Prenatal Androgenization Alters the Development of GnRH Neuron and Preoptic Area RNA Transcripts in Female Mice (II)

Translating Ribosome Affinity Purification results in non specific RNA binding to the immunoprecipitation matrix, even in mice not expressing cell specific GFP labelled ribosomal protein L10a. This creates false positive enriched transcripts in our datasets. To determine which RNA transcripts bind non specifically to the immunoprecipitation matrix, preoptic area punches were collected from young and adult mice that do not express L10a-GFP. Also, included was a positive control (GnRH CRE x floxed L10a-GFP mice) to ensure that the TRAP procedure was enriching for GnRH neuron specific transcripts as previously described. Four GnRH depleted supernatant RNA samples from the previous study were also included to aid comparison of the two sequencing runs. Overall design: There are 16 new RNA samples, 122429-122440. All except samples 122433 and 122438 are from L10a GFP negative mice and therefore negative control samples. 122432 and 122438 are from GnRH CRE-L10a GFP mice, therefore ribsomes in GnRH neurons are GFP labelled. This positive control biological replicate was included to ensure that the technique worked as it should. Samples 122441-122444 are technical replicates from the other data set (GSE155314). 122441 = 95718(GSM4698966), 122442 =95722(GSM4698970), 122443 = 95731(GSM4698979) and 122444 = 95734(GSM4698982).