Summary
The distribution of phosphorylated and nonphosphorylated neurofilament epitopes was determined immunocytochemically in adjacent 2 μm-thick sections of sciatic nerve, ventral root and spinal cord. Staining was scored as either intense, moderate or absent and the proportion of labeled axons was calculated for each category. Nearly all sciatic nerve and ventral root axons were immunoreactive with both antibodies against phosphorylated and non-phosphorylated neurofilaments and there were no significant differences in the number of intensely- or moderately-labeled axons. Within the spinal cord however, while the majority of large caliber axons was stained with both antibodies, there was a significant number of small caliber axons which stained only with antibodies against phosphorylated neurofilaments. These results show that phosphorylated and nonphosphorylated neurofilaments are extensively codistributed in CNS and PNS axons, and that in the CNS, staining intensity for non-phosphorylated epitopes is less in the smaller axons.
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References
Bignami A, Chi NH, Dahl D (1986) Neurofilament phosphorylation in peripheral nerve regeneration. Brain Res 375:73–82
Black MM, Lee VM-Y (1988) Phosphorylation of neurofilament proteins in intact neurons: demonstration of phosphorylation in cell bodies and axons. J Neurosci 8:3296–3305
Breen KC, Robinson PA, Wion D, Anderton BH (1988) Partial sequence of the rat heavy neurofilament polypeptide (NF-H): Identification of putative phosphorylation sites. FEBS Lett 214:213–218
Carden MJ, Schlaepfer WW, Lee VM-Y (1985) The structure, biochemical properties and immunogenicity of neurofilament peripheral regions are determined by phosphorylation state. J Biol Chem 260:9805–9817
Dahl D (1983) Immunohistochemical differences between neurofilaments in perikarya, dendrites and axons. Exp Cell Res 149:397–408
Dahl D (1987) Maturation of a large neurofilament protein (NF 150K) in rat postnatal development. J Neurosci Res 17:367–374
Dahl D, Bignami A (1986) Neurofilament phosphorylation in development. A sign of maturation? Exp Cell Res 162:220–230
Dahl D, Labkovsky B, Bignami A (1988) Neurofilament phosphorylation in axons and perikarya: immunofluorescence study of the rat spinal cord and dorsal root ganglia with monoclonal antibodies. J Comp Neurol 271:445–450
Drager UC, Hofbauer A (1984) Antibodies to heavy neurofilament subunit detect a subpopulation of damaged ganglion cells in retina. Nature 309:624–626
Foster GA, Dahl D, Lee VM-Y (1987) Temporal and topographic relationships between the phosphorylated and non-phosphorylated epitopes of the 200 kDa neurofilament proteins during development in vitro. J Neurosci 7:2651–2663
Geisler N, Fischer S, Vandekerckhove J, Vandomme J, Plessmann V, Weber K (1985) Protein-chemical characterization of NF-H, the largest mammalian neurofilament component—Intermediate filament-type sequences followed by a unique carboxy-terminal extension. EMBO J 4:57–63
Glicksman MA, Soppet D, Willard MB (1987) Posttranslational modification of neurofilament polypeptides in rabbit retina. J Neurobiol 18:167–196
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
Hirokawa N (1982) Cross-linker system between neurofilament, microtubules and membrane organelles in frog axons, revealed by the quick-freeze, deep-etching method. J Cell Biol 94:129–142
Hirokawa N, Glicksman MA, Willard M (1984) Organization of mammalian neurofilament polypeptides within the neuronal cytoskeleton. J Cell Biol 98:1523–1536
Hoffman PN, Lasek RJ (1975) The slow component of axonal transport. Identification of major structural polypeptides of the axon and their generality among mammalian neurons. J Cell Biol 66:351–366
Hoffman PN, Thompson GW, Griffin JW, Price DL (1985) Changes in neurofilament transport coincide temporally with alterations in the caliber of axons in regenerating motor fibers. J Cell Biol 101:1332–1340
