Modulation of p53 and met expression by Krüppel‐like factor 8 regulates zebrafish cerebellar development

• Published in: Developmental neurobiology (Hoboken, N.J.) N.J.), 2015-09, Vol.75 (9), p.908-926
• Main Authors: Tsai, Ming‐Yuan, Lu, Yu‐Fen, Liu, Yu‐Hsiu, Lien, Huang‐Wei, Huang, Chang‐Jen, Wu, Jen‐Leih, Hwang, Sheng‐Ping L.
• Format: Article
• Language: English
• Subjects: Animals; Animals, Genetically Modified; apoptosis; Apoptosis - physiology; cerebellum; Cerebellum - embryology; Cerebellum - metabolism; Cerebellum - pathology; Gene Knockdown Techniques; Klf8; Kruppel-Like Transcription Factors - genetics; Kruppel-Like Transcription Factors - metabolism; met; Models, Animal; Morpholinos; Mutation; Neural Stem Cells - physiology; Neurogenesis - physiology; Neurons - metabolism; Neurons - pathology; p53; Proto-Oncogene Proteins c-met - metabolism; RNA, Messenger - metabolism; Tectum Mesencephali - embryology; Tectum Mesencephali - metabolism; Tectum Mesencephali - pathology; Transcription Factors - metabolism; Tumor Suppressor Protein p53 - metabolism; Zebrafish; Zebrafish Proteins - genetics; Zebrafish Proteins - metabolism
• ISSN: 1932-8451; 1932-846X
• DOI: 10.1002/dneu.22258

ABSTRACT Krüppel‐like factor 8 (Klf8) is a zinc‐finger transcription factor implicated in cell proliferation, and cancer cell survival and invasion; however, little is known about its role in normal embryonic development. Here, we show that Klf8 is required for normal cerebellar development in zebrafish embryos. Morpholino knockdown of klf8 resulted in abnormal cerebellar primordium morphology and the induction of p53 in the brain region at 24 hours post‐fertilization (hpf). Both p53‐dependent reduction of cell proliferation and augmentation of apoptosis were observed in the cerebellar anlage of 24 hpf‐klf8 morphants. In klf8 morphants, expression of ptf1a in the ventricular zone was decreased from 48 to 72 hpf; on the other hand, expression of atohla in the upper rhombic lip was unaffected. Consistent with this finding, Purkinje cell development was perturbed and granule cell number was reduced in 72 hpf‐klf8 morphants; co‐injection of p53 MOsp or klf8 mRNA substantially rescued development of cerebellar Purkinje cells in klf8 morphants. Hepatocyte growth factor/Met signaling is known to regulate cerebellar development in zebrafish and mouse. We observed decreased met expression in the tectum and rhombomere 1 of 24 hpf‐klf8 morphants, which was largely rescued by co‐injection with klf8 mRNA. Moreover, co‐injection of met mRNA substantially rescued formation of Purkinje cells in klf8 morphants at 72 hpf. Together, these results demonstrate that Klf8 modulates expression of p53 and met to maintain ptf1a‐expressing neuronal progenitors, which are required for the appropriate development of cerebellar Purkinje and granule cells in zebrafish embryos. © 2014 Wiley Periodicals, Inc. Develop Neurobiol 75: 908–926, 2015