Immunocytochemical Detection of Acetylcholine in the Rat Central Nervous System

Geffard, M.; McRae-Degueurce, A.; Souan, Marie-Laure

doi:10.1126/science.3892687

Cited by 71 publications

(23 citation statements)

References 22 publications

(5 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Initial studies compared SMI-32 labeling of motor neurons in slices with that of various other markers reported to stain motor neurons. Antibodies against the following markers were examined: the neurotransmitter ACh (Geffard et al, 1985), the ACh-synthesizing enzyme ChAT (Hietanen et al, 1990;Wang et al, 1990), peripherin (Parysek and Goldman, 1988;Escurat et al, 1990), a class III intermediate filament protein, and CGRP (Gibson et al, 1984;Kruger et al, 1988;Juurlink et al, 1990). Although each of these markers labeled motor neurons in the ventral horn, the degree of staining varied significantly (Fig.…”

Section: Smi-32 and Other Motor Neuron Markers Label Motor Neurons Inmentioning

confidence: 99%

Motor Neurons Are Selectively Vulnerable to AMPA/Kainate Receptor-Mediated InjuryIn Vitro

1996

View full text Add to dashboard Cite

The nonphosphorylated neurofilament marker SMI-32 stains motor neurons in spinal cord slices and stains a subset of cultured spinal neurons neurons"], which have a morphology consistent with motor neurons identified in vitro: large cell body, long axon, and extensive dendritic arborization. They are found preferentially in ventral spinal cord cultures, providing further evidence that large SMI-32(ϩ) neurons are indeed motor neurons, and SMI-32 staining often colocalizes with established motor neuron markers (including acetylcholine, calcitonin gene-related peptide, and peripherin). Additionally, choline acetyltransferase activity (a frequently used index of the motor neuron population) and peripherin(ϩ) neurons share with large SMI-32(ϩ) neurons an unusual vulnerability to AMPA /kainate receptor-mediated injury. Kainateinduced loss of these motor neuron markers is Ca 2ϩ -dependent, which supports a critical role of Ca 2ϩ ions in this injury. Raising extracellular Ca 2ϩ exacerbates injury, whereas removal of extracellular Ca 2ϩ is protective. A basis for this vulnerability is provided by the observation that most peripherin(ϩ) neurons, like large SMI-32(ϩ) neurons, are subject to kainate-stimulated Co 2ϩ uptake, a histochemical stain that identifies neurons possessing Ca 2ϩ -permeable AMPA /kainate receptor-gated channels. Finally, of possibly greater relevance to the slow motor neuronal degeneration in diseases, both large SMI-32(ϩ) neurons and peripherin(ϩ) neurons are selectively damaged by prolonged (24 hr) low-level exposures to kainate (10 M) or to the glutamate reuptake blocker L-transpyrrolidine-2,4-dicarboxylic acid (100 M). During these lowlevel kainate exposures, large SMI-32(ϩ) neurons showed higher intracellular Ca 2ϩ concentrations than most spinal neurons, suggesting that Ca 2ϩ ions are also important in this more slowly evolving injury.

show abstract

Section: Smi-32 and Other Motor Neuron Markers Label Motor Neurons Inmentioning

confidence: 99%

Motor Neurons Are Selectively Vulnerable to AMPA/Kainate Receptor-Mediated InjuryIn Vitro

1996

View full text Add to dashboard Cite

show abstract

“…In the present study, ACh and choline were assayed in the crayfish optic lobe, and the highest concentrations of both substances were found in the medulla externa and the medulla interna (second and third optic neuromeres). Additionally, we have examined the optic lobe with antisera developed for the immunocytochemical detection of ACh (Geffard et al, 1985). We found ACh-like immunoreactivity in a centrifugal neuron of the lamina and a subset of the columnar visual intemeurons (transmedullary neurons) of the second and third optic neuromeres.…”

mentioning

confidence: 98%

Acetylcholine in the crayfish optic lobe: concentration profile and cellular localization

et al. 1989

View full text Add to dashboard Cite

The crayfish optic lobe contains high levels of acetylcholine (ACh) and choline as measured with a chemiluminescent assay in small fragments of optic lobe tissue. The highest concentrations were found in the medulla externa and medulla interna (second and third optic neuromeres), which have ACh concentrations of 270 pmol/mg tissue. This concentration is about 16 times that measured in the photoreceptors and lamina ganglionaris (the first optic neuromere). Immunocytochemistry (based upon antisera to choline-glutaryl-BSA) revealed low levels of ACh-like reactivity in the lamina ganglionaris associated with the terminal arbors of centrifugal and/or tangential neurons. The most intense ACh- like reactivity was observed in monopolar neurons of the medulla externa and medulla interna. One monopolar neuron/medullary column (or about 2500 neurons/medullary neuropile) exhibited reactivity and an estimated cytoplasmic concentration of 8.1 mM.

