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Phosphorylation of GAP-43 T172 is a molecular marker of growing axons in a wide range of mammals including primates

GAP-43 is a vertebrate neuron-specific protein and that is strongly related to axon growth and regeneration; thus, this protein has been utilized as a classical molecular marker of these events and growth cones. Although GAP-43 was biochemically characterized more than a quarter century ago, how thi...

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Autores principales: Okada, Masayasu, Kawagoe, Yosuke, Sato, Yuta, Nozumi, Motohiro, Ishikawa, Yuya, Tamada, Atsushi, Yamazaki, Hiroyuki, Sekino, Yuko, Kanemura, Yonehiro, Shinmyo, Yohei, Kawasaki, Hiroshi, Kaneko, Naoko, Sawamoto, Kazunobu, Fujii, Yukihiko, Igarashi, Michihiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8034164/
https://www.ncbi.nlm.nih.gov/pubmed/33832520
http://dx.doi.org/10.1186/s13041-021-00755-0
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author Okada, Masayasu
Kawagoe, Yosuke
Sato, Yuta
Nozumi, Motohiro
Ishikawa, Yuya
Tamada, Atsushi
Yamazaki, Hiroyuki
Sekino, Yuko
Kanemura, Yonehiro
Shinmyo, Yohei
Kawasaki, Hiroshi
Kaneko, Naoko
Sawamoto, Kazunobu
Fujii, Yukihiko
Igarashi, Michihiro
author_facet Okada, Masayasu
Kawagoe, Yosuke
Sato, Yuta
Nozumi, Motohiro
Ishikawa, Yuya
Tamada, Atsushi
Yamazaki, Hiroyuki
Sekino, Yuko
Kanemura, Yonehiro
Shinmyo, Yohei
Kawasaki, Hiroshi
Kaneko, Naoko
Sawamoto, Kazunobu
Fujii, Yukihiko
Igarashi, Michihiro
author_sort Okada, Masayasu
collection PubMed
description GAP-43 is a vertebrate neuron-specific protein and that is strongly related to axon growth and regeneration; thus, this protein has been utilized as a classical molecular marker of these events and growth cones. Although GAP-43 was biochemically characterized more than a quarter century ago, how this protein is related to these events is still not clear. Recently, we identified many phosphorylation sites in the growth cone membrane proteins of rodent brains. Two phosphorylation sites of GAP-43, S96 and T172, were found within the top 10 hit sites among all proteins. S96 has already been characterized (Kawasaki et al., 2018), and here, phosphorylation of T172 was characterized. In vitro (cultured neurons) and in vivo, an antibody specific to phosphorylated T172 (pT172 antibody) specifically recognized cultured growth cones and growing axons in developing mouse neurons, respectively. Immunoblotting showed that pT172 antigens were more rapidly downregulated throughout development than those of pS96 antibody. From the primary structure, this phosphorylation site was predicted to be conserved in a wide range of animals including primates. In the developing marmoset brainstem and in differentiated neurons derived from human induced pluripotent stem cells, immunoreactivity with pT172 antibody revealed patterns similar to those in mice. pT172 antibody also labeled regenerating axons following sciatic nerve injury. Taken together, the T172 residue is widely conserved in a wide range of mammals including primates, and pT172 is a new candidate molecular marker for growing axons.
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spelling pubmed-80341642021-04-12 Phosphorylation of GAP-43 T172 is a molecular marker of growing axons in a wide range of mammals including primates Okada, Masayasu Kawagoe, Yosuke Sato, Yuta Nozumi, Motohiro Ishikawa, Yuya Tamada, Atsushi Yamazaki, Hiroyuki Sekino, Yuko Kanemura, Yonehiro Shinmyo, Yohei Kawasaki, Hiroshi Kaneko, Naoko Sawamoto, Kazunobu Fujii, Yukihiko Igarashi, Michihiro Mol Brain Research GAP-43 is a vertebrate neuron-specific protein and that is strongly related to axon growth and regeneration; thus, this protein has been utilized as a classical molecular marker of these events and growth cones. Although GAP-43 was biochemically characterized more than a quarter century ago, how this protein is related to these events is still not clear. Recently, we identified many phosphorylation sites in the growth cone membrane proteins of rodent brains. Two phosphorylation sites of GAP-43, S96 and T172, were found within the top 10 hit sites among all proteins. S96 has already been characterized (Kawasaki et al., 2018), and here, phosphorylation of T172 was characterized. In vitro (cultured neurons) and in vivo, an antibody specific to phosphorylated T172 (pT172 antibody) specifically recognized cultured growth cones and growing axons in developing mouse neurons, respectively. Immunoblotting showed that pT172 antigens were more rapidly downregulated throughout development than those of pS96 antibody. From the primary structure, this phosphorylation site was predicted to be conserved in a wide range of animals including primates. In the developing marmoset brainstem and in differentiated neurons derived from human induced pluripotent stem cells, immunoreactivity with pT172 antibody revealed patterns similar to those in mice. pT172 antibody also labeled regenerating axons following sciatic nerve injury. Taken together, the T172 residue is widely conserved in a wide range of mammals including primates, and pT172 is a new candidate molecular marker for growing axons. BioMed Central 2021-04-08 /pmc/articles/PMC8034164/ /pubmed/33832520 http://dx.doi.org/10.1186/s13041-021-00755-0 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Okada, Masayasu
Kawagoe, Yosuke
Sato, Yuta
Nozumi, Motohiro
Ishikawa, Yuya
Tamada, Atsushi
Yamazaki, Hiroyuki
Sekino, Yuko
Kanemura, Yonehiro
Shinmyo, Yohei
Kawasaki, Hiroshi
Kaneko, Naoko
Sawamoto, Kazunobu
Fujii, Yukihiko
Igarashi, Michihiro
Phosphorylation of GAP-43 T172 is a molecular marker of growing axons in a wide range of mammals including primates
title Phosphorylation of GAP-43 T172 is a molecular marker of growing axons in a wide range of mammals including primates
title_full Phosphorylation of GAP-43 T172 is a molecular marker of growing axons in a wide range of mammals including primates
title_fullStr Phosphorylation of GAP-43 T172 is a molecular marker of growing axons in a wide range of mammals including primates
title_full_unstemmed Phosphorylation of GAP-43 T172 is a molecular marker of growing axons in a wide range of mammals including primates
title_short Phosphorylation of GAP-43 T172 is a molecular marker of growing axons in a wide range of mammals including primates
title_sort phosphorylation of gap-43 t172 is a molecular marker of growing axons in a wide range of mammals including primates
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8034164/
https://www.ncbi.nlm.nih.gov/pubmed/33832520
http://dx.doi.org/10.1186/s13041-021-00755-0
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