Synuclein Analysis in Adult <i>Xenopus laevis</i>
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 <i>Torpedo californica</i>, synucleins have been sequenced in all vertebrates, showing differences...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-05-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/11/6058 |
| _version_ | 1797493162348904448 |
|---|---|
| author | Maria Carmela Bonaccorsi di Patti Elisa Angiulli Arianna Casini Rosa Vaccaro Carla Cioni Mattia Toni |
| author_facet | Maria Carmela Bonaccorsi di Patti Elisa Angiulli Arianna Casini Rosa Vaccaro Carla Cioni Mattia Toni |
| author_sort | Maria Carmela Bonaccorsi di Patti |
| collection | DOAJ |
| description | 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 <i>Torpedo californica</i>, 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 <i>Xenopus laevis</i> 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 <i>Xenopus</i> synucleins was evaluated by circular dichroism analysis. Results indicate <i>Xenopus</i> as a good model for studying synucleinopathies, and provide a useful background for future studies on synuclein functions and their evolution in vertebrates. |
| first_indexed | 2024-03-10T01:16:03Z |
| format | Article |
| id | doaj.art-5a68adb6cdf24b81aa7a851993939dde |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T01:16:03Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-5a68adb6cdf24b81aa7a851993939dde2023-11-23T14:08:52ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-012311605810.3390/ijms23116058Synuclein Analysis in Adult <i>Xenopus laevis</i>Maria Carmela Bonaccorsi di Patti0Elisa Angiulli1Arianna Casini2Rosa Vaccaro3Carla Cioni4Mattia Toni5Department of Biochemical Sciences, Sapienza University, 00185 Rome, ItalyDepartment of Biology and Biotechnology “Charles Darwin”, Sapienza University, 00161 Rome, ItalyDepartment of Anatomical, Histological, Forensic Medicine and Orthopedic Sciences, Sapienza University, 00161 Rome, ItalyDepartment of Anatomical, Histological, Forensic Medicine and Orthopedic Sciences, Sapienza University, 00161 Rome, ItalyDepartment of Biology and Biotechnology “Charles Darwin”, Sapienza University, 00161 Rome, ItalyDepartment of Biology and Biotechnology “Charles Darwin”, Sapienza University, 00161 Rome, ItalyThe 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 <i>Torpedo californica</i>, 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 <i>Xenopus laevis</i> 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 <i>Xenopus</i> synucleins was evaluated by circular dichroism analysis. Results indicate <i>Xenopus</i> as a good model for studying synucleinopathies, and provide a useful background for future studies on synuclein functions and their evolution in vertebrates.https://www.mdpi.com/1422-0067/23/11/6058synuclein<i>Xenopus laevis</i>Western blotrecombinant proteinsqRT-PCR |
| spellingShingle | Maria Carmela Bonaccorsi di Patti Elisa Angiulli Arianna Casini Rosa Vaccaro Carla Cioni Mattia Toni Synuclein Analysis in Adult <i>Xenopus laevis</i> International Journal of Molecular Sciences synuclein <i>Xenopus laevis</i> Western blot recombinant proteins qRT-PCR |
| title | Synuclein Analysis in Adult <i>Xenopus laevis</i> |
| title_full | Synuclein Analysis in Adult <i>Xenopus laevis</i> |
| title_fullStr | Synuclein Analysis in Adult <i>Xenopus laevis</i> |
| title_full_unstemmed | Synuclein Analysis in Adult <i>Xenopus laevis</i> |
| title_short | Synuclein Analysis in Adult <i>Xenopus laevis</i> |
| title_sort | synuclein analysis in adult i xenopus laevis i |
| topic | synuclein <i>Xenopus laevis</i> Western blot recombinant proteins qRT-PCR |
| url | https://www.mdpi.com/1422-0067/23/11/6058 |
| work_keys_str_mv | AT mariacarmelabonaccorsidipatti synucleinanalysisinadultixenopuslaevisi AT elisaangiulli synucleinanalysisinadultixenopuslaevisi AT ariannacasini synucleinanalysisinadultixenopuslaevisi AT rosavaccaro synucleinanalysisinadultixenopuslaevisi AT carlacioni synucleinanalysisinadultixenopuslaevisi AT mattiatoni synucleinanalysisinadultixenopuslaevisi |