TRAPSeq identifies transcripts enriched/de-enriched in mouse GnRH neurons.

We 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 p≤0.05). 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. Exmaination of transcripts enriched in GnRH neurons in female, male, and gonadectomized mice (of both sexes) by TRAPSeq.