A Two-Dimensional Gel Electrophoretic Study of Proteins Synthesized and Released by Degenerating Adult Mouse Sciatic Nerve

• Published in: Experimental neurology 2000-03, Vol.162 (1), p.194-200
• Main Authors: Golding, Jon P., Tonge, David
• Format: Article
• Language: English
• Subjects: 2-D gel electrophoresis; Age Factors; Animals; Biological and medical sciences; Cells, Cultured; degeneration; Densitometry; Electrophoresis, Gel, Two-Dimensional; Female; Freezing; Injuries of the nervous system and the skull. Diseases due to physical agents; Isoelectric Point; Macrophages; Medical sciences; Mice; Mice, Inbred Strains; Molecular Weight; Nerve Degeneration - metabolism; Nerve Tissue Proteins - biosynthesis; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - secretion; Neurons - cytology; Neurons - metabolism; Neurons - secretion; PNS; protein secretion; protein synthesis; regeneration; Schwann Cells; Sciatic Nerve - metabolism; Sciatic Nerve - pathology; Traumas. Diseases due to physical agents
• ISSN: 0014-4886; 1090-2430
• DOI: 10.1006/exnr.1999.7332

Previous two-dimensional (2-D) gel electrophoretic studies of proteins secreted by degenerating mammalian peripheral nerves (Ignatius et al., 1986, Proc. Natl. Acad. Sci. USA 83: 1125–1129; Muller et al., 1986, J. Cell Biol. 102: 393–402) detected the up-regulation of two proteins of 67–70 and 34–37 kDa, although they failed to resolve proteins smaller than about 15 kDa or with isoelectric points greater than 8. In the present study, we have used 2-D gels that can resolve proteins in the molecular mass range 3.6–200 kDa and isoelectric point range 2.4–10.6. This revealed that the incorporation of radiolabel by three diffusible proteins with apparent molecular mass/isoelectric point values of 38/5–6, 27–31/4–5, and 8/>10 was increased in the distal stumps of sciatic nerves 4 days after lesion, while the radiolabel incorporation by a further two proteins (15/5.3 and 12.5–17.5/6.8–7.5) was increased in the distal nerve stump 15 days after lesion. The possible cellular sources of these proteins were assessed by comparing protein secretion from unoperated nerves with nerve segments maintained in culture for 4 days (in which the contribution from recruited macrophages would be expected to be minimal) and segments of nerve that had been frozen and then replaced in situ for 4 days (in which the contribution from nerve sheath cells would be expected to be minimal). This revealed that three of the proteins up-regulated in lesioned nerves (27–31/4–5, 15/5.3, and 12.5–17.5/6.8–7.5) are probably sheath cell products, while the other two (38/5–6 and 8/>10) may be secreted mainly by macrophages (or other cells) that infiltrate the frozen nerve segments. The identity of these proteins and their possible involvement in axonal regeneration remain to be determined.