Abstract
Immunocytochemistry with monoclonal antibodies was used to investigate the locations of muscarinic acetylcholine receptors (mAChR) and choline acetyltransferase (ChAT) in sections of the developing antennae of the moth Manduca sexta. The results were correlated with a previous morphological investigation in the developing antennae which allowed us to locate different cell types at various stages of development. Our findings indicated that the muscarinic cholinergic system was not restricted to the sensory neurons but was also present in glial and epidermal cells. By day 4–5 of adult development, immunoreactivity against both antibodies was present in the axons of the antennal nerve, and more intense labeling was present in sections from older pupae. At days 4–9, the cell bodies of the sensory neurons in the basal part of the epidermis were also intensely immunolabeled by the anti-mAChR antibody. In mature flagella, large numbers of cells, some with processes into hairs, were strongly labeled by both antibodies. Antennal glial cells were intensely immunolabeled with both antibodies by days 4–5, but in later stages, it was not possible to discriminate between glial and neural staining. At days 4–9, we observed a distinctly labeled layer of epidermal cells close to the developing cuticle. The expression of both ChAT and mAChRs by neurons in moth antennae may allow the regulation of excitability by endogenous ACh. Cholinergic communication between neurons and glia may be part of the system that guides axon elongation during development. The cholinergic system in the apical part of the developing epidermis could be involved in cuticle formation.
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Acknowledgements
We thank Andrew S. French for critical reading of the manuscript. The original monoclonal antibody against ChAT (4B1) was a donation from Dr. Paul Salvaterra (Beckman Research Institute of the City of Hope, Calif.) and the monoclonal antibody against MPA was a gift from Drs. John G. Hildebrand and Lynne A. Oland (Arizona Research Laboratories, Tucson, Ariz.). We are grateful to Monika Stengl, Harjit Seyan, and Ian A. Meinertzhagen for advice regarding the initiation of the histological procedures.
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This work was supported by grants from the Natural Sciences and Engineering Research Council of Canada (NSERC), the Canadian Foundation for Innovation, and the Nova Scotia Research and Innovation Trust to P.H.T. and a NSERC postdoctoral fellowship to J.C.
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Clark, J., Meisner, S. & Torkkeli, P.H. Immunocytochemical localization of choline acetyltransferase and muscarinic ACh receptors in the antenna during development of the sphinx moth Manduca sexta. Cell Tissue Res 320, 163–173 (2005). https://doi.org/10.1007/s00441-004-1039-7
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DOI: https://doi.org/10.1007/s00441-004-1039-7