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The role of CRMP2 in the nervous system development.
Žiak, Jakub ; Balaštík, Martin (advisor) ; Jiruška, Přemysl (referee) ; Horák, Martin (referee)
Regulation of axon guidance and pruning of inappropriate synapses is key to development of neural circuits. Secreted semaphorins are integral part of both processes. Collapsin response mediator protein 2 (CRMP2) has been shown to regulate axon guidance by mediating Semaphorin 3A (Sema3A) signaling, however, nothing is known about its role in the synapse pruning. Similarly, it is also not known if CRMP2 mediates signals from other semaphorins. We herein studied CRMP2 protein and revealed its role in growth and pruning of selected axons and dendrites. In newly generated crmp2-/- and crmp2a-/- mice we demonstrate that CRMP2 has a moderate effect on Sema3A-dependent axon guidance in vivo, and its deficiency leads to a mild defect in axon guidance in peripheral nerves and corpus callosum. CRMP2A isoform is specifically involved in development of callosal axons. Surprisingly, we show that crmp2-/- mice display prominent defects in stereotyped axon pruning in hippocampus and visual cortex and altered dendritic spine remodeling, which are consistent with impaired Sema3F signaling and with models of autism spectrum disorder (ASD). Indeed, we demonstrate that CRMP2 mediates Sema3F signaling in primary neurons and that crmp2-/- mice display ASD-related social behavior changes in early postnatal period as well...
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Production of auxin transporter PIN1 in HEK293 cell line
Petermannová, Romana ; Vaněk, Ondřej (advisor) ; Hýsková, Veronika (referee)
PIN1 protein belongs to a PIN family of transporters. This group of transporters is significantly involved in auxin transports outside of the cell. The auxin transport among plant cells gives an important role in plant growth, gravitropism, etc. The principle of auxin transport by this carrier protein is still not explored. This work deals with the production of PIN1 carrier protein by recombinant expression in human embryonic kidney 293 cell line. Production in these cells was checked by the accumulation of radioactive auxin and by the immunofluorescence.
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