Expression of neurofilament gene in spinal motoneurons during neural regeneration

The techniquein situ hybridization was used to measure the levels of light (NF-L), medium (NF-M) and heavy (NF-H) neurofilament protein subunits mRNA in L_4-6 spinal motoneurons in adult rat during regeneration following a unilateral crush of the sciatic nerve. It was found that the hybridization signals of each neurofilament subunit rnRNA were dramatically decreased in spinal motoneurons postaxotomy by light microscopy. The hybridization signals of NF-L and NF-M mRNA were located in cytoplasm of neurons, whereas NF-H mRNA was found in both nucleus and cytoplasm of neurons. Image analysis showed that the encoding levels of mRNA for each of neurofilament subunit mRNA reduced on the 3rd d and returned to control levels on the 28th d following the lesion. The relative levels of mRNA coding for each neurofilament subunit were significantly different. The lowest level of NF-L mRNA was observed at 5 d postaxotomy, and that of NF-M, NF-H mRNA on the 7th and 10th d after injury. Moreover, the levels of HF-M and NF-H mRNA were reduced much lower and lasted much longer than that of NF-L mRNA. The observations suggest that there were different mechanisms for the regulation of neurofilament subunit genes expression. The reduced neurofilament gene expression may be due to a response to axonal injury and advance the restructure of axonal architecture.