q-PCR

To measure doublecortin (DCX) mRNA levels in forebrain Ca_v1.2 cKO mice and AAV2-2/2-Cre-GFP injected cacna1c floxed (cacna1c ^fl/fl) mice, mice were euthanized by rapid decapitation and whole brains were rapidly dissected. Brain tissue was sectioned on a 1 mm brain block. Dentate gyrus-containing tissue punches were obtained from forebrain Cav1.2 cKO and wild-type mice. For AAV2/2-Cre-GFP and AAV2/2-GFP injected mice, GFP goggles (BLS) were used to visualize GFP signal in brain sections containing the dentate gyrus and to selectively dissect GFP-positive tissue. Tissue punches were processed for total RNA isolation using the mirVana RNA isolation kit (Life Technologies) and cDNA was synthesized from purified RNA using the High Capacity RNA-to-cDNA kit (Applied Biosystems). Cav1.2 mRNA levels were measured using cacna1c-specific primers (Qiagen QuantiTect Primer assay QT00150752), and DCX levels were measured using DCX-specific primers (Qiagen QuantiTect Primer assay QT02521155) on an ABI PRISM 7000 Sequence Detection System with SYBR Green PCR Master Mix (Applied Biosystems). Cycle threshold (Ct) values for target genes were normalized to the housekeeping gene gapdh (QuantiTect Primer assay QT01658692, Qiagen). Each experiment was performed in triplicate and values were averaged.