Cellular Development and Myelin Production in Primary Cultures of Embryonic Mouse Brain

Fabre, M.; Langley, O.K.; Bologa, Liane; Delaunoy, Jean‐Pierre; Löwenthal, A.; Ferret-Sena, Véronique; Vincendon, G.; Sarliève, Louis L.

doi:10.1159/000112300

Cited by 19 publications

(24 citation statements)

References 18 publications

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“…Using other homogenates, the measurements of MK-4-d 7 and MK-4 epoxide-d 7 were carried out with the same LC-APCI-MS/MS method as described above. Primary cultures of mouse cerebral hemispheres on embryonic day (E) 14 were prepared as described previously with minor modifications (26). Fetuses (E14) were removed from pregnant C57BL/6J mice.…”

Section: Measurements Of Pk Mk-4 and Their Respective Epoxides In Tmentioning

confidence: 99%

Conversion of Phylloquinone (Vitamin K1) into Menaquinone-4 (Vitamin K2) in Mice

Okano

Shimomura

Yamane

et al. 2008

Journal of Biological Chemistry

229

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There are two forms of naturally occurring vitamin K, phylloquinone and the menaquinones. Phylloquinone (vitamin K 1 ) is a major type (>90%) of dietary vitamin K, but its concentrations in animal tissues are remarkably low compared with those of the menaquinones, especially menaquinone-4 (vitamin K 2 ), the major form (>90%) of vitamin K in tissues. Despite this great difference, the origin of tissue menaquinone-4 has yet to be exclusively defined. It is postulated that phylloquinone is converted into menaquinone-4 and accumulates in extrahepatic tissues. To clarify this, phylloquinone with a deuterium-labeled 2-methyl-1,4-naphthoquinone ring was given orally to mice, and cerebra were collected for D NMR and liquid chromatography-tandem mass spectrometry analyses. We identified the labeled menaquinone-4 that was converted from the given phylloquinone, and this conversion occurred following an oral or enteral administration, but not parenteral or intracerebroventricular administration. By the oral route, the phylloquinone with the deuterium-labeled side chain in addition to the labeled 2-methyl-1,4-naphthoquinone was clearly converted into a labeled menaquinone-4 with a non-deuterium-labeled side chain, implying that phylloquinone was converted into menaquinone-4 via integral side-chain removal. The conversion also occurred in cerebral slice cultures and primary cultures. Deuterium-labeled menadione was consistently converted into the labeled menaquinone-4 with all of the administration routes and the culture conditions tested. Our results suggest that cerebral menaquinone-4 originates from phylloquinone intake and that there are two routes of accumulation, one is the release of menadione from phylloquinone in the intestine followed by the prenylation of menadione into menaquinone-4 in tissues, and another is cleavage and prenylation within the cerebrum.

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Section: Measurements Of Pk Mk-4 and Their Respective Epoxides In Tmentioning

confidence: 99%

Conversion of Phylloquinone (Vitamin K1) into Menaquinone-4 (Vitamin K2) in Mice

Okano

Shimomura

Yamane

et al. 2008

Journal of Biological Chemistry

229

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“…1). It has been previously shown in our labora tory that this phase could be related to neu ronal multiplication and possibly also to the beginning of glial cell multiplication [36,37], From 15 to 20 DIV the DNA level increased again by about 30%, which could be due to glial cell multiplication, remained steady up to 25 DIV and subsequently declined slowly ( fig. 1).…”

Section: Dna and Protein Content And Activities O F Cst Cnp Asa Andmentioning

confidence: 53%

“…2c) as a function of time in culture. For CST, two peaks of activity were observed: a smaller one when the culture system is neuron-en riched (7 DIV) and a higher one at a time (21-25 days) corresponding to oligodendroglial differentiation [37,38]. In this cul ture system, CNP activity ( fig.…”

Section: Dna and Protein Content And Activities O F Cst Cnp Asa Andmentioning

confidence: 84%

“…In an attempt to clarify the role of this hormone on brain maturation, we have tested the influence of low concentrations (2 and 20 ng/ml) of insulin on two myelin-re lated enzymes: (1) the 3'-phosphoadenosine-5'-phosphosulfate:cerebroside sulfotransferase (PAPS:CST or CST, EC 2.8.2.11) catalyzing the production of sulfogalactosylceramide (sulfatide), one of the major my elin glycolipids [30] which is considered to be associated with CNS myelination [31,32], and (2) the myelin enzyme 2',3'-cyclic nuc leotide 3'-phosphohydrolase (CNP, EC 3.1.4.3.7) known to be a good marker of oligodendrocytes [33]. In parallel, we have tested the effect of insulin on arylsulfatase A (ASA, EC 3.1.6.1) and on cerebroside sulfatase (CS), both of which are involved in sul fatide degradation [34,35], To test the effect of insulin on the activity of these enzymes we have used a monolayer primary culture system of cells dissociated from brain hemispheres of 14-day-old mouse embryos, which is neuron-enriched from about 1 to 12 days in vitro (DIV) and glialenriched between about 12 and 60 days DIV [36,37]. Furthermore, myelin-related membranes are produced in this culture system [37,38].…”

