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Neurofilament protein triplet immunoreactivity in the dorsal root ganglia of the guinea-pig

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Summary

Immunoreactivity for the neurofilament protein triplet was investigated in neurons of the dorsal root ganglia of the guinea-pig by using a battery of antibodies. In unfixed tissue, nearly all neurons in these ganglia demonstrated some degree of neurofilament protein triplet immunoreactivity. Large neurons generally displayed intense immunoreactivity, whereas most small to medium-sized neurons showed faint to moderate immunoreactivity. Double-labelling immunofluorescence demonstrated that most antibodies to the individual subunits of the neurofilament protein triplet had the same distribution and intensity of labelling in sensory neurons. Increasing durations of tissue fixation in aldehyde solutions selectively diminished neurofilament protein triplet immunoreactivity in small to medium-sized neurons. Double-labelling with neurofilament protein triplet antibodies in combination with antibodies to other neuronal markers, such as neuron-specific enolase, substance P and tyrosine hydroxylase, showed that tissue processing conditions affect the degree of co-localization of immunoreactivity to the neurofilament protein triplet and to these other neuronal markers. These results indicate that, with a judicious manipulation of the duration of tissue fixation, neurofilament protein triplet immunoreactivity can be used in combination with other neuronal markers to distinguish groups of neurons according to their size and chemical coding.

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References

  • Anderton B, Coakham HB, Garson JA, Harper EI, Lawson SN (1982) A monoclonal antibody against neurofilament protein specifically labels the large light cell population in rat dorsal root ganglia. J Physiol (Lond) 334:97–98P

    Google Scholar 

  • Autilio-Gambetti L, Crane R, Gambetti P (1986) Binding of Bodian's silver and monoclonal antibodies to defined regions of human neurofilament subunits: Bodian's silver reacts with a highly charged unique domain of neurofilaments. J Neurochem 46:366–370

    Google Scholar 

  • Berglund AM, Ryugo DK (1986) A monoclonal antibody labels type II neurons in the spiral ganglion. Brain Res 383:327–332

    Google Scholar 

  • Bishop AE, Carlei F, Lee V, Trojanowski J, Marangos PJ, Dahl D, Polak JM (1985) Combined immunostaining of neurofilaments, neuron specific enolase, GFAP, and S-100. Histochemistry 82:93–97

    Google Scholar 

  • Björklund H, Dahl D, Seiger Å (1984) Neurofilament and glial fibrillary acid protein-related immunoreactivity in the rodent enteric nervous system. Neuroscience 12:277–287

    Google Scholar 

  • Campbell MJ, Morrison JH (1989) Monoclonal antibody to neurofilament protein (SMI-32) labels a subpopulation of pyramidal neurons in the human and monkey neocortex. J Comp Neurol 282:191–205

    Google Scholar 

  • Costa M, Furness JB, Gibbins IL (1986) Chemical coding of enteric neurons. Prog Brain Res 68:217–239

    Google Scholar 

  • Dräger UC, Hofbauer A (1984) Antibodies to heavy neurofilament subunit detect a subpopulation of damaged ganglion cells in retina. Nature 309:624–626

    Google Scholar 

  • Duce IR, Keen P (1977) An ultrastructural classification of the neuronal cell bodies of the rat dorsal root ganglion using zinc iodide-osmium impregnation. Cell Tissue Res 185:263–277

    Google Scholar 

  • Gambetti P, Autilio-Gambetti L, Papasozomenos SC (1981) Bodian's silver method stains neurofilament polypeptides. Science 213:1521–1522

    Google Scholar 

  • Gibbins IL, Furness JB, Costa M (1987) Pathway-specific patterns of co-existence of substance P, calcitonin gene-related peptide, cholecystokinin and dynorphin in neurons of the dorsal root ganglia of the guinea-pig. Cell Tissue Res 248:417–437

    Google Scholar 

  • Gibson SJ, Polak JM, Bloom SR, Sabate IM, Muldberry PM, Ghatei MA, McGregor CP, Morrison JFB, Kelly JS, Evans RM, Rosenfeld MG (1984) Calcitonin gene-related peptide immunoreactivity in the spinal cord of man and of eight other species. J Neurosci 4:3101–3111

    Google Scholar 

  • Goldstein ME, Cooper HS, Bruce J, Carden MJ, Lee VM-Y, Schlaepfer WW (1987) Phosphorylation of neurofilament proteins and chromatolysis following transection of rat sciatic nerve. J Neurosci 7:1586–1594

    Google Scholar 

  • Gray EG, Guillery RW (1961) The basis of silver staining of synapses of mammalian spinal cord: a light and electron microscope study. J Physiol (Lond) 157:581–588

    Google Scholar 

  • Hickey WF, Lee V, Trojanowski JQ, McMillan LJ, McKearn TJ, Gonatas J, Gonatas NK (1983) Immunohistochemical application of monoclonal antibodies against myelin basic protein and neurofilament triplet protein subunits. J Histochem Cytochem 31:1126–1135

    Google Scholar 

  • Hökfelt T, Millhorn D, Seroogy K, Tsuro Y, Ceccatelli S, Lindh B, Meister B, Melander T, Schalling M, Bartfai T, Terenius L (1987) Coexistence of peptides with classical neurotransmiters. Experientia 43:768–780

    Google Scholar 

  • Hua X-Y, Theodorsson-Norheim E, Brodin E, Lundberg JM, Hökfelt T (1985) Multiple tachykinins (neurokinin A, neuropeptide K and substance P) in capsaicin-sensitive sensory neurons in the guinea-pig. Regul Pept 13:1–19

