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Immuno-electron microscopy of sorting and release of neuropeptides in Lymnaea stagnalis

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Summary

The cerebral caudodorsal cells of the pulmonate snail Lymnaea stagnalis control egg laying and egglaying behavior by releasing various peptides derived from two precursors. The biosynthesis, storage, intracellular breakdown and release of three caudodorsal cell peptides were studied by means of immuno-electron microscopy using antisera raised to fragments of these peptides: (1) Caudodorsal Cell Hormone-I (CDCH-I; derived from precursor I), (2) Caudodorsal Cell Hormone-II (CDCH-II; from precursor II), and (3) α-Caudodorsal Cell Peptide (α CDCP; from both precursors). After affinity purification of the antisera, the specificity of the sera was confirmed with dotting immunobinding assays. From the ultrastructural immunocytochemical data it has been concluded that the precursor molecules are cleaved at the level of the Golgi apparatus after which the C-terminal parts (containing α CDCP) and N-terminal parts (containing CDCH-I or CDCH-II) are sorted and preferentially packaged into large electron-dense granules (MD 150 nm), respectively. Very probably, the content of the large electron-dense granules is degraded within the cell body. The immunoreactivity of the secretory granules increases during discharge from the Golgi apparatus, indicating further processing. At least a portion of the secretory granules contains all three peptides, as shown by double and triple immunopositive stainings whereas other granules appear to contain only one or two of these peptides. The caudodorsal cells release multiple peptides via exocytosis from neurohemal axon terminals into the hemolymph and from blindly ending axon collaterals into the intercellular space of the cerebral commissure (nonsynaptic release).

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Author notes

  1. E. W. Roubos

    Present address: Department of Animal Physiology, University of Nijmegen, NL-6525 ED, Nijmegen, The Netherlands

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  1. Department of Biology, Vrije Universiteit, P.O. Box 7161, NL-1007 MC, Amsterdam, The Netherlands

    W. R. A. van Heumen & E. W. Roubos

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  1. W. R. A. van Heumen
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van Heumen, W.R.A., Roubos, E.W. Immuno-electron microscopy of sorting and release of neuropeptides in Lymnaea stagnalis . Cell Tissue Res 264, 185–195 (1991). https://doi.org/10.1007/BF00305737

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