retrograde raav pgk cre  (Addgene inc)

Journal: eLife

Article Title: Short-term social isolation acts on hypothalamic neurons to promote social behavior in a sex- and context-dependent manner

doi: 10.7554/eLife.94924

Figure Lengend Snippet: ( A ) Experimental timeline and viral strategy to chemogenetically inhibit the activity of POA social neurons in single-housed females. ( B ) Total time spent in resident-initiated social investigation is shown on saline and CNO days for 4 experimental groups: (red symbols) experimental females in which hM4Di is expressed in POA social neurons; (black symbols) control females in which GFP is expressed in POA social neurons; (brown symbols) control females in which hM4Di is expressed in ‘TRAPed’ AH neurons; (gray symbols) control females in which hM4Di is expressed in ‘TRAPed’ VMH neurons. ( C ) Same as ( B ), for proportion of trials with resident-initiated mounting. ( D ) Same as ( B ), for total USVs. ( E ) Total movement is plotted for females with hM4Di expressed in POA social neurons, on saline days vs. CNO days.

Article Snippet: The following viruses and injection volumes were used: AAV2/1-hSyn-FLEX-hM4Di-mCherry (Addgene #44262, 200 nL), AAV2/1-CAG-FLEX-EGFP-WPRE (Addgene #51502, 200 nL), AAV2/5-Ef1alpha-FLEX-taCasp3-TEVp (Addgene #45580, 200 nL), AAV2/1-Ef1alpha-hChR2(h134R)-EYFP-WPRE-HGHpA (Addgene #20298, 200 nL), AAVrg-pgk-Cre (Addgene #24593, 200 nL), and AAV2/5-hSyn-hM4Di-mCherry (Addgene #50475, 200 nL).

Techniques: Activity Assay, Saline, Control

Journal: eLife

Article Title: Short-term social isolation acts on hypothalamic neurons to promote social behavior in a sex- and context-dependent manner

doi: 10.7554/eLife.94924

Figure Lengend Snippet: ( A ) Time spent by POA social -hM4Di females engaged in social investigation during the TRAPing session is compared to the subsequent change in social investigation in the test sessions (CNO minus saline). ( B ) Same as ( A ), for time spent mounting. ( C ) Same as ( A ), for total USVs. ( D ) Experimental timeline and viral strategy to chemogenetically inhibit the activity of POA neurons TRAPed in single-housed females that were not given a social interaction. ( E ) Total time spent in resident-initiated social investigation is shown on saline and CNO days. ( F ) Same as ( E ), for proportion of trials with resident-initiated mounting. ( G ) Same as ( E ), for total USVs.

Article Snippet: The following viruses and injection volumes were used: AAV2/1-hSyn-FLEX-hM4Di-mCherry (Addgene #44262, 200 nL), AAV2/1-CAG-FLEX-EGFP-WPRE (Addgene #51502, 200 nL), AAV2/5-Ef1alpha-FLEX-taCasp3-TEVp (Addgene #45580, 200 nL), AAV2/1-Ef1alpha-hChR2(h134R)-EYFP-WPRE-HGHpA (Addgene #20298, 200 nL), AAVrg-pgk-Cre (Addgene #24593, 200 nL), and AAV2/5-hSyn-hM4Di-mCherry (Addgene #50475, 200 nL).

Techniques: Saline, Activity Assay

Journal: eLife

Article Title: Short-term social isolation acts on hypothalamic neurons to promote social behavior in a sex- and context-dependent manner

doi: 10.7554/eLife.94924

Figure Lengend Snippet: ( A ) Total time spent in resident-initiated social investigation is shown for group-housed male residents (teal) and single-housed male residents (maroon) during interactions with either female visitors (left) or male visitors (right). ( B ) Same as ( A ), for proportion of trials with resident-initiated mounting. ( C ) Same as ( A ), for total USVs recorded from pairs containing group-housed or single-housed male residents. ( D ) Total number of Fos-positive POA neurons is shown for group-housed male residents (teal) and single-housed male residents (maroon) following interactions with female visitors (left) or male visitors (right). ( E ) Experimental timeline and viral strategy to chemogenetically inhibit the activity of POA social neurons in single-housed males. ( F ) Total time spent in resident-initiated social investigation is shown on saline and CNO days for experimental males in which hM4Di is expressed in POA social neurons (red symbols, n=10) and control males in which GFP is expressed in POA social neurons (black symbols, n=10). ( G ) Same as ( F ), for proportion of trials with resident-initiated mounting. ( H ) Same as ( F ), for total USVs. All error bars show standard deviation.

Article Snippet: The following viruses and injection volumes were used: AAV2/1-hSyn-FLEX-hM4Di-mCherry (Addgene #44262, 200 nL), AAV2/1-CAG-FLEX-EGFP-WPRE (Addgene #51502, 200 nL), AAV2/5-Ef1alpha-FLEX-taCasp3-TEVp (Addgene #45580, 200 nL), AAV2/1-Ef1alpha-hChR2(h134R)-EYFP-WPRE-HGHpA (Addgene #20298, 200 nL), AAVrg-pgk-Cre (Addgene #24593, 200 nL), and AAV2/5-hSyn-hM4Di-mCherry (Addgene #50475, 200 nL).

Techniques: Activity Assay, Saline, Control, Standard Deviation

Journal: eLife

Article Title: Short-term social isolation acts on hypothalamic neurons to promote social behavior in a sex- and context-dependent manner

doi: 10.7554/eLife.94924

Figure Lengend Snippet: ( A ) Total time spent in resident-initiated social investigation during 30-min TRAPing session interactions with novel females is shown for male (red, n=9) and female (black, n=16) POA social -hM4Di mice. ( B ) Same as ( A ), for time spent in resident-initiated mounting. ( C ) Same as ( A ), for total USVs. Please note that n=2 POA social -hM4Di males and n=1 POA social -hM4Di female were excluded because their TRAPing session videos were not saved due to experimenter error. All error bars show standard deviation.

Article Snippet: The following viruses and injection volumes were used: AAV2/1-hSyn-FLEX-hM4Di-mCherry (Addgene #44262, 200 nL), AAV2/1-CAG-FLEX-EGFP-WPRE (Addgene #51502, 200 nL), AAV2/5-Ef1alpha-FLEX-taCasp3-TEVp (Addgene #45580, 200 nL), AAV2/1-Ef1alpha-hChR2(h134R)-EYFP-WPRE-HGHpA (Addgene #20298, 200 nL), AAVrg-pgk-Cre (Addgene #24593, 200 nL), and AAV2/5-hSyn-hM4Di-mCherry (Addgene #50475, 200 nL).

Techniques: Standard Deviation

Journal: eLife

Article Title: Short-term social isolation acts on hypothalamic neurons to promote social behavior in a sex- and context-dependent manner

doi: 10.7554/eLife.94924

Figure Lengend Snippet:

Article Snippet: The following viruses and injection volumes were used: AAV2/1-hSyn-FLEX-hM4Di-mCherry (Addgene #44262, 200 nL), AAV2/1-CAG-FLEX-EGFP-WPRE (Addgene #51502, 200 nL), AAV2/5-Ef1alpha-FLEX-taCasp3-TEVp (Addgene #45580, 200 nL), AAV2/1-Ef1alpha-hChR2(h134R)-EYFP-WPRE-HGHpA (Addgene #20298, 200 nL), AAVrg-pgk-Cre (Addgene #24593, 200 nL), and AAV2/5-hSyn-hM4Di-mCherry (Addgene #50475, 200 nL).

Techniques: Transfection, Construct, Recombinant, Software