Data on the effect of knockout of cytohesin-1 in myelination-related protein kinase signaling

Cytohesin-1 is the guanine-nucleotide exchange factor of Arf6, a small GTPase of Arf family, and participates in cellular morphological changes. Knockout mice of cytohesin-1 exhibit decreased myelination of neuronal axons in the peripheral nervous system (PNS) “Phosphorylation of cytohesin-1 by Fyn is required for initiation of myelination and the extent of myelination during development (Yamauchi et al., 2012) [1]”. Herein we provide the data regarding decreased phosphorylation levels of protein kinases involved in two major myelination-related kinase cascades in cytohesin-1 knockout mice.


a b s t r a c t
Cytohesin-1 is the guanine-nucleotide exchange factor of Arf6, a small GTPase of Arf family, and participates in cellular morphological changes. Knockout mice of cytohesin-1 exhibit decreased myelination of neuronal axons in the peripheral nervous system (PNS) "Phosphorylation of cytohesin-1 by Fyn is required for initiation of myelination and the extent of myelination during development (Yamauchi et al., 2012) [1]". Herein we provide the data regarding decreased phosphorylation levels of protein kinases involved in two major myelination-related kinase cascades in cytohesin- 1

Value of the data
The data set is of value to the scientific community to need the information for signaling molecules controlling myelination.
The data can provide data for common intracellular signaling cascades involved in myelination.
The data can promote further research on signaling molecules controlling myelination in vivo.

Cytohesin-1 knockout mice
A 13.5-kb Xba I fragment of genomic DNA containing exons 4 to 11 of cytohesin-1 was obtained from a 129/Sv mouse genomic library. The cytohesin-1-targeting vector was constructed by replacing thẽ 3.6-kb Xba I fragment containing exons 4 to 7 of cytohesin-1 within the fragment containing exons 4 to 11, which was ligated to the gene encoding diphtheria toxin, with a cassette of the neomycinresistant gene. 129/Sv embryonic stem (ES) cells were transfected with the linearized targeting vector by electroporation. These ES cells were used to generate chimeric mice. Heterozygous offspring were mated to wild-type C57BL/6JJms mice, and the mutations were propagated in this strain for at least 10 generations before it was crossed to produce homozygotes for experiments. Homozygous mice, as well as heterozygous mice, were fertile under standard breeding conditions [1]. The genomic PCR for identification of the knockout allele was performed. The primers used for genomic PCR were 5'-CCCGGTTCTTTTTGTCAAGACCGACCTGTC-3' (sense) and 5'-CATTCGCCGCCAAGCTCTTCAGCAATATCAC-3' (antisense) for the neo gene [1]. PCR amplification was performed in 30 cycles, each consisting of denaturation at 94°C for 1 min, annealing at 68°C for 1 min, and extension at 72°C for 1 min. Male mice were used for experiments if it was possible to distinguish their sex.

Statistical analysis
Data are presented as means 7 S.D. from independent experiments. Intergroup comparisons were performed using unpaired Student's t-test. Differences were considered significant when p value was less than 0.01.