-PUFA in PE and PS differ.

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“…Although the formation of SAM has been studied in great detail in yeast, E. coli and liver [4-7, 9-15, 18-21], only occasional reference to this process in brain exists, and, to our knowledge, the possibility of different molecular forms of ATP: L-methionine-S-adenosyltransferase in nerve versus glial cells [24] or between nerve cells differing in function [25] has not been ex plored. Since, however, the differential availability of SAM in a number of neuroanatomical regions of the rat brain [23] may in large part determine the méthylation and hence the specific metabolism of key molecules, such as, for instance, the biogenic amines, noradrenaline, dopamine, tryptamine and histamine, and of macromolecules under polyamine control, such as tRNA [1], it is important to study cerebral SAM formation in some detail.…”

Section: Discussionmentioning

confidence: 99%

A Comparison of the ATP: L-Methionine-S-Adenosyltransferase of Rat Cerebral Cortex and Cerebellum

Mc¹,

Oz²

1976

Enzyme

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The ATP: L-methionine-S-adenosyltransferase of rat cerebral cortex and cerebellum was found to be differentially responsive to solubilization by sodium deoxycholate. Furthermore, the cerebellar enzyme was markedly less sensitive to inactivation by deoxycholate and to storage at 4°C. The specific activity of the cerebellar enzyme was significantly higher and the two enzyme activities also exhibited differences in apparent K(m) values for L-methionine.

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Bulk Separation of Neuronal Cell Bodies and Glial Cells in the Absence of Added Digestive Enzymes

Cited by 53 publications

References 60 publications

Maternal dietary 22: 6n-3 is more effective than 18: 3n-3 in increasing the 22: 6n-3 content in phospholipids of glial cells from neonatal rat brain

Maternal dietary 22: 6n-3 is more effective than 18: 3n-3 in increasing the 22: 6n-3 content in phospholipids of glial cells from neonatal rat brain

Differential effect of maternal diet supplementation with α-Linolenic adcid or n-3 long-chain polyunsaturated fatty acids on glial cell phosphatidylethanolamine and phosphatidylserine fatty acid profile in neonate rat brains

A Comparison of the ATP: L-Methionine-S-Adenosyltransferase of Rat Cerebral Cortex and Cerebellum

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