Skip to main content
Log in

Ultrastructural studies of the myenteric plexus and smooth muscle in organotypic cultures of the guinea-pig small intestine

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

External muscle and myenteric plexus from the small intestine of adult guinea-pigs were maintained in vitro for 3 or 6 days. Myenteric neurons and smooth muscle cells from such organotypic cultures were examined at the electron-microscopic level. An intact basal lamina was found around the myenteric ganglia and internodal strands. Neuronal membranes, nuclei and subcellular organelles appeared to be well preserved in cultured tissues and ribosomes were abundant. Dogiel type-II neurons were distinguishable by their elongated electron-dense mitochondria, numerous lysosomes and high densities of ribosomes. Vesiculated nerve profiles contained combinations of differently shaped vesicles. Synaptic membrane specializations were found between vesiculated nerve profiles and nerve processes and cell bodies. The majority of nerve fibres were well preserved in the myenteric ganglia, in internodal strands and in bundles running between circular muscle cells. No detectable changes were found in the ultrastructure of the somata and processes of glial cells. Longitudinal and circular muscle cells from cultured tissue had clearly defined membranes with some close associations with neighbouring muscle cells. Caveolae occurred in rows that ran parallel to the long axis of the muscle cells. These results indicate that the ultrastructural features of enteric neurons and smooth muscle of the guinea-pig small intestine are well preserved in organotypic culture.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Baluk P, Jessen KR, Saffrey MJ, Burnstock G (1983) The enteric nervous system in tissue culture. II. Ultrastructural studies of cell types and their relationships. Brain Res 262:37–47

    Google Scholar 

  • Berthoud H-R, Powley TL (1992) Vagal afferent innervation of the rat fundic stomach: morphological characterization of the gastric tension receptor. J Comp Neurol 319:261–276

    Google Scholar 

  • Bornstein JC, North RA, Costa M, Furness JB (1984) Excitatory synaptic potentials due to activation of neurons with short projections in the myenteric plexus. Neuroscience 11:723–731

    Google Scholar 

  • Brookes SJH, Costa M (1990) Identification of enteric motor neurons which innervate the circular muscle of the guinea pig small intestine. Neurosci Lett 118:227–230

    Google Scholar 

  • Brookes SJH, Steele PA, Costa M (1991) Identification and immunohistochemistry of cholinergic and non-cholinergic circular muscle motor neurones in the guinea-pig small intestine. Neuroscience 42:863–878

    Google Scholar 

  • Brookes SJH, Song Z-M, Steele PA, Costa M (1992) Identification of motor neurons to the longitudinal muscle of the guinea pig ileum. Gastroenterology 103:961–973

    Google Scholar 

  • Brookes SJH, Ramsay GA, Song Z-M, Costa M (1993) Aboral projections of putative sensory neurones in the guinea-pig small intestine. J Gastroenterol Hepathol 8:A29

    Google Scholar 

  • Cook R, Burnstock G (1976a) The ultrastructure of Auerbach's plexus in the guinea-pig. I. Neuronal elements. J Neurocytol 5:174–194

    Google Scholar 

  • Cook R, Burnstock G (1976b) The ultrastructure of Anerbach's plexus in the guinea-pig. II. Non-neuronal elements. J Neurocytol 5:195–206

    Google Scholar 

  • Dogiel AS (1899) Ueber den Bau der Ganglien in den Geflechten des Darmes und der Gallenblase des Menschen und der Säugethiere. Arch Anat Physiol Leipzig, Anat Abt:130–158

  • Dreyfus CF, Bornstein MB (1977) Synthesis of serotonin by neurons of the myenteric plexus in situ and in organotypic tissue culture. Brain Res 128:125–139

    Google Scholar 

  • Dreyfus CF, Sherman DL, Gershon MD (1977) Uptake of serotonin by intrinsic neurons of the myenteric plexus grown in organotypic tissue culture. Brain Res 128:109–123

    Google Scholar 

  • Furness JB, Costa M (1987) The enteric nervous system. Churchill Livingstone, Edinburgh

