Protein expression profiles in mice during spatial learning

Tan , Yee (2008) Protein expression profiles in mice during spatial learning. Masters thesis, Universiti Malaysia Sabah.

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Abstract

Learning and remembering what has been learnt (memory) is a very important process which enables organism to adapt to the challenges of the environment and survive. Due to active research in this field, many theories and hypotheses on the mechanisms of memory consolidation and the components involved in the process of learning and memory have been postulated. Proteins such as N-methyl-D-aspartic acid (NMDA), y-aminobutyrate (GABA), S-hydroxytryptamine and D1/D5 dopamine receptors, Calmodulin-dependent protein kinase II (CaMKII) and Ca²⁺ -dependent protein kinase C, have been discovered to be involved in the process. We have used two-dimensional gel electrophoresis to unravel some of the proteins involved in learning and memory. This research is a preliminary investigation into the proteomics underlying the process of spatial learning and memory in the brain. We have optimized the separation parameters to the extend of obtaining 1088 spots per gel, that were resolved sufficiently to be detected by the 20 analysis software, ImageMaster 20 platinum 6.0, GE Healthcare. Seventeen male mice aged from 8 to 12 weeks were used. Seven mice were subjected to spatial learning in a Morris water maze; 6 mice acted as quiet controls and 4 mice acted as swim controls. Total brain protein was extracted. The protein content was about 17.S mg/g brain. One hundred and seventy-six matches were identified with a minimum of 20% difference in protein expression between the trained and quiet control mice. However, these matches may be expressed not only due to the spatial learning process but also by the swimming exercise during the training in the Morris water maze. Among these 176 matches, there were 20 matches that had a maximum of 20% difference in protein expression between quiet and swim control mice. These 20 matches have a very high possibility of being proteins that are expressed preferentially during the spatial learning process. Of these 20 matches, there are 4 matches, 10 10004, 10789, 10313 and 11231 that had significant differences between trained mice and the swim controls as well as the quiet controls. Through search using database, UniProt Knowledgebase, based on estimated molecular weights, match ID 10004 was found to have molecular weight comparable with the 05 dopamine receptor, DrdS (fragment) (9.3 kDa). Match ID 10789 has a molecular weight comparable to Calcium/calmodulin-dependent kinase type I, CamK1, (41.6 kDa). Match 10 10313 has a molecular weight comparable with Fyn protein (60.0 kDa), calcium/calmodulin-dependent kinase kinase type II, CamKKII (60.6 kDa) and CalCium/calmodulin-dependent kinase type II, CamK2b (60.6 kDa). These proteins are known to be involved in the mechanism of memory formation. Match 10 11231 could not be associated with any matches of known proteins that are involved in the mechanism of memory formation. Further research Including the use of liquid chromatography with mass spectrophotometry or time of flight (LC-MSn or LC-TOF) and amino acid sequencing, should be continued to identify these proteins of interest, especially those spots in the 20 matches.

Item Type:Thesis (Masters)
Uncontrolled Keywords:gel electrophoresis, protein, spatial learning, memory
Subjects:Q Science > QP Physiology
Divisions:SCHOOL > School of Medicine
ID Code:10081
Deposited By:IR Admin
Deposited On:11 Dec 2014 14:21
Last Modified:11 Dec 2014 14:21

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