Characterization of pip5k3 fleck corneal dystrophy-linked gene in zebrafish.
PIKfyve is a kinase encoded by pip5k3 involved in phosphatidylinositols (PdtIns) pathways. These lipids building cell membranes have structural functions and are involved in complex intracellular regulations. Mutations in human PIP5K3 are associated with François-Neetens mouchetée fleck corneal dystrophy [Li, S., Tiab, L., Jiao, X., Munier, F.L., Zografos, L., Frueh, B.E., Sergeev, Y., Smith, J., Rubin, B., Meallet, M.A., Forster, R.K., Hejtmancik, J.F., Schorderet, D.F., 2005. Mutations in PIP5K3 are associated with François-Neetens mouchetee fleck corneal dystrophy. Am. J. Hum. Genet. 77, 54-63]. We cloned the zebrafish pip5k3 and report its molecular characterization and expression pattern in adult fish as well as during development. The zebrafish PIKfyve was 70% similar to the human homologue. The gene encompassed 42 exons and presented four alternatively spliced variants. It had a widespread expression in the adult organs and was localized in specific cell types in the eye as the cornea, lens, ganglion cell layer, inner nuclear layer and outer limiting membrane. Pip5k3 transcripts were detected in early cleavage stage embryos. Then it was uniformly expressed at 10 somites, 18 somites and 24 hpf. Its expression was then restricted to the head region at 48 hpf, 72 hpf and 5 dpf and partial expression was found in somites at 72 hpf and 5 dpf. In situ on eye sections at 3 dpf showed a staining mainly in lens, outer limiting membrane, inner nuclear layer and ganglion cell layer. A similar expression pattern was found in the eye at 5 dpf. A temporal regulation of the spliced variants was observed at 1, 3 and 5 dpf and they were also found in the adult eye.
IRO, Institute for Research in Ophthalmology, Grand-Champsec 64, 1950 Sion, Switzerland; University of Lausanne, Department of Ophthalmology, 1015 Lausanne, Switzerland.
Gaëlle
G
Boisset
IRO, Institute for Research in Ophthalmology, Grand-Champsec 64, 1950 Sion, Switzerland.
Bożena K
BK
Polok
IRO, Institute for Research in Ophthalmology, Grand-Champsec 64, 1950 Sion, Switzerland; University of Lausanne, Department of Ophthalmology, 1015 Lausanne, Switzerland; Swiss Federal Institute of Technology, 1015 Lausanne, Switzerland.
Daniel F
DF
Schorderet
D017398:Alternative Splicing; D000818:Animals; D003001:Cloning, Molecular; D004625:Embryo, Nonmammalian / Q000201:enzymology; D005123:Eye / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D014018:Tissue Distribution; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000378:metabolism; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
D012333:RNA, Messenger; D029961:Zebrafish Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C532057:pikfyve protein, zebrafish