In situ hybridization localization of choline acetyltransferase mRNA in adult rat brain and spinal cord

Lauterborn, Julie C.; Isackson, Paul J.; Montalvo, Raúl; Gall, Christine M.

doi:10.1016/0169-328x(93)90073-x

Cited by 96 publications

(47 citation statements)

References 35 publications

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“…), or oleic acid (10 mg kg -1 ) and killed the rats 60 min later by decapitation under anaesthesia. In situ hybridization analyses were conducted using [ 35 S]-labelled antisense RNA probes for c-fos 28 and choline acetyl transferase 29 . Average hybridization densities were determined from at least three tissue sections per rat.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

An anorexic lipid mediator regulated by feeding

Fonseca

Navarro

Gómez

et al. 2001

Nature

645

767

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Oleylethanolamide (OEA) is a natural analogue of the endogenous cannabinoid anandamide. Like anandamide, OEA is produced in cells in a stimulus-dependent manner and is rapidly eliminated by enzymatic hydrolysis, suggesting a function in cellular signalling 1. However, OEA does not activate cannabinoid receptors and its biological functions are still unknown 2. Here we show that, in rats, food deprivation markedly reduces OEA biosynthesis in the small intestine. Administration of OEA causes a potent and persistent decrease in food intake and gain in body mass. This anorexic effect is behaviourally selective and is associated with the discrete activation of brain regions (the paraventricular hypothalamic nucleus and the nucleus of the solitary tract) involved in the control of satiety. OEA does not affect food intake when injected into the brain ventricles, and its anorexic actions are prevented when peripheral sensory fibres are removed by treatment with capsaicin. These results indicate that OEA is a lipid mediator involved in the peripheral regulation of feeding

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Section: In Situ Hybridizationmentioning

confidence: 99%

An anorexic lipid mediator regulated by feeding

Fonseca

Navarro

Gómez

et al. 2001

Nature

645

767

View full text Add to dashboard Cite

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“…In fact, small-medium cholinergic neurons and choline acetyltransferase mRNA have been identified in small cells of the NRGC/nucleus reticularis paragigantocellularis (NRPG). 7) These observations suggest that cholinergic neurons play an important role in the RVLM, including NRGC/NRGCa.…”

mentioning

confidence: 93%

Effects of Microinjected Carbachol on the Antinociceptive Response to Noxious Heat Stimuli.

Abe

Kikuta

Kato

et al. 2003

Biological & Pharmaceutical Bulletin

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The rostral ventrolateral medulla (RVLM), which includes the nucleus reticularis gigantocellularis (NRGC)/nucleus reticularis gigantocellularis alpha (NRGCa) and the lateral reticular nucleus, plays a major role in maintaining tonic vasomotor tone and is critical in the reflex control of blood pressure. 1,2) In addition, the RVLM modulates one of the physiological functions of antinociception via cholinergic neurons. 3,4) Immunohistochemical studies have shown that the RVLM is extensively innervated with cholinergic neurons 5) that mainly project from the pedunculopontine tegmental nucleus (PPT), 6) then efferently extend to the spinal cord. In fact, small-medium cholinergic neurons and choline acetyltransferase mRNA have been identified in small cells of the NRGC/nucleus reticularis paragigantocellularis (NRPG). 7) These observations suggest that cholinergic neurons play an important role in the RVLM, including NRGC/NRGCa.Anticholinesterase and cholinergic mimetics exert analgesic effects in humans, 8,9) monkeys, 10) rats and mice. 11)Moreover, the administration of carbachol into the nucleus raphe magnus induces pronounced antinociception that is reversed by atropine and muscarinic antagonists. 12) Injections of muscarinic agonists into the spinal cord also have antinociceptive effects on responses to noxious heat stimuli via muscarinic receptors. 11,13) We reported that either systemically-administered or RVLM-injected morphine enhances ACh release in the RVLM, and that the local application of morphine via a microdialysis probe induces an increase in both tail withdrawal and hot-plate responses.14) Thus, the cholinergic system in the central nervous system may be considered to have a key role in endogenous pain control.Thus, the present study investigates whether a direct injection of the cholinergic agonist, carbachol, into the NRGC/NRGCa evokes antinociception, and examines the participation of antinociceptive events in rats. MATERIALS AND METHODSAnimals Male Wistar rats, weighing 250 to 350 g at the time of the experiments, were purchased from Charles River Japan, Inc. All animals were housed individually under automatically controlled environmental conditions and 12 h light-dark cycles, and were handled in accordance with the guidelines for animal care and use published by the National Institute of Health. The rats had free access to food and water. All animals were quarantined in centralized animal facilities for at least seven days after arrival from the supplier.Preparation for Microinjection After an acclimation period, the animals were anesthetized with sodium pentobarbital (50 mg/kg, i.p.) and placed in a stereotaxic apparatus. A midline incision was made on the scalp, and a burr hole was drilled for a 27-gauge stainless-steel cannula (guide cannula) that was firmly placed using dental acrylic and anchored to the skull using stainless-steel screws. Drugs were injected through a 30-gauge stainless-steel cannula, the tip of which extended 4 mm beyond the tip of the guide cannula into the NRGC/NRGCa a...

