Input and output synapses on identified motor neurones of a locust revealed by the intracellular injection of horseradish peroxidase

Summary

Physiologically characterised motor neurones in the thoracic ganglia of the locust were injected with horseradish peroxidase in order that the spatial relationship between their input and output synapses could be observed with the electron microscope. A modification in the development procedure for the peroxidase ensured that the internal fine structure of the stained neurones was not obscured by the diaminobenzidine reaction product. Input and output synapses may occur within 1 μm of each other on the neuropilar processes of the motor neurones. This supports physiological evidence that motor neurones may be involved in local circuit interactions within the thoracic ganglia.

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References

  1. Adams JC (1977) Technical considerations in the use of horseradish peroxidase as a neuronal marker. Neuroscience 2:141–145

    Google Scholar 

  2. Altman JS, Shaw MK, Tyrer NM (1979) Visualisation of synapses of physiologically identified cobaltfilled neurones in the locust. J Physiol 296:2P.

    Google Scholar 

  3. Altman JS, Shaw MK, Tyrer NM (1980) Input synapses onto a locust sensory neurone revealed by cobalt-electron microscopy. Brain Res 189:245–250

    Google Scholar 

  4. Burrows M (1973) The role of delayed excitation in the coordination of some metathoracic flight motoneurones of a locust. J Comp Physiol 83:135–164

    Google Scholar 

  5. Burrows M (1975) Monosynaptic connexions between wing stretch receptors and flight motoneurones of the locust. J Exp Biol 62:189–219

    Google Scholar 

  6. Burrows M (1979) Synaptic potentials effect release of transmitter from locust nonspiking interneurones. Science 204:81–83

    Google Scholar 

  7. Burrows M, Siegler MVS (1978) Graded synaptic transmission between local interneurones and motor neurones in the metathoracic ganglion of the locust. J Physiol 285:231–255

    Google Scholar 

  8. Christensen BN (1973) Procion brown, an intracellular dye for light and electron microscopy. Science 182:1255–1256

    Google Scholar 

  9. Christensen BN, Ebner FF (1978) The synaptic architecture of neurones in opossum sensory-motor cortex. J Neurocytol 7:39–60

    Google Scholar 

  10. Cullheim S, Kellerth J-O (1976) Combined light and electron microscopic tracing of neurones, including axons and synaptic terminals, after intracellular injection of horseradish peroxidase. Neuroscience Letters 2:307–313

    Google Scholar 

  11. Cullheim S, Kellerth J-O, Conradi S (1977) Evidence for direct synaptic interconnections between cat spinal α-motoneurones via the recurrent axon collaterals: a morphological study using intracellular horseradish peroxidase. Brain Res 132:1–10

    Google Scholar 

  12. Gillette R, Pomeranz B (1973) Neurone geometry and circuitry via the electron microscope; intracellular staining with osmiophilic polymer. Science 182:1256–1258

    Google Scholar 

  13. Gillette R, Pomeranz B (1975) Ultrastructural correlates of interneuronal function in the abdominal ganglion of Aplysia californica. J Neurobiol 3:463–474

    Google Scholar 

  14. Graham RC, Karnovsky MJ (1966) The early stages of absorption of horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291–302

    Google Scholar 

  15. Hausen K, Wolburg-Buchholz K (1980) An improved cobalt sulphide-silver intensification method for electron microscopy. Brain Res 187:462–466

    Google Scholar 

  16. Hoyle G, Burrows M (1973) Neural mechanisms underlying behaviour in the locust, Schistocerca gregaria. I. Physiology of identified motoneurons in the metathoracic ganglion. J Neurobiol 4:3–41

    Google Scholar 

  17. Itoh K, Konishi A, Nomura S, Mizuno N, Nakamura Y, Sugimoto T (1979) Application of a coupled oxidation reaction to the electron microscopic demonstration of horseradish peroxidase: glucose oxidase method. Brain Res 175:341–346

    Google Scholar 

  18. Jankowska E, Rastad J, Westman J (1976) Intracellular application of horseradish peroxidase and its light and electron microscopical appearance. Brain Res 105:557–562

