CHARACTERIZATION OF AN α‐BUNGAROTOXIN BINDING COMPONENT FROM <i>DROSOPHILA MELANOGASTER</i>

Schmidt‐Nielsen, Bodil; Gepner, Janice; Teng, Nelson N.H.; Hall, Linda M.

doi:10.1111/j.1471-4159.1977.tb06505.x

Cited by 150 publications

(54 citation statements)

References 31 publications

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“…Additionally, ACh receptors have been localized in Drosophila medulla and lobula with radiolabeled alpha-bungarotoxin (Schmidt-Nielsen et al, 1977). The putative receptors exhibit a tangentially stratified distribution in the medulla and anterior and posterior lobula.…”

Section: Discussionmentioning

confidence: 99%

“…The above features make the optic lobe an attractive subject for the cytochemical analysis of neurotransmitters. The earliest studies used a variety of methods such as formaldehyde-induced fluorescence for catecholamines (Elofsson and Klemm, 1972;Elofsson et al, 1977), alpha-bungarotoxin binding for ACh receptors (Schmidt-Nielsen et al, 1977;Maxwell and Hildebrand, 198 1) and the incorporation of radioactive precursors (CamposOrtega, 1974;Maxwell and Hildebrand, 198 1;Elias and Evans, 1984). Quantitative assays have been based upon neurotransmitter extraction from optic lobe fragments and analysis by electrophoresis and high-performance liquid chromatography (Maxwell et al, 1978;Maxwell and Hildebrand, 198 1;Elofsson et al, 1982;Elias and Evans, 1983;Garcia and Arechiga, 1986).…”

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

et al. 1989

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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.

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Section: Discussionmentioning

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

et al. 1989

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“…Published procedures for binding iodinated toxin to lightly fixed frozen sections (19,20) were used for Manduca brain. The brain was bathed in cold paraformaldehyde fixative solution (4% wt/vol paraformaldehyde containing 0.14 M sucrose and 0.075 M sodium phosphate buffer, pH 7.4) during dissection.…”

Section: Introductionmentioning

confidence: 99%

“…compound. The sectioning, incubation with radioactive toxin, and subsequent processing were done as described (20). Toxin (90-154 Ci/ mmol; 1 Ci = 3.7 X 1010 becquerels) was used in the concentration range of 25-45 nM, and autoradiograms were developed after exposures of 6-20 days.…”

Section: Introductionmentioning

confidence: 99%

“…To probe for AcCho receptors in the antennal lobes, we have used the snake venom protein a-bungarotoxin (a-Bgt), which is a powerful agent for identifying and quantifying nicotinic AcCho receptors at vertebrate neuromuscular junctions (17,18). Previous studies have shown that the insect central nervous system contains an a-Bgt-binding component that exhibits several properties expected of AcCho receptors (19)(20)(21)(22)(23). A similar a-Bgt-binding activity is present in detergent-solubilized extracts of the antennal lobes of Manduca (16).…”

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Distribution of binding sites for 125I-labeled alpha-bungarotoxin in normal and deafferented antennal lobes of Manduca sexta.

Hildebrand¹,

Hall²,

Osmond³

1979

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

118

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ABSTRACT'25I-Labeled a-bungarotoxin has been used to determine the distribution of putative acetylcholine receptors in normal and chronically deafferented antennal lobes in the brain of the moth Manduca sexta. Toxin-binding sites are confined to synaptic regions in the neuropil of normal lobes and to rudiments of these regions in deafferented lobes. These findings suggest that receptors can develop in the insect central nervous system independently of normal synaptic influences. Many excitable cells require innervation for their normal development and for maintenance of their normally differentiated properties (for examples, see reviews 1-3). The most fully documented case of this role of innervation is the regulation of the distribution of acetylcholine (AcCho) receptors in vertebrate skeletal muscle fibers developing in vivo (4-7) and in vitro (8-13). AcCho receptors, which appear about the time of myotube formation, are initially distributed over the entire surface of the myotubes. As the developing muscle fiber becomes innervated and synaptic transmission begins, AcCho receptors are lost from the extrajunctional regions of the muscle fiber but are retained in the junctional membrane. If innervation is prevented, this redistribution of receptors does not occur, and also the immature muscle eventually dies. From these observations it appears that the motor innervation influences the muscle fiber to localize its AcCho receptors in the subsynaptic membrane.We know of no comparable study of development of receptors during de novo synapse formation in the central nervous system. Therefore, we have used the antennal lobes of the brain of the moth Manduca sexta to study the role of normal innervation in regulating the development and distribution of AcCho receptors on central neurons. The antennal lobes are an excellent preparation for such studies because they appear to have a high density of cholinergic synapses, a simple anatomical organization with clear separation of synaptic and nonsynaptic regions, and experimentally observable and manipulable development (14-16). To probe for AcCho receptors in the antennal lobes, we have used the snake venom protein a-bungarotoxin (a-Bgt), which is a powerful agent for identifying and quantifying nicotinic AcCho receptors at vertebrate neuromuscular junctions (17,18 Deafferentation involved amputation of antennal rudiments ("deantennation") on day 1 after the larval-pupal molt (16). This operation removes the epidermis in which the antennal sensory neurons ordinarily arise and is performed prior to the birthdays of these neurons.Histology. For dissection of the brain and subesophageal ganglion, a developing or pharate adult Manduca was anesthetized with CO2, and the brain was exposed through a window cut in the head cuticle (16). The brain was moistened with cold physiological salts solution during dissection (16). Tissues were fixed in modified Serra's fixative (25), dehydrated, and embedded in Paraplast Plus (Sherwood Medical Industries, St. Louis, MO). They were sectione...

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Binding of Acetylcholine Receptor/Channel Probes to Housefly Head Membranes

Eldefrawi

Shaker

Eldefrawi

1982

Ciba Foundation Symposium 88 ‐ Neuropharmacology of Insects

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CHARACTERIZATION OF AN α‐BUNGAROTOXIN BINDING COMPONENT FROM DROSOPHILA MELANOGASTER

Cited by 150 publications

References 31 publications

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

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

Distribution of binding sites for 125I-labeled alpha-bungarotoxin in normal and deafferented antennal lobes of Manduca sexta.

Binding of Acetylcholine Receptor/Channel Probes to Housefly Head Membranes

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