Alterations of stress related proteins in genetically altered mice revealed by two-dimensional differential in-gel electrophoresis analysis.
Transgenic, knockout and knockin mice are useful tools for linking specific genes with behaviour and other complex biological processes. However, complications arising due to compensatory changes, genetic background differences and other factors could lead to difficulty in interpreting the resulting changes in phenotype. We have used fluorescence two-dimensional differential in-gel electrophoresis in combination with matrix-assisted laser desorption/ionization-time of flight mass fingerprinting to investigate the possibility that distinct genetic alterations can lead to common protein expression changes in genetically modified mice. Brain proteomes were compared from two transgenic mouse strains (Tg2576 x TgPS1 and Tg2576), two knockout mouse strains (5-HT(7)R -/- and GABA(A)Ralpha5 -/-) and one knockin mouse strain (GABA(A)Ralpha1-H101R). Both of the transgenic models showed an isoform change in the heat shock 70 related protein, mortalin. The knockout and knockin models showed similar changes in mortalin expression along with an alteration of the anti-oxidant protein 2. The observed proteomic alterations indicate that stress-responsive protein pathways may be altered artefactually in all of the mouse models used in this study and highlights an area where caution is needed in interpreting proteomic changes in genetically modified mice.
Skynner
Heather A
HA
Neuroscience Research Centre, Merck, Sharp & Dohme Research Laboratories, Harlow, Essex, CM20 2QR, UK. heather_skynner@merck.com
Rosahl
Thomas W
TW
Knowles
Michael R
MR
Salim
Kamran
K
Reid
Lee
L
Cothliff
Rosa
R
McAllister
George
G
Guest
Paul C
PC
D000818:Animals; D001923:Brain Chemistry; D002232:Carbocyanines / Q000737:chemistry; D002352:Carrier Proteins; D016208:Databases, Factual; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005786:Gene Expression Regulation; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis; D006360:Heat-Shock Proteins / Q000032:analysis*; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D024101:Mitochondrial Proteins; D009363:Neoplasm Proteins / Q000032:analysis; D011963:Receptors, GABA-A / Q000032:analysis; D011985:Receptors, Serotonin / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D002232:Carbocyanines; D002352:Carrier Proteins; D018840:HSP70 Heat-Shock Proteins; C492001:HSPA9 protein, human; D006360:Heat-Shock Proteins; C083794:Hspa9 protein, mouse; D024101:Mitochondrial Proteins; D009363:Neoplasm Proteins; D011963:Receptors, GABA-A; D011985:Receptors, Serotonin; C080352:mortalin; C104425:serotonin 7 receptor