Regulation of neuronal differentiation in neuron‐enriched primary cultures from embryonic rat cerebra by platelet activating factor and the structurally related glycerol ether lipid, dodecylglycerol

Ved, H. S.; Gustow, Evan; Pieringer, Ronald A.

doi:10.1002/jnr.490300211

Cited by 26 publications

(11 citation statements)

References 22 publications

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“…AChE levels were determined by the Ellman's method that was adapted for 96-well plates [Ved et al, 1991;Dave et al, 1997]. For the AChE assay, an increase in absorbance was monitored at 412 nm for 10 min in a reaction mixture containing 20 ml extracellular or intracellular enzyme as described below, 10 ml of 30 mM acetylthiocholine iodide (ATC), 10 mL of 10 mM dithionitrobenzene, and 50 mM sodium phosphate buffer (pH 8) in a final volume of 300 mL.…”

Section: Ache Microassaymentioning

confidence: 99%

Histone acetylase inhibitor trichostatin A induces acetylcholinesterase expression and protects against organophosphate exposure

Curtin

Tetz

Compton

et al. 2005

J of Cellular Biochemistry

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The biological effects of organophosphorous (OP) chemical warfare nerve agents (CWNAs) are exerted by inhibition of acetylcholinesterase (AChE), which prevents the hydrolysis of the neurotransmitter acetylcholine, leading to hypercholinergy, seizures/status epilepticus, respiratory/cardiovascular failure, and potentially death. Current investigations show that bioscavenger therapy using purified fetal bovine AChE in rodents and non-human primates and the more recently tested human butyrylcholinesterase, is a promising treatment for protection against multiple LD(50) CWNA exposures. Potential impediments, due to the complex structure of the enzyme, purification effort, resources, and cost have necessitated alternative approaches. Therefore, we investigated the effects of transcriptional inducers to enhance the expression of AChE to achieve sufficient protection against OP poisoning. Trichostatin A (TSA), an inhibitor of histone deacetylase that de-condenses the chromatin, thereby increasing the binding of transcription factors and mRNA synthesis, was evaluated for induction of AChE expression in various neuronal cell lines. Dose-response curves showed that a concentration of 333 nM TSA was optimal in inducing AChE expression. In Neuro-2A cells, TSA at 333 nM increased the extracellular AChE activity approximately 3-4 fold and intracellular enzyme activity 10-fold. Correlating with the AChE induction, TSA pre-treatment significantly protected the cells against exposure to the organophosphate diisopropylfluorophosphate, a surrogate for the chemical warfare agents soman and sarin. These studies indicate that transcriptional inducers such as TSA up-regulate AChE, which then can bioscavenge any organophosphates present, thereby protecting the cells from OP-induced cytotoxicity. In conclusion, transcriptional inducers are prospective new methods to protect against CWNA exposure.

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Section: Ache Microassaymentioning

confidence: 99%

Histone acetylase inhibitor trichostatin A induces acetylcholinesterase expression and protects against organophosphate exposure

Curtin

Tetz

Compton

et al. 2005

J of Cellular Biochemistry

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“…The pellet was suspended in 500 pi PBS and stored at -7 0°C until analyzed. Enrichment of neuronal culture was assessed by cell-type specific neurochemical, immunological techniques and was found to be 90-95% enriched in neurons [15]. For this purpose, acetylcholinesterase (AChE) as well as choline acetyltransferase (ChAT; neuronal markers), myelin basic pro tein and cyclic nucleotide phosphohydrolase (oligodendroglial markers) and glutamine synthetase (astroglial marker) were mea sured [15].…”

Section: Materials and M Ethodsmentioning

confidence: 99%

“…Enriched neuronal cultures were prepared from 17-day-old rat embryos as previously described [15]. Embryos were removed by cesarean section and were placed in a Petri dish containing defined medium (DMEM/F12) supplemented with 20% equine serum (HyClone Lab, Logan, Utah, USA) [16], Cerebra were removed by gross dissection and as many meninges were removed as possible.…”

Section: Materials and M Ethodsmentioning

confidence: 99%

“…A unit was de fined as a micromole of ATCI hydrolyzed per minute. Although the neuronal specificity of AChE has been questioned in some systems [18], it is a known good indicator of neuronal content and condition in this culture system [15], ChAT was assayed as described by Fonnum [19]. Twenty mi EDTA [pH 7.4, 0.1 mA/ physostigmine) and 8 pi cell homogenate was incubated for 30 min at 37°C.…”

Section: Materials and M Ethodsmentioning

confidence: 99%

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Regulation of Neuronal Differentiation by Retinoic Acid Alone and in Cooperation with Thyroid Hormone or Hydrocortisone

Ved

Pieringer

1993

Dev Neurosci

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Cultures highly enriched in neurons obtained from embryonic mouse cerebra were used to demonstrate that: (1) at the optimum concentration of 10^–8M retinoic acid stimulated the neurons to produce axon- and dentrite-like structures as determined by phase contrast and fluorescent microscopy; (2) the same concentration of retinoic acid stimulated acetyl cholinesterase and choline acetyltransferase activities; (3) treatment of neurons of either prenatal or neonatal equivalent age with retinoic acid produced a sustained stimulation of neuronal differentiation, and (4) retinoic acid cooperatively stimulated neuronal differentiation with either thyroid hormone or hydrocortisone.

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“…On day 4 the medium and any unattached cells were replaced by a serum-free chemically defined medium. Cultures were enriched (90-95% ) in neurons by treatment with 9.87 pM cytosine arabinoside [19]. The test groups contained either HC (0.3 |iM), T3 (20 nM), RA (10 7 M), Tj + RA.…”

Section: Preparation O F Cerebral Mixed and Neuronal Cell Culturesmentioning

confidence: 99%

Regulation of Neuronal Thyroid Hormone Receptor α<sub>1</sub> mRNA by Hydrocortisone, Thyroid Hormone and Retinoic Acid

Satyanarayana¹,

Sarvesh²,

Khadeer³

et al. 1994

Dev Neurosci

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The regulation of thyroid hormone receptor Α₁(TRΑ₁) mRNA by hydrocortisone (HC), thyroid hormone (T₃) and retinoic acid (RA) has been studied in mixed and neuronal primary cultures of cells dissociated from fetal rat cerebra. Steady-state levels of TRΑ₁ mRNA were increased as much as 5-fold at 13 days of development by 0.3 µM HC in both mixed and neuronal-enriched cultures. The regulation of TRΑ₁ mRNA by HC appears to be mainly limited to neurons. This conclusion is based on two observations. First, stimulation by HC occurs in cultures highly enriched in neurons at approximately the same time and extent as that seen in mixed-cell cultures (containing neurons, oligodendroglia and astroglia). Second, the stimulation reaches a peak at 13 days in mixed-cell cultures, an age at which neurons but not glia differentiate.

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Regulation of neuronal differentiation in neuron‐enriched primary cultures from embryonic rat cerebra by platelet activating factor and the structurally related glycerol ether lipid, dodecylglycerol

Cited by 26 publications

References 22 publications

Histone acetylase inhibitor trichostatin A induces acetylcholinesterase expression and protects against organophosphate exposure

Histone acetylase inhibitor trichostatin A induces acetylcholinesterase expression and protects against organophosphate exposure

Regulation of Neuronal Differentiation by Retinoic Acid Alone and in Cooperation with Thyroid Hormone or Hydrocortisone

Regulation of Neuronal Thyroid Hormone Receptor α<sub>1</sub> mRNA by Hydrocortisone, Thyroid Hormone and Retinoic Acid

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