Cristin-resultat-ID: 1140103
Sist endret: 23. juni 2014, 14:35
Resultat
Mastergradsoppgave
2014

Characterization of two predicted long non-coding RNAs in long-term synaptic plasticity in rat hippocampus

Bidragsytere:
  • Ida Sporild

Utgiver/serie

Utgiver

Universitetet i Bergen
NVI-nivå 0

Om resultatet

Mastergradsoppgave
Publiseringsår: 2014

Beskrivelse Beskrivelse

Tittel

Characterization of two predicted long non-coding RNAs in long-term synaptic plasticity in rat hippocampus

Sammendrag

Design: The project was initiated with deep RNA sequencing to reveal differential expressed RNA two hours after induction of LTP by high-frequency stimulation (HFS) of the medial perforant path input to the dentate gyrus. In this thesis, the expression and localization of two predicted lncRNAs were further investigated. The expression of lncRNAs in the dentate gyrus at different time-points post high-frequency stimulation (HFS) was determined by quantitative real-time PCR (qPCR). In situ hybridization was done using antisense RNA probes, to elucidate the cellular distribution and intracellular localization of lncRNA in rat brain sections. Fluorescent in situ hybridization (FISH) was used for detection of intracellular location of lncRNA1 in dentate gyrus two hours after HFS. The effect of BDNF treatment of primary hippocampal neuronal culture on the expression of lncRNA in in vitro was investigated with qPCR. To identify protein partners that can bind the lncRNA candidates, a biotin/streptavidin affinity pull-down method was used. Results: The dynamic expression of lncRNA1 and lncRNA2 in HFS-LTP was confirmed with Quantitative real-time PCR (qPCR). The induction of lncRNA1 and lncRNA2 was dependent on N-methyl-D-aspartate receptor (NMDAR) activation and specific to the dentate gyrus. The temporal expression profile at three different time points after LTP induction show that lncRNA1 expression peaks at two hours and lncRNA2 expression is still increasing after five hours. In addition, the lncRNA Gomafu that has previously been studied in neuronal cell cultures is decreased in LTP. Furthermore, the changes in lncRNA expression are restricted to the granule cell layer of the dentate gyrus. The expression of lncRNA1 and lncRNA2 tends to increase after BDNF treatment of primary hippocampal neuronal cultures. These findings suggest that lncRNA1 and lncRNA2 expression is regulated by synaptic activity both in vivo and in vitro. The predicted full-length transcripts for lncRNA1 and lncRNA2 were obtained by molecular cloning and will be used for future RNA-protein pull-down experiments. The method of RNA-protein pull-down was successfully carried out with a control RNA. Conclusion: Taken together, the work presented in this thesis contributes to identifying two novel lncRNA in the rat hippocampus. The two novel lncRNAs are confirmed to be dynamically expressed and temporally and spatially regulated in the dentate gyrus after LTP induction in vivo. This is the first work describing the temporal and spatial activity-dependent regulation of lncRNA in the context of activity-dependent in vivo. This research implicates the need of further characterization of lncRNA expression and functions, to disentangle the multifaceted mechanisms they exploit in gene regulation. These two RNAs might have important and distinct regulatory roles in activity-dependent synaptic plasticity. Thrilling questions about the connection between the expression and function of lncRNA remain to be solved.

Bidragsytere

Ida Sporild

  • Tilknyttet:
    Forfatter
    ved Institutt for biomedisin ved Universitetet i Bergen

Clive Raymond Evjen Bramham

Bidragsyterens navn vises på dette resultatet som Clive R. Bramham
  • Tilknyttet:
    Veileder
    ved Institutt for biomedisin ved Universitetet i Bergen

Karin Wibrand

  • Tilknyttet:
    Veileder
    ved Institutt for biomedisin ved Universitetet i Bergen
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