Van den Haute C, Spittaels K, Van Dorpe J, Lasrado R, Vandezande K, Laenen I, Geerts H, Van Leuven F.
Coexpression of human cdk5 and its activator p35 with human protein tau in neurons in brain of triple transgenic mice.Neurobiol Dis.
2001 Feb;8(1):32-44.
PubMed Abstract
REAGENTS/MATERIAL: Human cdk5 cDNA (gift from I. Hoffmann, Heidelberg, Germany)
p35 cDNA (gift from L.H. Tsai, Boston, MA)
Human tau40 trasgenic mice were generated as described (Spittaels et al)
cdk5 PCR probe: forward 5'-CCCCACCACAGAATCCA-3'
reverse 5'-TAACAGCGGACGGGAATC-3'
p35 PCR probe: forward 5'-CCCCACCACAGAATCCA-3'
reverse 5'-CAAGGTCCCCGTTTCTCC-3'
tau40 PCR probe:forward 5'-ACCCCATCCCTACCAACA-3'
reverse 5'-GCAGGCGGCTCTTACTAG-3'
cdk5 polyclonal antibody (H-291 at 67 ng/ml) (Santa Cruz Biotechnology)
p35 polyclonal antibody (N-20 at 5 ug/ml) (Santa Cruz Biotechnology)
p35 polyclonal antibody (C-19 at 400 ng/ml) (Santa Cruz Biotechnology)
Tau-1 MAb (1 ug/ml) (Boeringer Mannheim)
TAU-5 MAb (500 ng/ml) (Pharmingen)
AT-8 MAb (2 ug/ml) (Mercken et al)
AT-180 (1 ug/ml) (Innogenetics)
AD-2 (1 ug/ml) (gift from B. Pau, Lille, France)
PHF-1 (1/200) (gift from P. Davies, New York, NY)
NF-200 MAb (Sigma) is phosphorylation-independent
SMI-31, and SMI-32 MAbs (Sternberger) are phosphorylation-dependent
FUTURE DIRECTION: Results show that cdk5/p35 did not cause an appreciable hyperphosphorylation of murine or human protein tau in vivo. Further study on the relationship of the cdk5 kinase activity to protein tau is needed.