Anxiety-Related Behavior Assessment
Objective: To measure anxiety-related behavioral responses in rats following vasopressin V1 receptor knockdown via antisense oligodeoxynucleotide administration into the septum
Gather these items before starting the experiment. Check off items as you prepare.
Equipment1
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Materials5
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Software1
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Protocol Steps
Surgical implantation of osmotic minipump
Implant osmotic minipump containing antisense oligodeoxynucleotide into male rats for infusion into the septum
Note: Chronic administration for up to 4 days
View evidence from paper
“infused an antisense oligodeoxynucleotide to the V1 subtype mRNA into the septum of male rats with osmotic minipumps”
Behavioral testing for social discrimination
Test the ability of animals to distinguish a previously exposed juvenile from a novel one
Note: Compared between antisense-treated, vehicle control, and scrambled-sequence oligo control groups
View evidence from paper
“diminished the ability of the animals to distinguish a previously exposed juvenile from a novel one”
Behavioral testing for anxiety-related responses
Measure anxiety-related behavior in treated animals
Note: Anxiety-related behavior was reduced in antisense-treated animals compared to controls
View evidence from paper
“anxiety-related behavior was reduced”
Exogenous AVP administration test
Administer exogenous vasopressin intracerebroventricularly and measure response in social memory
Note: Antisense-treated animals showed reduced ability to respond to exogenous AVP with improved social memory
View evidence from paper
“respond to exogenous AVP (1 ng/5 microliters, intracerebroventricular) with an improved social memory”
Receptor autoradiography
Measure radiolabeled AVP binding in septum and other limbic brain areas of behaviorally tested rats
Note: Antisense treatment resulted in reduced binding in septum but not other limbic areas
View evidence from paper
“As measured in the behaviorally tested rats, antisense treatment resulted in a reduced binding of radiolabeled AVP in the septum, but not in other limbic brain areas (receptor autoradiography)”
Reverse transcriptase PCR analysis
Measure V1 receptor mRNA levels in brain tissue from treated animals
Note: Antisense treatment resulted in increased V1 receptor mRNA, indicating translational arrest and ongoing transcriptional activity
View evidence from paper
“increased amount of V1 receptor mRNA (reverse transcriptase PCR), indicating translational arrest and ongoing transcriptional activity”