The use of lentivirus-mediated gene delivery to investigate the role of the Brain specific transcription factor MYT1L In-vitro and In-vivo

The Myt1l gene, which encodes a neuron-specific transcription factor, is highly<br/>expressed in the developing brain, suggesting a crucial role in the processes of<br/>neurodevelopment. Although the expression of this gene decreases after birth, it<br/>continues to have a good level of expression throughout adulthood in localized<br/>areas. Nevertheless, the role of this gene in neural development and resultant<br/>behaviours has not been firmly established yet. In an effort to elucidate its function, lentiviral vectors containing synthetic microRNA-adapted short hairpin (shRNAmir) targeting Myt1l were produced to significantly decrease its expression. The knockdown efficiency of these vectors was corroborated in-vitro in the human neural stem cell line (SPC04) and the mouse cell line Neuro-2A. The effects of downregulating MYT1L at gene expression level during differentiation of SPC04 were assessed at two time points: pre-differentiation and day 7 differentiation. The results from this experiment identified five genes being co-expressed with MYT1L during stem cell differentiation: BCL11B, MYT1, SYN1, SNAP25 and JPH3. Further in-silico analysis localized a MYT1L binding site in the promoter region of each one of those genes, suggesting that they might be direct targets of MYT1L. In order to expand our understanding of the possible behavioural effects of this gene, an invivo study was also performed in which adult mice received microinjections of lentiviruses expressing Myt1l shRNAmir in the dorsal hippocampus. These mice manifested a very subtle and transient increase in anxiety-like behaviour, which normalised with time. Even though the dorsal hippocampus has been only weakly linked to anxiety, our results seem to support this association when the expression of Myt1l is reduced. The findings of this thesis have laid the foundations of understanding the function of MYT1L pathway and the impact it has on behaviour

Name dc:type.qualificationname

Doctor of Philosophy

Level dc:type.qualificationlevel

Doctoral Thesis

Grantor dc:publisher.institution

King's College London

Year dc:date.issued

2014

Author dc:creator

• Martinez Medina, Lourdes

Advisor dc:contributor.advisor

• Desrivieres, Sylvane

Language dc:language

eng

Identifier

oai:kclpure.kcl.ac.uk:studenttheses/f293fc7c-fdac-48e4-a5d4-d5ca1d50ec03

OAI identifier oai:identifier

oai:kclpure.kcl.ac.uk:studenttheses/f293fc7c-fdac-48e4-a5d4-d5ca1d50ec03