show abstract

“…(iv) The immunocytochemical reaction was observed only after perfusion with a mixture previously demonstrated to fix AcCho in the rat CNS (9,10 Nor could staining be observed in 'the dopaminergic cells of the substantia nigra region of glutaraldehyde/allyl alcoholfixed rat brain after incubation with the CSF from any AD patient.…”

Section: Resultsmentioning

confidence: 94%

“…To achieve this, rats were perfused with a chemical mixture previously shown to fix small neurotransmitter molecules such as AcCho in the CNS (9,10). Light microscopic examinations revealed distinct immunoreactivity in neurons in the MS and in the VDB after incubations with rabbit anti-AcCho antiserum or with the CSF from six of the eight AD patients.…”

Section: Resultsmentioning

confidence: 99%

“…The specificity of the AcCho antiserum (A31) has been investigated immunocytochemically by the group of Geffard, using an approach previously described (9). AcChoEase staining has been shown previously to identify cholinergic [choline acetyltransferase (ChoATase) immunoreactive] neurons in the rat brain after short-term treatment with irreversible AcChoEase inhibitors (14).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Antibodies in cerebrospinal fluid of some Alzheimer disease patients recognize cholinergic neurons in the rat central nervous system.

McRae-Degueurce

Bööj

Haglid

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The etiology of Alzheimer disease is unclear. However, immunological aberrations have been suggested to be critical factors in the pathogenesis of this neurodegenerative disease. This study was carried out to investigate if cerebrospinal fluid (CSF) from Alzheimer disease patients contains antibodies that recognize specific neuronal populations in the rat central nervous system. The results indicate that in a subgroup of patients this is indeed the case. The antibodies reported in this study have the following properties: (i) they recognize neuronal populations and components in the medial septum and spinal motor neurons in rats perfused with a mixture that fixes small neurotransmitter molecules; (ii) adsorption of the patient CSF with staphylococcal protein ASepharose and using a polyclonal antiserum against human IgG3 indicates that the immunocytochemical reaction in these brain regions is mainly due to the subclass IgG3; and (iii) the CSF immunocytochemical reaction is blocked by preincubation of the sections with a rabbit anti-acetylcholine antiserum. These results provide evidence that antibodies in the CSF of some, but not all, Alzheimer disease patients recognize acetylcholine-like epitopes in cholinergic neurons in the rat central nervous system. Alzheimer disease (AD) is a neurodegenerative disorder characterized by loss of memory, perception, reasoning, orientation, and judgement. The impairment of intellectual function has been correlated with abnormal morphological phenomena, the most prominent being neurofibrillary tangles and neuritic plaques as well as the loss of cholinergic presynaptic markers (1)(2)(3)(4) (8). Clinical examination confirmed dementia of mild to moderate degree. The patients had no symptoms corresponding to other neurological disorders. The CSF samples (12 ml) were collected on ice, gently mixed to avoid gradient effects, centrifuged to remove cells, divided into 2-ml aliquots, and immediately frozen at -20'C without preservatives. At the time of sampling the patients were not receiving pharmacological treatment. A more detailed description of patient history will be given elsewhere.Immunocytochemical Approach. Three male rats (SpragueDawley) were perfused with a mixture of 5% (wt/vol) glutaraldehyde and allyl alcohol (1 M) in a sodium cacodylate buffer (0.5 M, pH 12) (9, 10). After thorough washing in Tris base (0.5 M) containing 1% sodium metabisulfite (adjusted to pH 7.6 with concentrated sodium hydroxide) the brains were rinsed overnight in the same buffer containing 10% sucrose and then frozen with compressed CO2 gas. Brains were sectioned (10 ,um) in a cryostat. Serial frozen sections of the medial septum (MS) and vertical limb of the diagonal band (VDB), or of the spinal cord, were mounted on gelatin-coated glass slides and incubated with 2% normal goat serum for 30 min. The serial sections of the MS or spinal cord were incubated overnight either with rabbit antiserum to acetylcholine (AcCho) (9) (1:500 or 1:200) or with the CSF from the eight AD patients (1:1) dil...

show abstract

Immunocytochemical Detection of Acetylcholine in the Rat Central Nervous System

Cited by 71 publications

References 22 publications

Motor Neurons Are Selectively Vulnerable to AMPA/Kainate Receptor-Mediated InjuryIn Vitro

Motor Neurons Are Selectively Vulnerable to AMPA/Kainate Receptor-Mediated InjuryIn Vitro

Acetylcholine in the crayfish optic lobe: concentration profile and cellular localization

Antibodies in cerebrospinal fluid of some Alzheimer disease patients recognize cholinergic neurons in the rat central nervous system.

Contact Info

Product

Resources

About