    Google Scholar 

  • Kai-Kai MA, Anderton BH, Keen P (1986) A quantitative analysis of the interrelationships between subpopulations of rat sensory neurones containing arginine vasopressin or oxytocin and those containing substance P, fluoride-resistant acid phosphatase of neurofilament protein. Neuroscience 18:465–486

    Google Scholar 

  • Kummer W, Gibbins IL, Steffan P, Kapoor V (1990) Catecholamines and catecholamine synthesizing enzymes in guinea-pig sensory ganglia. Cell Tissue Res 261:595–606

    Google Scholar 

  • Lawson SN, Waddel PJ (1985) The antibody RT97 distinguishes between sensory cell bodies with myelinated and unmyelinated peripheral processes in the rat. J Physiol (Lond) 371: 59P

  • Lawson SN, Harper EI, Harper AA, Garson JA, Anderton BH (1984) A monoclonal antibody against neurofilament protein specifically labels a subpopulation of rat sensory neurones. J Comp Neurol 228:263–272

    Google Scholar 

  • Lindh B, Dalsgaard C-J, Elfvin L-G, Hökfelt T, Cuello AC (1983) Evidence of substance P immunoreactive neurons in the dorsal root ganglia and vagal ganglia projecting to the guinea pig pylorus. Brain Res 269:365–369

    Google Scholar 

  • Lundberg JM, Hökfelt T, Nilsson G, Terenius L, Rehfeld J, Elde R, Said S (1978) Peptide neurons in the vagus, splachnic and sciatic nerves. Acta Physiol Scand 104:499–501

    Google Scholar 

  • McCarthy PW, Lawson SN (1989) Cell type and conduction velocity of rat primary sensory neurons with substance P-like immunorcactivity. Neuroscience 28:745–753

    Google Scholar 

  • Millard CL, Woolf CJ (1988) Sensory innervation of the hairs of the rat hindlimb: a light microscope analysis. J Comp Neurol 277:183–194

    Google Scholar 

  • Morrison JH, Lewis DA, Campbell MJ, Huntley GW, Benson DL, Bouras C (1987) A monoclonal antibody to non-phosphorylated neurofilament protein marks the vulnerable cortical neurons in Alzheimer's disease. Brain Res 416:331–336

    Google Scholar 

  • Nohr D, Weihe E, Zentel HJ, Arendt RM (1989) Atrial natriuretic factor-like immunoreactivity in spinal cord and in primary sensory neurons of spinal and trigeminal ganglia of guinea-pig: correlation with tachykinin immunoreactivity. Cell Tissue Res 258:387–392

    Google Scholar 

  • Poltorak M, Freed WJ (1989) Immunoreactive phosphorylated epitopes on neurofilaments in neuronal perikarya may be obscured by tissue preprocessing. Brain Res 480:349–354

    Google Scholar 

  • Price J (1985) An immunohistochemical and quantitative examination of dorsal root ganglion neuronal subpopulations. J Neurosci 5:555–563

    Google Scholar 

  • Romand R, Hafidi A, Despres G (1988) Immunocytochemical localization of neurofilament subunits in the spiral ganglion of the adult rat. Brain Res 462:167–173

    Google Scholar 

  • Seiger Å, Dahl D, Ayer-LeLievre C, Björklund H (1984) Appearance and distribution of neurofilament immunoreactivity in iris nerves. J Comp Neurol 223:457–470

    Google Scholar 

  • Sharp GA, Shaw G, Weber K (1982) Immunoelectronmicroscopical localization of the three neurofilament triplet proteins along neurofilaments of cultured dorsal root ganglion neurones. Exp Cell Res 137:403–413

    Google Scholar 

  • Shaw G, Weber K (1983) The structure and development of the rat retina: an immunofluorescence microscopical study using antibodies specific for intermediate filament proteins. Eur J Cell Biol 30:219–232

    Google Scholar 

  • Shaw G, Osborn M, Weber K (1981) Arrangement of neurofilaments, microtubules and microfilament-associated proteins in cultured dorsal root ganglia cells. Eur J Cell Biol 24:20–27

    Google Scholar 

  • Steinert PM, Roop DR (1988) Molecular and cellular biology of intermediate filaments. Ann Rev Biochem 57:593–625

    Google Scholar 

  • Trojanowski JQ, Walkenstein N, Lee VM-Y (1986) Expression of neurofilament subunits in neurons of the central and peripheral nervous system: an immunohistochemical study with monoclonal antibodies. J Neurosci 6:650–660

    Google Scholar 

  • Vickers JC, Costa M, Vitadello M, Dahl D, Marotta CA (1990) Neurofilament protein triplet immunoreactivity in distinct subpopulations of peptide-containing neurons in the guinea-pig coeliac ganglion. Neuroscience 39:743–759

    Google Scholar 

  • Vitadello M, Triban C, Fabris M, Donà M, Gorio A, Schiaffino S (1987) A developmentally regulated isoform of 150 000 molecular weight neurofilament protein specifically expressed in autonomic and small sensory neurons. Neuroscience 23:931–941

    Google Scholar 

  • Weihe E, Hartschuh W, Weber E (1985) Prodynorphin opioid peptides in small somatosensory primary afferents of guinea pig. Neurosci Lett 58:347–352

    Google Scholar 

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Authors and Affiliations

  1. Centre for Neuroscience, Department of Physiology, Flinders University of South Australia, GOP Box 2100, 5001, Adelaide, Australia

    James C. Vickers & Marcello Costa

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  1. James C. Vickers
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  2. Marcello Costa
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Vickers, J.C., Costa, M. Neurofilament protein triplet immunoreactivity in the dorsal root ganglia of the guinea-pig. Cell Tissue Res 265, 159–167 (1991). https://doi.org/10.1007/BF00318150

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  • DOI: https://doi.org/10.1007/BF00318150

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