Long-Term Memory Testing
Objective: Assessment of long-term memory enhancement in HDAC3-FLOX mice and RGFP136-treated mice compared to wild-type controls
Gather these items before starting the experiment. Check off items as you prepare.
Equipment1
Not specified • Not specified • Not specified • Not specified
Materials3
Software1
Not specified • Not specified
As an Amazon Associate, we earn from qualifying purchases. Product links help support this free resource.
Protocol Steps
Generate focal HDAC3 deletions
Use HDAC3-FLOX genetically modified mice in combination with adeno-associated virus-expressing Cre recombinase to generate focal homozygous deletions of Hdac3 in area CA1 of the dorsal hippocampus
Note: This is a genetic approach to HDAC3 manipulation
View evidence from paper
“We used HDAC3-FLOX genetically modified mice in combination with adeno-associated virus-expressing Cre recombinase to generate focal homozygous deletions of Hdac3 in area CA1 of the dorsal hippocampus”
Administer RGFP136 pharmacologically
Use selective inhibitor of HDAC3, RGFP136, via intrahippocampal delivery as a complementary pharmacologic approach to HDAC3 manipulation
Note: This is a pharmacologic approach complementing the genetic approach
View evidence from paper
“To complement this approach, we also used a selective inhibitor of HDAC3, RGFP136”
Assess histone acetylation
Perform immunohistochemistry to determine if focal deletion or intrahippocampal delivery of RGFP136 resulted in increased histone acetylation
Note: Confirms molecular effects of HDAC3 manipulation
View evidence from paper
“Immunohistochemistry showed that focal deletion or intrahippocampal delivery of RGFP136 resulted in increased histone acetylation”
Test long-term memory
Assess long-term memory in HDAC3-FLOX mice and RGFP136-treated mice compared to wild-type controls to determine if memory is enhanced
Note: Both focal deletion of HDAC3 and HDAC3 inhibition via RGFP136 significantly enhanced long-term memory in a persistent manner
View evidence from paper
“Both the focal deletion of HDAC3 as well as HDAC3 inhibition via RGFP136 significantly enhanced long-term memory in a persistent manner”
Analyze gene expression
Examine expression of genes implicated in long-term memory from dorsal hippocampal punches using quantitative reverse transcription-PCR
Note: Expression of Nr4a2 and c-fos was significantly increased in HDAC3-FLOX mice compared with wild-type controls
View evidence from paper
“Next we examined expression of genes implicated in long-term memory from dorsal hippocampal punches using quantitative reverse transcription-PCR”
Test Nr4a2 mechanism
Deliver Nr4a2 small interfering RNA intrahippocampally to determine if Nr4a2 is required for memory enhancement observed in HDAC3-FLOX mice
Note: Memory enhancements observed in HDAC3-FLOX mice were abolished by intrahippocampal delivery of Nr4a2 siRNA
View evidence from paper
“Memory enhancements observed in HDAC3-FLOX mice were abolished by intrahippocampal delivery of Nr4a2 small interfering RNA”