| α-Synuclein (α-Syn), an acidic protein with 140 amino acid residues, expressed highly in the brain, and mainly clustered in the neurons presynaptic endings.α-Syn plays important roles in development and formation of neuronal, dopamine synthesis, maintance of synaptic plasticity and transportation of synaptic vesicle. Much work onα-Syn has been done in human and other mammal species, while little in pigeon. Nucleotide sequence ofα-Syn and its distribution in the brain of king pigeon has not been reported.In this study,α-Syn gene coding region was cloned by molecular biology technology;α-Syn cDNA coding region nucleotide and amino acid sequence alignment were conducted between king pigeon and other 7 species such as Gullus gullus, taeniopygia guttata, Pantroglodytes, Homo sapiens, Mus musculus, Sus scrofa, Bos Taurus and the biological evolutionary tree was built;α-Syn was expressed in prokaryotic system by genetic engineering technology, and the polyclonal antibody was prepared using expressed fusion protein; The distribution ofα-Syn in the brain was detected by immunohistochemistry; AVM acute poisoning model was built and its effect on the protein expression ofα-Syn was detected. The main results were showed as follows:1 The results of sequence analysis showed that there was a 613 bp cDNA sequence ofα-Syn gene cloned, including 432 bp ORF with start codon ATG and stop codon TGA, encoding 143 amino acide, 16 bp 5′UTR and 165 bp 3′UTR. The protein molecular weight is 14.9 kDa.2 The homology ofα-Syn coding region between king pigeon and the other 7 species is 81.71%~93.52%, and the highest homology is 93.52% between king pigeon and both Gullus gullus and taeniopygia guttata. Judging from the amino acid sequences comparison, the homology between king pigeon and the other 7 species is 83.22%~97.20%.α-Syn genes of different species was highly conservative, which suggested that the function ofα-Syn in king pigeon and other known species genes may be similar.3 Fusion proteins of pGEX6p-1-α-Syn were expressed in E.coli expression system. The polyclonal antibody was prepared by using expressed fusion proteins. The antibody titer was 1:51 200 and polyclonal antibody is specificity for fusion protein detected by Western blot.4α-Syn was detected in all 5 tissues incluing hippocampus, thalamus, optic lobe, epencephalon and olfactory bulb in king pigeon, and the order ofα-Syn contents from high to low was hippocampus, optic lobe, thalamus, olfactory bulb and epencephalon.5 AVM acute poisoning increased the protein expression ofα-Syn in all 5 tissues. The differences ofα-Syn expression in hippocampus, optic lobe and thalamus were significant (P<0.05), while there were no significant differences in the olfactory bulb and epencephalon between AVM acute poisoning group and the control group (P>0.05). These rusults indicated that increase ofα-Syn expressions in different brain tissues was different, which suggested that nerve injury induced by AVM acute poisoning was associated with increase ofα-Syn expression. |
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