    Google Scholar 

  19. King DG (1976) Organisation of the crustacean neuropil. II. Distribution of synaptic contacts on identified motor neurones in lobster stomatogastric ganglion. J Neurocytol 5:239–266

    Google Scholar 

  20. King DG (1977) An interneurone in Drosophila synapses within a peripheral nerve onto the dorsal longitudinal muscle motor neurones. Neurose Abs 3:380

    Google Scholar 

  21. Lundquist I, Josefsson J-O (1971) Sensitive method for determination of peroxidase activity in tissue by means of a coupled oxidation reaction. Analyt Biochem 41:567–577

    Google Scholar 

  22. Muller KJ (1979) Synapses between neurones in the central nervous system of the leech. Biol Rev 54:99–134

    Google Scholar 

  23. Muller KJ, McMahan UJ (1976) The shapes of sensory and motor neurones and the distribution of synapses in ganglia of the leech: a study using intracellular injection of horseradish peroxidase. Proc Roy Soc 194:481–499

    Google Scholar 

  24. Pearson KG (1979) Local neurones and local interactions in the nervous systems of invertebrates. In: Schmitt FO, Worden EG (eds) “The Neurosciences, fourth study program”. MIT Press Cambridge Mass and London, pp 145–157

    Google Scholar 

  25. Phillips CE (1980) Intracellularly injected cobaltous ions accumulate at synaptic densities. Science 207:1477–1479

    Google Scholar 

  26. Pitman RM, Tweedle CD, Cohen MJ (1972) Branching of central neurones: intracellular cobalt injection for light and electron microscopy. Science 176:412–414

    Google Scholar 

  27. Pitman RM, Tweedle CD, Cohen MJ (1973) The form of nerve cells: determination by cobalt impregnation. In: Kater SB, Nicholson C (eds) “Intracellular Staining in Neurobiology”. Springer-Verlag, Heidelberg Berlin New York, 83–98

    Google Scholar 

  28. Purves D, McMahan UJ (1972) The distribution of synapses on a physiologically identified motor neurone in the central nervous system of the leech. J Cell Biol 55:205–220

    Google Scholar 

  29. Schrøder HD (1979) Sulfide silver stainability of a type of bouton in spinal cord motoneuron neuropil: an electron microscopic study with Timm's method for the demonstration of heavy metals. J Comp Neurol 186:439–450

    Google Scholar 

  30. Shepherd GM (1979) The synaptic organisation of the brain. Oxford University Press, Oxford and New York

    Google Scholar 

  31. Siegler MVS, Burrows M (1979) The morphology of local non-spiking interneurones in the metathoracic ganglion of the locust. J Comp Neurol 183:121–148

    Google Scholar 

  32. Snow PJ, Ross PK, Brown AG (1976) Tracing axons and axon collaterals of spinal neurones using intracellular injection of horseradish peroxidase. Science 191:312–313

    Google Scholar 

  33. Stewart WW (1978) Functional connections between cells revealed by dye-coupling with highly fluorescent naphthalamide tracer. Cell 14:741–759

    Google Scholar 

  34. Stretton AOW, Kravitz EA (1968) Neuronal geometry: determination with a technique of intracellular dye injection. Science 162:132–134

    Google Scholar 

  35. Tyrer NM, Bell EM (1974) The intensification of cobalt-filled neurone profiles using a modification of Timm's sulphide silver method. Brain Res 73:151–155

    Google Scholar 

  36. Ude J, Agricola H (1978) Synaptic connections of the nervus connectivus in the frontal ganglion of Periplaneta americana. Cell Tiss Res 204:155–159

    Google Scholar 

  37. Usherwood PNR, Grundfest H (1965) Peripheral inhibition in skeletal muscle of insects. J Neurophysiol 28:497–518

    Google Scholar 

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Correspondence to A. H. D. Watson.

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Watson, A.H.D., Burrows, M. Input and output synapses on identified motor neurones of a locust revealed by the intracellular injection of horseradish peroxidase. Cell Tissue Res. 215, 325–332 (1981). https://doi.org/10.1007/BF00239118

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Key words

  • Horseradish peroxidase
  • Locust
  • Motor neurone
  • Synapse
  • Ultrastructure