10 A niversary of IIMCB
10 A niversary of IIMCB
10 A niversary of IIMCB
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Fig. 2. 21 days in vitro hippocampal culture from rat embryos E19; stimulated with NMDA; Duolink in situ Proximity Ligation Assay for Calmyrin 2 and<br />
NSF interaction (red dots); nucleus (blue) (author: Magdalena Błażejczyk).<br />
in primary cultures <strong>of</strong> rat neurons is regulated by NMDA<br />
receptor activation and associated Ca 2+ signaling (Blazejczyk<br />
et al., submitted). We have also identified several new<br />
potential targets <strong>of</strong> CaMy2 in rat brains and confirmed<br />
these interactions by several methods in vitro. Physiological<br />
significance <strong>of</strong> these interactions in primary neurons is<br />
currently under investigation.<br />
4. Role and regulation <strong>of</strong> β-catenin in mature neurons<br />
(Katarzyna Misztal, Wojciech Michowski, Andrzej Nagalski,<br />
Marta Wiśniewska in collaboration with partners from<br />
PROMEMORIA 6th FP <strong>of</strong> EU)<br />
β-catenin is involved in the regulation <strong>of</strong> proliferation<br />
and differentiation <strong>of</strong> neuronal precursor cells as an<br />
activator <strong>of</strong> the Lef1/Tcf transcription factor and component<br />
<strong>of</strong> the cadherin cell-adhesion complex. In mature<br />
neurons β-catenin participates in synaptogenesis and<br />
synaptic function in the cadherin complex. However, its<br />
transcriptional activity in mature neurons remain elusive. We<br />
are interested in the function <strong>of</strong> β-catenin in the adult brain,<br />
since new data suggests it might be involved in learning and<br />
memory formation, as well as in some brain pathology. We<br />
look for β-catenin/Lef1 target genes in mature neurons i)<br />
and explore the mechanism <strong>of</strong> stabilization <strong>of</strong> β-catenin in<br />
mature thalamic neurons ii).<br />
i)Using Real Time PCR, immunohistochemistry and<br />
Western blot techniques we demonstrated that β-catenin<br />
and Lef1 are expressed at high levels in neurons <strong>of</strong><br />
the adult thalamus, in contrast to other regions <strong>of</strong> the<br />
forebrain. Moreover, both proteins are present in the cell<br />
nuclei, implying their involvement in gene expression. To<br />
answer the question about the role <strong>of</strong> the Lef1/β-catenin<br />
complex in the adult thalamic neurons we investigated a<br />
possible involvement <strong>of</strong> the complex in regulating genes<br />
encoding proteins indispensable for thalamic functions. We<br />
hypothesized that the Lef1/β-catenin transcription complex<br />
enhances expression <strong>of</strong> Cacna1G encoding Cav3.1 in mature<br />
Laboratory <strong>of</strong> Neurodegeneration 43