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“…In some experiments, alternate sections were processed for in situ hybridization using ?S-labeled cRNA probes to localize mRNAs encoding either choline acetyltransferase (ChAT) (Lauterbom et al, 1993) or the p2 subunit of the nicotinic AChR (Deneris et al, 1988). Tissue sections were mounted on gelatin-coated slides following in situ hybridization, air dried, and exposed to B-Max hyperfilm (Amersham, Arlington Heights, IL) for 2-4 d. Mounted sections were subsequently defatted with chloroform, dipped in Kodak NTB2 emulsion (Eastman-Kodak, Rochester, NY), and exposed for 3-6 weeks at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Localization and alternative splicing of agrin mRNA in adult rat brain: transcripts encoding isoforms that aggregate acetylcholine receptors are not restricted to cholinergic regions

et al. 1994

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Agrin is a protein implicated in the formation and maintenance of the neuromuscular junction.In addition to motor neurons, agrin mRNA has been detected in the brains of embryonic rat and chick and adult marine ray, suggesting that this molecule may also be involved in the formation of synapses between neurons. As a step toward understanding agrin's role in the CNS, we utilized Northern blot and in situ hybridization techniques to analyze the regional distribution and cellular localization of agrin mRNA in the spinal cord and brain of adult rats. The results of these studies indicate that the agrin mRNA is expressed predominantly by neurons broadly distributed throughout the adult CNS. Moreover, expression of agrin mRNA is not restricted to cholinergic structures or regions of the brain receiving cholinergic input. Recently, RNA isolated from rat embryonic spinal cord was shown to contain four alternatively spliced agrin mRNAs, referred to as agrin,, agrin,, agrin,,, and agrin,,, each of which encodes agrin proteins that are active in acetylcholine receptor aggregating assays (Ferns et al., 1992). Using the polymerase chain reaction we demonstrate that all four of these agrin transcripts are expressed within the adult CNS. Agrin,, agrin,, and agrin,, were present in all regions analyzed. In contrast, agrin,, was detected only in forebrain. Results of these studies indicate that both the level of expression and pattern of alternative splicing of agrin mRNA are differentially regulated in the brain. The broad and predominantly neuronal distribution of agrin mRNA in the adult brain suggests that, in addition to its role at the neuromuscular junction, agrin may play a role in formation and maintenance of synapses between neurons in the CNS.

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In situ hybridization localization of choline acetyltransferase mRNA in adult rat brain and spinal cord

Cited by 96 publications

References 35 publications

An anorexic lipid mediator regulated by feeding

An anorexic lipid mediator regulated by feeding

Effects of Microinjected Carbachol on the Antinociceptive Response to Noxious Heat Stimuli.

Localization and alternative splicing of agrin mRNA in adult rat brain: transcripts encoding isoforms that aggregate acetylcholine receptors are not restricted to cholinergic regions

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