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1

Lentiviral delivered siRNA against ICER protects primary cortical neurons from apoptosis caused by serum depriviation

100%
Klejman A., Bieganska K., Gierej D., Kaczmarek L.
Acta Neurobiologiae Experimentalis
|
2009
|
tom 69
|
nr 3
CREB activation and CREB-dependent signaling pathways are crucial for neuronal survival. The term ICER (inducible cAMP early repressor) refers to four protein isoforms that are all endogenous, inducible antagonists of CREB. It was previously shown, that all 4 ICER isoforms are induced upon pro-apoptotic treatment, and also that each of them separately evokes neuronal cell death in cortical culture transfected with these genes. The ICER proteins are believed to be strong repressors of Immediate Early Genes, which are involved in cell response to inter- and/or intra-cellular signals. Herein, we have applied the siRNA approach to silence ICER expression. Because ICERs are members of CREM family of proteins, sharing with them the gene sequence, only the small unique region for ICER was selected to design ICER-directed, specifi c siRNA. Indeed, we obtained functional siRNA capable of blocking ICERs but not affecting CREM proteins. With this tool, we have investigated if the ICER’s silencing protects neurons from apoptosis caused by either serum deprivation or excitotoxicity. Using the lentiviral vector, as a vehicle to deliver siRNA (shRNA) we have found that silencing of ICER mildly, although signifi cantly, protects primary cortical neurons from apoptosis caused by serum deprivation.
2

Sound frequency discrimination: functional role of c-FOS in auditory cortex

64%
Gierej D., Lioudyno V., Mikosz M., Nowak A., Blazejczyk M., Filipkowski R. K., Jaworski J., Kaczmarek L., Knapska E.
Acta Neurobiologiae Experimentalis
|
2011
|
tom 71
|
nr S
Immediate-early genes, c-fos in particular, are expressed in the brain upon neuronal activation. c-Fos expression is thought to reflect novelty detection and propensity for a plastic change as its levels decrease when the animal fully acquire a new task. However, its functional role in the brain remains largely unknown. In the present study, we used c-Fos immmunolabeling to identify the cortical network components within auditory and motor cortices that subserve sound frequency discrimination in a behavioral context. We designed a protocol in a fully automated cage for mice (IntelliCage), in which animals were supposed to discriminate between two sounds of different frequencies, one signaling safe access to water and another signaling punishment (air puff). c-Fos expression was analyzed at different stages of the training. Within the anterior parts of ventral and dorsal auditory cortices significant changes in c-Fos levels were observed that correlated with the discrimination learning. On the other hand, c-Fos expression in the motor cortex correlated only with motor activity. In order to address questions concerning the role of c-Fos in learning discrimination of sounds, associated with a particular behavioral meaning, RNA interference was used. c-Fos expression in the auditory cortex was blocked using short hairpin RNA (shRNA) delivered by lentiviral vectors. Mice were bilaterally injected with control vectors and harboring shRNA for c-fos (experimental). Then, the animals were subjected to sound discrimination training. Both the control and the experimental animals were responsive to the aversive cues, but the experimental group learnt much more slowly and showed faster extinction of the learned behavior than the control group. These results show the functional involvement of c-Fos within the auditory cortex in sound discrimination learning. More generally, they also support the idea that c-Fos marks neuronal plasticity rather than simple activation.
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