Julien J-P, Cote F, Beaudet L, Sidky M, Flavell D, Grosveld F, Muchynski W (1988) Sequence and structure of the mouse gene coding for the largest neurofilament subunit. Gene 68:307–314
Lee VM-Y, Carden MJ, Schlaepfer WW, Trojanowski JQ (1987) Monoclonal antibodies distinguish several differentially phosphorylated states of the two largest rat neurofilament subunits (NF-H and NF-M) and demonstrate their existence in the normal nervous system of adult rats. J Neurosci 7:3474–3488
Lees JF, Shneidman PS, Skuntz SF, Carden MJ, Lazzarini RA (1988) The structure and organization of the human heavy neurofilament subunit (NF-H) and the gene encoding it. EMBO J 7:1947–1955
Lewis SE, Nixon RA (1988) Multiple phosphorylated variants of the high molecular mass subunit of neurofilaments in axons of retinal cell neurons: Characterization and evidence for their differential association with stationary and moving neurofilaments. J Cell Biol 107:2689–2701
Matus A (1988) Neurofilament protein phosphorylation—where, when and why. Trends Neurosci 11:291–292
McQuarrie IG (1983) Role of the axonal cytoskeleton in the regenerating nervous system. In: Seil FJ (ed) Nerve, organ and tissue regeneration: Research Perspectives. Academic Press, New York, pp 51–88
Munoz DG, Greene C, Perl DP, Sekoe DJ (1988) Accumulation of phosphorylated neurofilaments in anterior horn motoneurons of amyotrophic lateral sclerosis patients. J Neuropathol Exp Neurol 47:9–18
Oblinger MM (1987) Characterization of posttranslational processing of the mammalian high-molecular weight neurofilament protein in vivo. J Neurosci 7:2510–2521
Rhoades RW, Hsu L, Parfett G (1979) An electron microscopic analysis of the optic nerve in the golden hamster. J Comp Neurol 186:491–504
Rosenfeld J, Dorman ME, Griffin JW, Sternberger LA, Sternberger NH, Price DL (1987) Distribution of neurofilament antigens after axonal injury. J Neuropathol Exp Neurol 46:269–282
Schmidt ML, Carden MJ, Lee VM-Y, Trojanowski JQ (1987) Phosphate dependent and independent neurofilament epitopes in the axonal swellings of patients with motor neuron disease and controls. Lab Invest 56:282–294
Sharp GA, Shaw G, Weber K (1982) Immunoelectron microscopical localization of the three neurofilament triplet proteins along neurofilaments of cultured dorsal root ganglion neurons. Exp Cell Res 137:403–413
Sloan KE, Stevenson JA (1987) Differential distribution of phosphorylated and non-phosphorylated neurofilaments within the retina and optic nerve of hamsters. Brain Res 437:365–368
Sternberger LA, Sternberger NH (1983) Monoclonal antibodies distinguish phosphorylated and nonphosphorylated forms of neurofilaments in situ. Proc Natl Acad Sci USA 80:6126–6130
Sternberger LA, Hardy PH, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase anti-horseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333
Toyoshima I, Yamamoto A, Masamune O, Satake M (1989) Phosphorylation of neurofilament proteins and localization of axonal swellings in motor neuron disease. J Neurol Sci 89:269–277
Ulrich J, Haugh M, Anderton BH, Probst A, Lautenschlager C, His B (1987) Alzheimer dementia and Pick's disease: neurofibrillary tangles and Pick bodies are associated with identical phosphorylated neurofilament epitopes. Acta Neuropathol 73:240–246
Willard M, Simon C (1981) Antibody decoration of neurofilaments. J Cell Biol 89:198–205
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Sloan, K.E., Stevenson, J.A. & Bigbee, J.W. Qualitative and quantitative comparison of the distribution of phosphorylated and non-phosphorylated neurofilament epitopes within central and peripheral axons of adult hamster (Mesocricetus auratus). Cell Tissue Res 263, 265–270 (1991). https://doi.org/10.1007/BF00318768
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DOI: https://doi.org/10.1007/BF00318768