| Objective:By using the G93A-SOD1 transgenic mice as the animal models of amyotrophic lateral sclerosis(ALS) to initially proved the relationship between disease-causing protein(FUS/TLS)in adult spinal cord at the different stages of ALS and motor neuron progressive death, to find the abnormal expression and distribution of the neurons disease-causing protein in the spinal cord of the ALS mice. It?s possible to find a way to the pathogenesis of ALS. Methods:Use the SOD1 G93 A C57BL/6J transgenic mice as the animal models which be detected by PCR and select the transgenic mice which we want. The SOD1 G93 A transgenic animals fixed with formalin, then took out the mouse spinal cord and brain compeletly, and the sample was dehydrated, embedded, frozen section in order. Use the dual fluorescence immunofluorescence staining to label the Fused in Sarcoma/Translocated in Liposarcoma Protein(FUS/TLS), neurons, astrocytes, oligodendrocytes. Every anatomical region was stained double immunofluorescence label.The cells were observed in the same vision and the same magnification of the fluorescencemicroscope, the two different stained positive cells were used the image processing technology to overlay. Finally, observe the distribution and number of the FUS/TLS positive cells in the different anatomical regions and stages, and analyze the relationship with the neuron. Results:1. The FUS/TLS almost didn?t expressed in adult spinal cord in the SOD1 wild-type transgenic mice and the pre-onset stages of SOD1 G93 A transgenic mice, only expressed in adult spinal cord at the onset and progression stages of the SOD1 G93 A transgenic mice.2. The increase of FUS/TLS expression was significantly different in the different anatomic regions of spinal cord at the onset and progression stages of ALS-like mice. The increase rank order is thoracic>cervical>lumbar segment, the anterior funiculus>lateral funiculus>the gray matter of ventral horn>the gray matter surrounding the central canal>the gray matter of dorsal horn>posterior funiculus.3. The abnormal expression of FUS/TLS only occurred in the astrocyte cell, wasn?t observed the abnormal expression in the other neural cells including neuron and oligodendrocyte cell.4. The FUS/TLS only expressed in the nuclear of astrocyte cells at the onset stages of the SOD1 G93 A transgenic mice. almost all FUS/TLS expressed in the cytoplasm of astrocyte cells at the progression stages,little in the nuclear.5. The increase of FUS/TLS expression showed a negatively relationship with the amount of neuron cells in spinal cord at the onset and progression stages of ALS mice. Conclusions:The abnormal expression and mislocation of FUS/TLS in the astrocyte cell causes progressive motor neuron degeneration in the ALS-like SOD1 G93 A transgenic mice. |
