D & V Hipp. BrdU+ Cells by Age_Data Assoc. w. PMID37146791
Description
Electroconvulsive seizure, vs. sham procedure, was administered once daily for 10 consecutive days to paired male WT and Egr3-/- mice. Brain tissue was collected and cellular proliferation in the hippocampal formation was analyzed via bromodeoxyuridine (BrdU) immunohistochemistry. Analyses investigated whether number of BrdU+ cell counts were associated with animal age. Complete methods are found in associated Data in Brief article (submitted) and original associated article: Meyers, K.T., et al., Serial electroconvulsive Seizure alters dendritic complexity and promotes cellular proliferation in the mouse dentate gyrus; a role for Egr3. Brain Stimul, 2023. 16(3): p. 889-900. https://doi.org/10.1016/j.brs.2023.04.022 PMID: 37146791
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Mice were group housed in our AAALAC approved facility on a 14:10 hour light:dark cycle with ad libitum access to food and water. For BrdU studies, Egr3+/- mice that had been back-crossed to C57Bl/6 mice (from Taconic Laboratories) for > 15 generations were crossed to produce littermate experimental Egr3-/- and WT mice. Male mice were used for all studies. Electroconvulsive seizure (ECS) was administered to anesthetized mice (5% isoflurane) 1x/day for 10 consecutive days. Twenty-four hours following the tenth daily ECS, mice received the first of two injections of BrdU (100 mg/kg), spaced 24 hours apart (i.e., on days 11 and 12). Mice were returned to their home cage between procedures and sacrificed by isoflurane overdose 24 hours after the second BrdU injection (on day 13). Tissue was flash frozen in -40°C methylbutane. Brains from four conditions (WT no ECS, WT serial ECS, Egr3-/- no ECS, and Egr3-/- serial ECS) were placed in optimal cutting temperature (OCT) medium in a brain mold, frozen at -20°C, cut in 20 μm sections, and mounted on glass microscope slides together, to control for slide-to-slide variation. Antibody staining was performed with Anti-BrdU antibody with peroxidase fragments (Cat. 11585860001, Roche). Confocal microscope images were taken from six tissue sections per animal. The following regions of the hippocampal formation were analyzed: dentate gyrus superior (DGsp) and inferior (DGip) blades, including the subgranular zone (SGZ), of the left and right hemispheres from the dorsal (Bregma -1.34 mm to -2.35 mm) and ventral (Bregma -2.7 mm to -3.4 mm) poles. The number of BrdU positive cell profiles (representing recently divided cells) were counted and the average counts from all six sections per animal were calculated and represented per mm2 of tissue.
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Funding
National Institutes of Health
R01 MH097803
National Institutes of Health
R21 MH113154