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The survival of neonatal rat retinal ganglion cells in vitro is enhanced in the presence of appropriate parts of the brain

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Summary

The enzyme horseradish peroxidase (HRP) was injected into the visual centres of the brains of neonatal rats. Following dissociation of retinae into tissue culture, the ganglion cells could be identified by appropriate histochemical staining for HRP reaction product. Cultures were prepared of dissociated retinae from rats aged 2–6 days postnatal. After 3 h the cultures were fixed, and HRP-labelled cells visualized and counted. Estimates were made of the number of ganglion cells per retina at each age. Results indicated a loss of ganglion cells during the first few postnatal days. This loss paralleled that observed in vivo. It was further found the retinal ganglion cells died rapidly in vitro when cultured in a minimal medium. Only 50% of ganglion cells originally plated remained viable after 24 h. However, the survival rate could be increased to 100% by coculturing the cells with diencephalon and mesencephalon; these contain the retinorecipient nuclei. Coculturing with cerebellum did not result in such an enhanced survival rate. Ganglion cells could be maintained over longer periods of time by reinoculating the cultures with additional tissue containing diencephalon and mesencephalon. These results support the hypothesis that developing neurons require trophic factors from their target tissues in order to survive.

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References

  • Abols IA, Basbaum AI (1979) The posterior pretectal nucleus: Evidence for a direct projection to the inferior olive of the cat. Neurosci Lett 13: 111–116

    Google Scholar 

  • Azzena GB, Mameli O, Tolu E (1980) Distribution of visual input to the vestibular nuclei. Arch Ital Biol 118: 196–204

    Google Scholar 

  • Bennett MR (1981) Development of the neuromuscular synapse. Invited lecture. Proc Aust Physiol Pharmacol Soc 12: 41–58

    Google Scholar 

  • Bennett MR, White W (1979) The survival and development of cholinergic neurons in potassium-enriched media. Brain Res 173: 549–553

    Google Scholar 

  • Benowitz LI, Shashoua VE (1979) Immunoreactive sites for nerve growth factor (NGF) in the goldfish brain. Brain Res 172: 561–565

    Google Scholar 

  • Bonhoeffer F, Huf J (1980) Recognition of cell types by axonal growth cones in vitro. Nature 288: 162–164

    Google Scholar 

  • Brown JT, Chan Palay J, Palay SL (1977) A study of afferent input to the inferior olivary complex in rat by retrograde axonal transport of horseradish peroxidase. J Comp Neurol 197: 1–22

    Google Scholar 

  • Bunt AH, Lund RD, Lund JS (1974) Retrograde axonal transport of horseradish peroxidase by ganglion cells of the albino rat. Brain Res 73: 215–228

    Google Scholar 

  • Combes PC, Privat A, Pessac B, Calothy G (1977) Differentiation of chick embryo neuroretina cells in monolayer cultures. An ultrastructural study. Cell Tissue Res 185: 159–173

    Google Scholar 

  • Cowan WM (1973) Neuronal death as a regulative mechanism in the control of cell number in the nervous system. In: Rockstein M (ed) Development and aging in the nervous system. Academic Press, New York, pp 19–41

    Google Scholar 

  • Cowan WM (1979) Selection and control in neurogenesis. In: Schmitt FO, Worden FG (eds) The neurosciences fourth study program. MIT Press, Cambridge, MA, 59–79

    Google Scholar 

  • Cunningham TJ, Mohler IM, Giordano DL (1981) Naturally occurring neuron death in the ganglion cell layer of the neonatal rat: Morphology and evidence for regional correspondence with death in superior colliculus. Dev Brain Res 2: 203–215

    Google Scholar 

  • Green LA (1974) A dissociated cell culture bioassay for nerve growth factor. Neurobiol 4: 286–292

    Google Scholar 

  • Hanker JS, Yates PE, Metz CB, Rustioni A (1977) A new specific sensitive and non-carcinogenic reagent for the demonstration of horseradish peroxidase. Histochem J 9: 789–792

    Google Scholar 

  • Hughes WF, LaVelle A (1975) The effects of early tectal lesions on development in the retinal ganglion cell layer of chick embryos. J Comp Neurol 163: 265–284

    Google Scholar 

  • Hughes WF, McLoon SC (1979) Ganglion cell death during normal retinal development in the chick: Comparisons with cell death induced by early target field destruction. Exp Neurol 66: 587–601

    Google Scholar 

  • Jeffrey G, Perry VH (1981) Evidence for ganglion cell death during development of the ipsilateral retinal projection in the rat. Dev Brain Res 2: 176–180

    Google Scholar 

  • Karlsson U (1967) Three-dimensional studies of neurons in the lateral geniculate nucleus of the rat. III. Specialized neuronal contacts in the neuropil. J Ultrastruct Res 17: 137–157

    Google Scholar 

  • Keefer DA (1978) Horseradish peroxidase as a retrogradely transported detailed dendritic marker. Brain Res 140: 15–32

    Google Scholar 

  • Lam K, Sefton AJ, Bennett MR (1982) Loss of axons from the optic nerve of the rat during early postnatal development. Dev Brain Res 3: 487–491

    Google Scholar 

  • Landa KB, Adler R, Manthorpe M, Varon S (1980) Cholinergic neuronotrophic factors. III. Developmental increase of trophic activity for chick embryo ciliary ganglion neurons in their intraocular target tissues. Dev Biol 74: 401–408