Section: Introductionmentioning

confidence: 99%

“…In parallel, we have tested the effect of insulin on arylsulfatase A (ASA, EC 3.1.6.1) and on cerebroside sulfatase (CS), both of which are involved in sul fatide degradation [34,35], To test the effect of insulin on the activity of these enzymes we have used a monolayer primary culture system of cells dissociated from brain hemispheres of 14-day-old mouse embryos, which is neuron-enriched from about 1 to 12 days in vitro (DIV) and glialenriched between about 12 and 60 days DIV [36,37]. Furthermore, myelin-related membranes are produced in this culture system [37,38]. As previously shown, this system was also found to be very useful for studying the regulation of myelination by thyroid hormones.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of the Mechanisms of Action of Insulin and Triiodothyronine on the Synthesis of Cerebroside Sulfotransferase in Cultures of Cells Dissociated from Brains of Embryonic Mice

Ferret-Sena¹,

Sena²,

Besnard³

et al. 1990

Dev Neurosci

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The effect of low (physiological) concentrations of insulin (2 and 20 ng/ml) and L -triiodothyronine (T₃) were studied on two myelin-related enzymes: (1) the 3''-phospho-adenosine-5''-phosphosulfate:cerebroside sulfotransferase (CST, EC 2.8.2.11) catalyzing the production of sulfatide, and (2) the myelin enzyme, 2'',3''-cyclic nucleotide 3''-phosphohydrolase (CNP, EC 3.1.4.3.7) in myelinogenic cultures of cells dissociated from embryonic mouse brain. Insulin treatment (20 ng/ml) of the cells in the presence of serum increased CST activity at 18 and 25 days in vitro (DIV) by 86 and 211%, respectively. At 18 DIV and under the same conditions, CNP was significantly stimulated (95%) by high doses of insulin (2,000 ng/ml) only, while arylsulfatase A (EC 3.1.6.1) or cerebroside sulfatase activities, both of which are involved in sulfatide degradation, were unchanged. Thus, it can be assumed that the observed increase of the incorporation of [³⁵S]0₄ into sulfatide after insulin treatment of mixed cell cultures is the result of CST induction rather than a decreased catabolism. The level of CST activity in insulin-treated cells (20 ng/ml) in serum-free medium was also increased at 18 and 25 DIV by about 50 and 70%, respectively. Conversely, none of the insulin concentrations used in the absence of serum (even at high doses) had any effect, either at 18 or 25 DIV on CNP and ASA activities. The involvement of insulin in the regulation of sulfatide synthesis was further confirmed by dose-response curves relating the activity of CST to hormone concentration in the medium. The increase in the activity of CST in insulin-treated cells was due only to the increase in the V_max of this enzyme, suggesting that it may be attributed to enzyme induction. A study of kinetic parameters of CST indicated that there were no differences in pH optimum and K_m values between control and induced enzyme. Further experiments using cycloheximide point to a direct effect of insulin on oligodendrocyte CST induction. Data similar to those described above for insulin were also obtained with T₃. As for insulin, T₃ stimulated the induction of CST but in serum-free medium only. This effect was prevented by cycloheximide. In addition, the induction of CST by T₃ was blocked by actinomycin D. This was not the case for insulin. These results suggest that T₃ and insulin act on CST by different mechanisms, i.e. at transcriptional and post-translational levels, respectively. Apart from this, the insulin effect on CST activity was additive to that of T₃. Finally, in secondary cultures of pure rat oligodendrocytes, which are capable of producing structures resembling compact myelin, high doses of insulin (5 µg/ml) increased the incorporation of 5-[¹²⁵I]iodo-2''-deoxyuridine by 68%. In contrast, T₃ did not stimulate oligodendrocyte proliferation. It can be concluded that both hormones may have specific effects on myelination.

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Cultures of cells from fetal rat brain: Methods to control composition, morphology, and biochemical activity

Mahoney

Saltzman

1999

Biotechnol. Bioeng.

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Cellular Development and Myelin Production in Primary Cultures of Embryonic Mouse Brain

Cited by 19 publications

References 18 publications

Conversion of Phylloquinone (Vitamin K1) into Menaquinone-4 (Vitamin K2) in Mice

Conversion of Phylloquinone (Vitamin K1) into Menaquinone-4 (Vitamin K2) in Mice

Comparison of the Mechanisms of Action of Insulin and Triiodothyronine on the Synthesis of Cerebroside Sulfotransferase in Cultures of Cells Dissociated from Brains of Embryonic Mice

Cultures of cells from fetal rat brain: Methods to control composition, morphology, and biochemical activity

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