Neuron-Derived Estrogen is Critical for Astrocyte Activation and Neuroprotection of the Ischemic Brain

We examined the transcriptional changes modulated in ischemic brain of fore brain specific aromatase knockout mice (FBN-ARO-KO) by performing global transcriptome analysis.Total RNA was isolated from FLOX and FBN-ARO-KO mice that underwent 20-minute CCA occlusion, followed by 24-hour reperfusion. The collected hippocampi samples were subjected to RNA isolation and Illumina TruSeq RNA sample preparation and sequencing was done using UT Health San Antonio genomics core facility protocol. We observed that some of the key pathways that regulate astrocyte reactivity, such as RhoA signaling, actin-based motility by Rho signaling, signaling by Rho family GTPases, and protein ubiquitination, NRF2-mediated oxidative stress response pathways were significantly altered in FBN-ARO-KO+GCI mice. Overall design: Total RNA was isolated from female FLOX and FBN-ARO-KO mice that underwent 20-minute CCA occlusion, followed by 24-hour reperfusion. RNA was isolated from the hippocampi using the RNeasy mini kit. Illumina TruSeq RNA sample preparation was performed following manufacturer's protocol. Samples were run on an Illumina HiSeq 3000 in duplicate. The combined raw reads were aligned to UCSC mm9 and genes were annotated by Tophat. Genes were annotated and quantified by HTSeq-DESeq pipeline.