    Google Scholar 

  • LaVail JH, Cowan WM (1971) The development of the chick optic tectum. I. Normal morphology and cytoarchitectonic development. Brain Res 28: 391–419

    Google Scholar 

  • Lund RD, Bunt AH (1976) Prenatal development of central optic pathways in albino rats. J Comp Neurol 165: 247–264

    Google Scholar 

  • Lund RD, Lund RS (1972) Development of synaptic patterns in the superior colliculus of the rat. Brain Res 42: 1–20

    Google Scholar 

  • McLoon SC, Lund RD (1980a) Specific projections of retina transplanted to rat brain. Exp Brain Res 40: 273–282

    Google Scholar 

  • McLoon SC, Lund RD (1980b) Identification of cells in retinal transplants which project to host visual centres: A horseradish peroxidase study in rats. Brain Res 197: 491–495

    Google Scholar 

  • Mesulam MM (1978) Tetramethylbenzidine for horseradish peroxidase neurohistochemistry. Non-carcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents. J Histochem Cytochem 26: 106–117

    CAS  PubMed  Google Scholar 

  • Michaelson IC (1954) Retinal circulation in man and animals. Thomas, Illinois, pp 64–73

    Google Scholar 

  • Nurcombe V, Bennett MR (1981) Embryonic chick retinal ganglion cells identified in vitro. Their survival is dependent on a factor from the optic tectum. Exp Brain Res 44: 249–258

    Google Scholar 

  • Perry VH, Cowey A (1979) The effects of unilateral cortical and tectal lesions on retinal ganglion cells in rats. Exp Brain Res 35: 85–95

    Google Scholar 

  • Potts RA, Dreher B, Bennett MR (1982) The loss of ganglion cells in the developing retina of the rat. Dev Brain Res 3: 481–486

    Google Scholar 

  • Rager G (1978) Systems-matching by degeneration. II. Interpretation of the generation and degeneration of retinal ganglion cells in the chicken by a mathematical model. Exp Brain Res 33: 79–90

    Google Scholar 

  • Rager G, Rager U (1978) Systems-matching by degeneration. I. A quantitative electron microscopic study of the generation and degeneration of retinal ganglion cells in the chicken. Exp Brain Res 33: 65–78

    Google Scholar 

  • Rubin NA, Tarsio JF, Borthwick AC, Gregerson DS, Reid TW (1981) Identification and characterization of a growth factor secreted by an established cell line of human retinoblastoma maintained in serum-free medium. Vision Res 21: 105–112

    Google Scholar 

  • Schwartz M, Agranoff BW (1981) Outgrowth and maintenance of neurites from cultured goldfish retinal ganglion cells. Brain Res 206: 331–343

    Google Scholar 

  • Sengelaub DR, Finlay BL (1981) Early removal of one eye reduces normally occurring cell death in the remaining eye. Science 213: 573–574

    Google Scholar 

  • Smalheiser NR, Crain SM, Bornstein MB (1981a) Development of ganglion cells and their axons in organized cultures of foetal mouse expiants. Brain Res 204: 159–178

    Google Scholar 

  • Smalheiser NR, Peterson ER, Crain SM (1981b) Neurites from mouse retina and dorsal root ganglion expiants show specific behaviour within co-cultured tectum or spinal cord. Brain Res 208: 499–505

    Google Scholar 

  • Stach RW, Stach BM, West NR (1979) Nerve fibre outgrowth from dorsal root ganglia: Ion dependency of nerve growth factor action. J Neurochem 33: 845–855

    Google Scholar 

  • Turner JE, Delaney RK, Johnson JE (1981) Retinal ganglion cell response to axotomy and nerve growth factor antiserum treatment in the regenerating visual system of the goldfish: An in vivo and in vitro analysis. Brain Res 204: 283–294

    Google Scholar 

  • Varon S, Bunge RP (1978) Trophic mechanisms in the peripheral nervous system. Ann Rev Neurosci 1: 327–361

    Google Scholar 

  • Winfield JA, Hendrickson A, Kimm J (1978) Anatomical evidence that the medial terminal nucleus of the accessory optic tract in mammals provides a visual mossy fibre input to the flocculus. Brain Res 151: 175–182

    Google Scholar 

  • Winick M, Greenberg RE (1965) Appearance and distribution of a nerve-growth-promoting protein during development. Pediatrics 35: 221–228

    Google Scholar 

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Authors and Affiliations

  1. Neurobiology Research Centre, and the Dept. of Anatomy, University of Sydney, 2001, Sydney, Australia

    C. A. McCaffery, M. R. Bennett DSc & B. Dreher

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  1. C. A. McCaffery
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  2. M. R. Bennett DSc
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  3. B. Dreher
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Supported by the O.P.S.M. Research Foundation, the A.R.G.C. and the N.H. & M.R.C.

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McCaffery, C.A., Bennett, M.R. & Dreher, B. The survival of neonatal rat retinal ganglion cells in vitro is enhanced in the presence of appropriate parts of the brain. Exp Brain Res 48, 377–386 (1982). https://doi.org/10.1007/BF00238614

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  • DOI: https://doi.org/10.1007/BF00238614

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