Synuclein Analysis in Adult Xenopus laevis

• Published in: International journal of molecular sciences 2022-05, Vol.23 (11), p.6058
• Main Authors: Bonaccorsi di Patti, Maria Carmela, Angiulli, Elisa, Casini, Arianna, Vaccaro, Rosa, Cioni, Carla, Toni, Mattia
• Format: Article
• Language: English
• Subjects: Alternative splicing; Amino acids; Amyloidogenesis; Antibodies; Binding sites; Brain research; Chromosomes; Circular dichroism; Dementia; Dichroism; Evolutionary conservation; Gene expression; Genomes; Homology; Immunohistochemistry; Inclusions; Isoforms; Mammals; Mutation; Nervous system; Neuronal-glial interactions; Organs; Parkinson's disease; Physiology; Protein expression; Protein structure; Proteins; qRT-PCR; recombinant proteins; Secondary structure; Spinal cord; Spleen; Splicing; Synuclein; Vertebrates; Western blot; Xenopus; Xenopus laevis
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23116058

The α-, β- and γ-synucleins are small soluble proteins expressed in the nervous system of mammals and evolutionary conserved in vertebrates. After being discovered in the cartilaginous fish , synucleins have been sequenced in all vertebrates, showing differences in the number of genes and splicing isoforms in different taxa. Although α-, β- and γ-synucleins share high homology in the N-terminal sequence, suggesting their evolution from a common ancestor, the three isoforms also differ in molecular characteristics, expression levels and tissue distribution. Moreover, their functions have yet to be fully understood. Great scientific interest on synucleins mainly derives from the involvement of α-synuclein in human neurodegenerative diseases, collectively named synucleinopathies, which involve the accumulation of amyloidogenic α-synuclein inclusions in neurons and glia cells. Studies on synucleinopathies can take advantage of the development of new vertebrate models other than mammals. Moreover, synuclein expression in non-mammalian vertebrates contribute to clarify the physiological role of these proteins in the evolutionary perspective. In this paper, gene expression levels of α-, β- and γ-synucleins have been analysed in the main organs of adult by qRT-PCR. Moreover, recombinant α-, β- and γ-synucleins were produced to test the specificity of commercial antibodies against α-synuclein used in Western blot and immunohistochemistry. Finally, the secondary structure of synucleins was evaluated by circular dichroism analysis. Results indicate as a good model for studying synucleinopathies, and provide a useful background for future studies on synuclein functions and their evolution in vertebrates.