    Google Scholar 

  • Furness JB, Llewellyn-Smith IJ, Bornstein JC, Costa M (1988) Chemical neuroanatomy and the analysis of neuronal circuitry in the enteric nervous system. In: Björklund A, Hökfelt T, Owman C (eds) The peripheral nervous system. Handbook of chemical neuroanatomy, vol 6. Elsevier, Amsterdam, pp 161–218

    Google Scholar 

  • Gabella G (1971) Caveolae intracellulares and sarcoplasmic reticulum in smooth muscle. J Cell Sci 8:601–609

    Google Scholar 

  • Gabella G (1972) Fine structure of the myenteric plexus in the guinea-pig ileum. J Anat 111:69–97

    Google Scholar 

  • Gabella G (1987) Structure of muscles and nerves in the gastrointestinal tract. In: Johnson LR (ed) Physiology of the gastrointestinal tract, 2nd edn. Raven Press, New York, pp 335–381

    Google Scholar 

  • Grobstein C (1965) Differentiation: environmental factors, chemical and cellular. In: Willmer EN (ed) Cells and tissues in culture, methods, biology and physiology, vol 1. Academic Press, London New York, pp 463–488

    Google Scholar 

  • Hanani M, Baluk P, Burnstock G (1982) Myenteric neurons express electrophysiological and morphological diversity in tissue culture. J Auton Nerv Syst 5:155–164

    Google Scholar 

  • Jessen KR, Burnstock G (1982) The enteric nervous system in tissue culture: a new mammalian model for the study of complex nervous networks. In: Kalsner S (ed) Trends in autonomic pharmacology, vol 2. Urban and Schwarzenberg, Baltimore, pp 95–115

    Google Scholar 

  • Jessen KR, McConnell D, Purves RD, Burnstock G, Chamley-Campbell J (1978) Tissue culture of mammalian enteric neurons. Brain Res 152:573–579

    Google Scholar 

  • Jessen KR, Saffrey MJ, Van Noorden S, Bloom SR, Polak JM, Burnstock G (1980) Immunohistochemical studies of the enteric nervous system in tissue culture and in situ: localization of vasoactive intestinal polypeptide (VIP), substance-P and enkephalin immunoreactive nerves in the guinea-pig gut. Neuroscience 5:1717–1735

    Google Scholar 

  • Jessen KR, Saffrey MJ, Burnstock G (1983a) The enteric nervous system in tissue culture. I. Cell types and their interactions in explants of the myenteric and submucous plexuses from guinea pig, rabbit and rat. Brain Res 262:17–35

    Google Scholar 

  • Jessen KR, Saffrey MJ, Baluk P, Hanani M, Burnstock G (1983b) The enteric nervous system in tissue culture. III. Studies on neuronal survival and retention of biochemical and morphological differentiation. Brain Res 262:49–62

    Google Scholar 

  • Kow L, Brookes SJH, Costa M (1993) Enteric reflexes in organotypic culture of guinea-pig small intestine. Proc Aust Neurosci Soc 4:79

    Google Scholar 

  • Lewis WH, Lewis MR (1912) The cultivation of the sympathetic nerves from the intestine of chick-embryos in saline solutions. Anat Rec 6:7–30

    Google Scholar 

  • Llewellyn-Smith IJ, Costa M, Furness JB (1985) Light and electron microscopic immunocytochemistry of the same nerves from whole-mount preparations. J Histochem Cytochem 33:857–866

    Google Scholar 

  • Nishi R, Willard AL (1985) Neurons dissociated from rat myenteric plexus retain differentiated properties when grown in cell culture. I. Morphological properties and immunocytochemical localization of transmitter candidates. Neuroscience 16:187–199

    Google Scholar 

  • O'Neal JT, Westrum LE (1973) The fine structural synaptic organization of the cat lateral cuneate nucleus. A study of sequential alterations in degeneration. Brain Res 51:97–124

