Abstract
In contrast to peripheral nerves, central neurons do not regrow spontaneously after injury. Our previous studies showed that transplantation of degenerating peripheral nerves or their extracts can induce regeneration in the injured central nervous system. Non-predegenerated nerves show much weaker neurotrophic activity. The aim of the present work was to examine quantitatively and qualitatively the protein composition of rat sciatic nerve extracts. The experiments were carried out on male Wistar C rats. Distal fragments were collected immediately after transection or after 7 day-long predegeneration. The nerves were homogenized, centrifuged and ultracetrifuged. Extracts were analyzed by means of two-dimensional electrophoresis. The two-dimensional electrophoresis showed 69 protein subfractions with isoelectric points ranging from 4.2 to 7.0 pH and molecular weight ranging from 13.5 kDa to 335.4 kDa in extracts obtained from nonpredegenerated nerves. In predegenerated nerve extracts 114 subfractions with isoelectric points ranging from 4.2 to 7.4 pH and molecular weight from 21.1 kDa to 335.4 kDa were found. Fractions: 25.5 kDa, 31.6 kDa, 36 kDa, 38.4 kDa, 42.4 kDa, 46.6 kDa, and 50.5 kDa showed significant increase and two fractions: 68.5 kDa and 335.4 kDa demonstrated significant decrease in the number of subfractions in predegenerated nerves. Fractions 160.8 kDa, 236.1 kDa, and 5 fractions below 21.1 kDa were present only in extracts from non-predegenerated nerves. In conclusion, the results of our study demonstrate that the most intense changes in protein composition in degenerating nerves take place in low molecular weight fractions.
Highlights
It is proven that the damaged neurons of the central nervous system (CNS) of adult mammals, in contrast to peripheral nervous system, do not have the capacity for spontaneous and successful regrowth
It is possible to intensify their neurotrophic effect by extraction proteins of molecular weight from 10 to 100 kDa from the entire extract [in press – Neurotrophic activity of extracts from distal stumps of predegenerated peripheral rat nerves varies according to molecular mass spectrum[5]
The two-dimensional electrophoresis showed 28 protein fractions of different molecular weights containing 69 protein subfractions with pI ranging from 4.2 to 7.0 in extracts obtained from nonpredegenerated nerves
Summary
It is proven that the damaged neurons of the central nervous system (CNS) of adult mammals, in contrast to peripheral nervous system, do not have the capacity for spontaneous and successful regrowth. It is possible to intensify their neurotrophic effect by extraction proteins of molecular weight from 10 to 100 kDa from the entire extract [in press – Neurotrophic activity of extracts from distal stumps of predegenerated peripheral rat nerves varies according to molecular mass spectrum[5]. This forms the rationale for selecting these extracts for further investigation
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