Upregulation of leukemia inhibitory factor (LIF) during the early stage of optic nerve regeneration in zebrafish

Kazuhiro Ogai; Ayaka Kuwana; Suguru Hisano; Mikiko Nagashima; Yoshiki Koriyama; Kayo Sugitani; Kazuhiro Mawatari; Hiroshi Nakashima; Satoru Kato

doi:10.1371/journal.pone.0106010

Upregulation of leukemia inhibitory factor (LIF) during the early stage of optic nerve regeneration in zebrafish

PLoS One. 2014 Aug 27;9(8):e106010. doi: 10.1371/journal.pone.0106010. eCollection 2014.

Authors

Kazuhiro Ogai¹, Ayaka Kuwana², Suguru Hisano², Mikiko Nagashima³, Yoshiki Koriyama⁴, Kayo Sugitani², Kazuhiro Mawatari², Hiroshi Nakashima², Satoru Kato³

Affiliations

¹ Department of Molecular Neurobiology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa, Japan; Wellness Promotion Science Center, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan.
² Department of Clinical Laboratory Science, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa, Japan.
³ Department of Molecular Neurobiology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa, Japan.
⁴ Department of Molecular Neurobiology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa, Japan; Graduate School and Faculty of Pharmaceutical Sciences, Suzuka University of Medical Science, Suzuka, Mie, Japan.

Abstract

Fish retinal ganglion cells (RGCs) can regenerate their axons after optic nerve injury, whereas mammalian RGCs normally fail to do so. Interleukin 6 (IL-6)-type cytokines are involved in cell differentiation, proliferation, survival, and axon regrowth; thus, they may play a role in the regeneration of zebrafish RGCs after injury. In this study, we assessed the expression of IL-6-type cytokines and found that one of them, leukemia inhibitory factor (LIF), is upregulated in zebrafish RGCs at 3 days post-injury (dpi). We then demonstrated the activation of signal transducer and activator of transcription 3 (STAT3), a downstream target of LIF, at 3-5 dpi. To determine the function of LIF, we performed a LIF knockdown experiment using LIF-specific antisense morpholino oligonucleotides (LIF MOs). LIF MOs, which were introduced into zebrafish RGCs via a severed optic nerve, reduced the expression of LIF and abrogated the activation of STAT3 in RGCs after injury. These results suggest that upregulated LIF drives Janus kinase (Jak)/STAT3 signaling in zebrafish RGCs after nerve injury. In addition, the LIF knockdown impaired axon sprouting in retinal explant culture in vitro; reduced the expression of a regeneration-associated molecule, growth-associated protein 43 (GAP-43); and delayed functional recovery after optic nerve injury in vivo. In this study, we comprehensively demonstrate the beneficial role of LIF in optic nerve regeneration and functional recovery in adult zebrafish.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Diffusion
GAP-43 Protein / genetics
GAP-43 Protein / metabolism
Gene Expression Regulation
Janus Kinase 1 / genetics
Janus Kinase 1 / metabolism
Leukemia Inhibitory Factor / antagonists & inhibitors
Leukemia Inhibitory Factor / genetics*
Leukemia Inhibitory Factor / metabolism
Morpholinos / genetics
Morpholinos / metabolism
Nerve Regeneration / genetics*
Oligonucleotides, Antisense / genetics
Oligonucleotides, Antisense / metabolism
Optic Nerve / metabolism
Optic Nerve / pathology
Optic Nerve Injuries / genetics*
Optic Nerve Injuries / metabolism
Optic Nerve Injuries / pathology
Optic Nerve Injuries / rehabilitation
Recovery of Function / physiology
Retinal Ganglion Cells / metabolism*
Retinal Ganglion Cells / pathology
STAT3 Transcription Factor / genetics*
STAT3 Transcription Factor / metabolism
Signal Transduction
Time Factors
Tissue Culture Techniques
Zebrafish
Zebrafish Proteins / antagonists & inhibitors
Zebrafish Proteins / genetics*
Zebrafish Proteins / metabolism

Substances

GAP-43 Protein
Leukemia Inhibitory Factor
Morpholinos
Oligonucleotides, Antisense
STAT3 Transcription Factor
Zebrafish Proteins
m17 protein, zebrafish
stat3 protein, zebrafish
Janus Kinase 1

Grants and funding

This work was supported in part by the Japan Society for the Promotion of Science KAKENHI Grant Numbers 25890007 (KO), 23618006 (KS), 25462753 (YK), 23650163 and 25640008 (SK). This study was also funded in part by the Ministry of Education, Culture, Sports, Science and Technology/Japan Science and Technology Agency Tenure Track Promotion Program (KO). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.