Isolation of oligodendroglial cells from young and adult whole rat brains using an in situ generated percoll density gradient

Mattera, Liliana N. Berti; Larocca, Jorge N.; Iraldi, Amanda Pellegrino de; Pasquini, Juana M.; Soto, Eduardo F.

doi:10.1016/0197-0186(84)90024-x

Cited by 46 publications

(29 citation statements)

References 19 publications

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“…Flow cytometric analysis of propidium iodide-stained oligodendrocytes from 15-day-old control (A) and hyperthyroid (B) rat brain. Oligodendroglial cells were isolated on a Percoll density gradient (Berti-Mattera et al, 1984), stained with propidium iodide and analyzed using a FAC-SCAN flow cytometer (for details see Materials and Methods). The percentage of hypodiploid-apoptotic oligodendrocytes was calculated using the Lysis II research software program.…”

Section: Discussionmentioning

confidence: 99%

“…One representative experiment of 12 different preparations is shown. (12) 12 Ϯ 4 3 1 Ϯ 5 *Oligodendroglial cells were isolated from total rat brain at different ages, using a Percoll autogenerated density gradient according to Berti-Mattera et al (1984). …”

Section: Discussionmentioning

confidence: 99%

“…Oligodendroglial cells were isolated from hyperthyroid and control rats of 5, 10, and 15 days of age on a Percoll density gradient, according to the method developed in our laboratory by Berti-Mattera et al (1984). The oligodendroglial cell population was immunostained with an antibody against O 4 .…”

Section: Evaluation Of Apoptosis By Flow Cytometrymentioning

confidence: 99%

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Sustained neonatal hyperthyroidism in the rat affects myelination in the central nervous system

et al. 1998

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We have carried out a study of the effects of sustained neonatal hyperthyroidism on myelin and on the oligodendroglial cells, in an effort to obtain further insight into the molecular mechanisms underlying the action of thyroid hormones on the central nervous system (CNS). Expression of the mRNAs of myelin basic protein (MBP) myelin proteolipid protein (PLP), 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase), transferrin, and c-Jun was investigated in 10- and 17-day-old normal and hyperthyroid rats, using Northern blot analysis. At 10 days of age, the levels of all the explored mRNAs were markedly higher in the experimental animals. The mRNA of transferrin showed a ninefold increase over control values, suggesting the possibility that this putative trophic factor might act as one of the mediators in the action of thyroid hormones. At 17 days of age on the other hand, the levels of all the mRNAs decreased markedly, reaching values below control, except for c-Jun, which remained higher than in normals. At 70 days of age, hyperthyroid rats showed clear evidence of myelin deficit, in agreement with previous results of our laboratories (Pasquini et al.: J Neurochem 57: Suppl S124, 1991). Immunocytochemistry of 70-day-old rat brain tissue sections showed a substantial reduction in the amount of MBP-reacting structures and a marked decrease in the number of oligodendroglial cells. Although the above-mentioned results could be the consequence, as proposed by Barres et al. (Development 120:1097-1108, 1994) and Baas et al. (Glia 19:324-332, 1997) of a premature arrest in oligodendroglial cell proliferation followed by early differentiation, the persistent high levels of expression of c-Jun, together with the dramatic decrease in the number of oligodendrocytes, suggested the possibility that prolonged hyperthyroidism could activate apoptotic mechanisms in the myelin forming cells. Using propidium iodide-labeled isolated oligodendroglial cells, we found, by flow cytometry, a significant increase in the number of apoptotic/hypo-diploid propidium iodide-positive cells. These results indicate that one of the actions of sustained levels of thyroid hormones in the neonate rat is to increase oligodendroglial cell death by apoptosis.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Sustained neonatal hyperthyroidism in the rat affects myelination in the central nervous system

et al. 1998

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“…Studies done by us, using oligodendrocytes isolated by the method developed by Berti-Mattera et al [54] in our laboratory, showed that microperoxisomes peaked at the period of myelin forma tion [55], It has been shown that T4 treatment increases the number of peroxisomes in the kidney and liver [56], In order to determine whether there was any correlation between accelerated myelin formation produced by thy roid honnones and the amount of microperoxisomes in the CNS, we studied the chemical composition of iso lated myelin as well as the levels of two peroxisomal marker enzymes, catalase and acyl-CoA dihydroxyacetone phosphate acyltransferase, in control and hyperthy roid animals [281. T3-treated animals appeared to initiate myelinogenesis earlier than controls.…”

Section: Effects Of Excess Thyroid Hormone On the Maturation Of Rat Bmentioning

confidence: 99%

Thyroid Hormones and the Central Nervous System

Pasquini

Adamo²

1994

Dev Neurosci

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Thyroid hormones have a significant influence on the development and maturation of the central nervous system. Among their actions, T₃ and T₄ have effects on the differentiation of various cell types in the rat brain and cerebellum as well as on the process of myelination. Recently, several investigators have shown effects of thyroid hormones on myelin protein gene expression. Thyroid hormones seem to have a regulatory role with regard to life span. Hyperthyroid animals appear to have a shorter life and, at advanced age, show a myelin deficit. This may be due to the damage produced by the oxidative stress generated by an excess of thyroid hormones.

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“…OLGcs were isolated from 4-and 10-day-old rat brain, using a Percoll density gradient, according to the method described in our laboratories by Berti-Mattera et al (1984) with slight modifications (C. Sato-Bigbee, personal communication). Briefly, the brains were minced and dissociated in Ca 2ϩ /Mg 2ϩ -free Hanks' balanced salt solution (HBSS), 25 mM HEPES (pH 7.2), 1 mg/ml glucose, 0.01 mg/ml DNase I, and trypsin 0.1%.…”

Section: Isolation and Culture Of Olgcsmentioning

confidence: 99%

Differential effects of apotransferrin on two populations of oligodendroglial cells

Paez

Soto

Pasquini

2003

Glia

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In the central nervous system (CNS), apotransferrin (aTf) is produced by oligodendroglial cells (OLGcs), and aTf is essential for cell survival. We previously demonstrated that a single intracranial injection of aTf in 3-day-old rats accelerates differentiation of OLGc and that aTf acts at early stages of development on certain populations of OLGcs, promoting accelerated maturation, with no effect on late markers of cell differentiation. The objective of the present study was to analyze OLGc maturation at two different stages of rat development, 4 and 10 days of age, in OLGcs isolated from the brain after intracranial injection of aTf at 3 days of age, and to explore the in vitro effect of aTf added to cultures of OLGc isolated from aTf-injected and control brains. The maturational cell stages were identified by immunocytochemistry with different OLGc markers and by analysis of their morphological complexity. The OLGcs isolated from 4- and 10-day-old animals intracranially injected with aTf were more differentiated than control cells. Treatment with aTf of the cultures of OLGcs that were isolated from 4-day-old saline-injected control animals induced their differentiation, while a similar treatment of the cultures of OLGcs that were isolated from 10-day-old animals did not induce further maturation of the cells. The results presented in the present report demonstrate that the in vivo effects of aTf on OLGc maturation can be reproduced in cultures and that the effects of aTf occur early in development during a narrow, transient "temporal window" within which OLGcs are sensitive to its action.

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Isolation of oligodendroglial cells from young and adult whole rat brains using an in situ generated percoll density gradient

Cited by 46 publications

References 19 publications

Sustained neonatal hyperthyroidism in the rat affects myelination in the central nervous system

Sustained neonatal hyperthyroidism in the rat affects myelination in the central nervous system

Thyroid Hormones and the Central Nervous System

Differential effects of apotransferrin on two populations of oligodendroglial cells

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