    Google Scholar 

  • Pompolo S, Furness JB (1988) Ultrastructure and synaptic relationships of calbindin-reactive, Dogiel type-II neurons, in myenteric ganglia of guinea-pig small intestine. J Neurocytol 17:771–782

    Google Scholar 

  • Raisman G (1973) Electron microscopic studies of the development of new neurohaemal contacts in the median eminence of the rat after hypophysectomy. Brain Res 55:245–261

    Google Scholar 

  • Rikimaru A (1971) Contractile properties of organ-cultured intestinal smooth muscle. Tohoku J Exp Med 103:317–329

    Google Scholar 

  • Rush RA, Saltis J, Smet PJ, Williams R (1992) Survival factors for autonomic neurons. In: Hendry IA, Hill CE (eds) Development, regeneration and plasticity of the autonomic nervous system. Harwood Academic, Reading, UK, pp 305–362

    Google Scholar 

  • Saffrey MJ, Burnstock G (1988) Peptide-containing neurons in explant cultures of guinea-pig myenteric plexus during development in vitro: gross morphology and growth patterns. Cell Tissue Res 254:167–176

    Google Scholar 

  • Saffrey MJ, Bailey DJ, Burnstock G (1991) Growth of enteric neurones from isolated myenteric ganglia in dissociated cell culture. Cell Tissue Res 265:527–534

    Google Scholar 

  • Saffrey MJ, Hassall CJ, Hoyle CH, Belai A, Moss J, Schmidt HH, Forstermann U, Murad F, Burnstock G (1992) Colocalization of nitric oxide synthase and NADPH-diaphorase in cultured myenteric neurones. Neuroreport 3:333–336

    Google Scholar 

  • Song Z-M, Brookes SJH, Costa M (1991) Identification of myenteric neurons which project to the mucosa of the guinea-pig small intestine. Neurosci Lett 129:294–298

    Google Scholar 

  • Song Z-M, Brookes SJH, Steele PA, Costa M (1992) Projections and pathways of submucous neurons to the mucosa of the guinea-pig small intestine. Cell Tissue Res 269:87–98

    Google Scholar 

  • Song Z-M, Brookes SJH, Costa M (1993) Identification of a subpopulation of anally-directed calbindin-immunoreactive neurons in the myenteric plexus of the guinea-pig small intestine. Proc Aust Neurosci Soc 4:76

    Google Scholar 

  • Stach W (1989) A revised morphological classification of neurons in the enteric nervous system. In: Singer MV, Goebell H (eds) Nerves and the gastrointestinal tract. Kluwer Academic, Lancaster, pp 29–45

    Google Scholar 

  • Willard AL (1990) Substance P mediates synaptic transmission between rat myenteric neurones in cell culture. J Physiol (Lond) 426:453–471

    Google Scholar 

  • Willard AL, Nishi R (1985a) Neurons dissociated from rat myenteric plexus retain differentiated properties when grown in cell culture. II. Electrophysiological properties and responses to neurotransmitter candidates. Neuroscience 16:201–211

    Google Scholar 

  • Willard AL, Nishi R (1985b) Neurons dissociated from rat myenteric plexus retain differentiated properties when grown in cell culture. III. Synaptic interactions and modulatory effects of neurotransmitter candidates. Neuroscience 16:213–221

    Google Scholar 

  • Yanase J (1907) Beiträge zur Physiologie der peristaltischen Bewegungen des embryonalen Darmes. Pflügers Arch 117:345

    Google Scholar 

  • Zhou X-F, Rush RA (1993) Localization of neurotrophin-3-like immunoreactivity in peripheral tissues of the rat. Brain Res 621:189–199

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Song, ZM., Brookes, S.J.H., Llewellyn-Smith, I.J. et al. Ultrastructural studies of the myenteric plexus and smooth muscle in organotypic cultures of the guinea-pig small intestine. Cell Tissue Res 280, 627–637 (1995). https://doi.org/10.1007/BF00318365

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00318365

Key words

Navigation