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Learn MoreWe utilized translating ribosome affinity purification (TRAP) coupled with RNA sequencing to examine mRNAs of GnRH neurons in adult intact and gonadectomized (GDX) male and female mice.; TRAP produces one RNA fraction enhanced for GnRH neuron transcripts and one RNA fraction depleted. cDNA libraries were created from each fraction and 50-base, paired-end sequencing done and differential expression; (enhanced fraction/depleted fraction) determined with a threshold of >1.5 or <0.66 fold (false discovery rate p0.05). A core of ~840 genes were differentially expressed in GnRH neurons in all treatments, including enrichment for Gnrh1 (~40 fold),; and genes critical for GnRH neuron and/or gonadotrope development. In contrast, non-neuronal transcripts were not enriched or were de-enriched. Several epithelial markers were also enriched,; consistent with the olfactory epithelial origins of GnRH neurons. Interestingly, many synaptic transmission pathways were de-enriched, in accordance with relatively low innervation of GnRH neurons.; The most striking difference between intact and GDX mice of both sexes was a marked down regulation of genes associated with oxidative phosphorylation and upregulation of glucose transporters in GnRH neurons from GDX mice. SOURCE: Richard,C,McEachin (mceachin@umich.edu) - University of Michigan
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