Up-regulation of a non-kinase activity isoform of Ca(2+)/calmodulin-dependent protein kinase kinase beta1 (CaMKKbeta1) in hibernating bat brain.

Hibernation is an adaptive strategy that is utilized by some animals to survive the harsh environments of low temperature and food scarce. Hibernators, however, can survive in frequent and dramatic fluctuation of body temperature and blood flow causing by periodic arousals during hibernation without brain insult, and this indicates that it must have some unique adaptive aspects of hibernation physiology. To find out the up-regulated genes of bat brain during hibernation and explore the brain function adaptive mechanism of bat, the suppression subtractive hybridization (SSH) library was constructed from the brain tissue of greater horseshoe bats. Dot blot screening was carried out and the up-regulated genes in hibernating state were obtained. Then RT-PCR and RQ-PCR were performed to test the expression patterns of selected cDNAs. Here we first show that the functional and non-functional isoforms of bat CaMKKbeta1 display distinct expression patterns between hibernating and active states. The up-regulation of non-functional form of CaMKKbeta1 may represent a new neuroprotective strategy adopted by bats or even other hibernators to avoid the CNS damage during hibernation. Our results showed that bat CaMKKbeta1 gene has four transcript isoforms and these transcript variants differ primarily in exons b and d, which are 129 bp and 43 bp respectively. Statistical analyses indicated that these isoforms display distinct expression patterns at different states, in which only isoform 3, the non-functional form, increased 300% at hibernating state. These results suggest that distinct expression patterns of transcript isoforms of a gene, which have different activity, may represent a new potential adaptive mechanism in hibernation, except for the simple up-regulation of selected genes/proteins and the reversible protein phosphorylation.