Fig. 1
From: Translational changes induced by acute sleep deprivation uncovered by TRAP-Seq
TRAP approach efficiently enriches for ribosome-associated mRNA transcripts from excitatory neurons in the mouse hippocampus. a CaMKIIα-Cre mice were crossed with Cre responsive Rpl22-HA mice. Male and female progeny (CaMKIIα-Cre:RiboTag) were used for all subsequent experiments. b Immunofluorescence images using anti-HA antibody (green) and DAPI (blue) from CaMKIIα-Cre:RiboTag mice shows expression of Rpl-22-HA expression in mouse hippocampus from male and female mice aged 2–3 months. All images have the same magnification with the scale bar shown indicating 500 microns. c CaMKIIα-Cre:RiboTag mice were either sleep deprived for 5 h (from ZT0 to ZT5) or non-sleep deprived, and hippocampus were dissected and processed for ribosome affinity purification of transcripts using anti-HA antibody. d, e qPCR analysis of immuno-pulldown efficiency revealed enrichment of CaMKIIα (excitatory neuronal marker gene) compared to GAD1 (interneuron marker gene) and GFAP (glial marker gene) in CamKIIα-Cre:RiboTag males (d) and females (e). Males: Mixed-effect analysis revealed no significant interaction between “Condition (SD or NSD)” and “Genes (IP enrichment)” F (2, 39) = 0.6336, p = 0.5361. Significant main effect of “Genes” F (1.024, 19.97) = 131.9, p < 0.0001 was observed, while no significant main effect of “condition” F (1, 22) = 0.6307, p = 0. 0.4356 was observed. Sidak multiple comparisons revealed significant difference between: GFAP vs. CaMKIIα: p < 0.0001, GAD1 vs. CaMKIIα, p < 0.0001. Females: Mixed-effect analysis revealed significant interaction between “Condition (SD or NSD)” and “Genes” F (2, 26) = 4.338, p = 0.0237. Significant main effect of “Genes”: F (1.055, 13.72) = 149.9, p < 0.0001, while no significant main effect of “Condition”: F (2, 39) = 0.6336, p = 0.5361 was observed. Sidak multiple comparisons revealed significant difference between: GFAP vs. CaMKIIα, p < 0.0001, GAD1 vs. CaMKIIα, p < 0.0001