These results confirmed our microarray data. Transcripts unique to the TRH neurons To further breakdown the microarray data, a second method of analysis of the original signal intensities derived from the MAS software analysis was performed using a stringent P value. This approach allowed us to identify transcripts present in the different cell popula tions with a high degree of certainty. Using a P value of 0. 001, we identified a total of 1864 and 1701 tran scripts whose Inhibitors,Modulators,Libraries presence in the two GFP replicates was significant. Similarly, we identified 1776 and 1714 tran scripts in the replicate samples for the GFP cell popu lation. In the NT cell population, we identified 1925 transcripts.

In order to identify the transcripts that were present in both replicates and to reduce the false positive rate in the detection Inhibitors,Modulators,Libraries of expressed transcripts, we defined the repre sentative set of each sample as that containing transcripts significantly expressed in both replicates according to the P 0. 001 threshold. This resulted in 1600 transcripts as representative of the GFP cell population and 1630 transcripts for the GFP cell population. As shown in Figure 3, the over lap between the three cell populations indicates that 1361 transcripts were common to the three populations, whereas 112 transcripts were common to the GFP and GFP cell populations but not expressed in the NT cell population. This comparison also shows that 51 tran scripts were unique to the GFP cell population, while 50 transcripts were unique to the GFP cell population at these thresholds.

It should be noted Carfilzomib that in this con text unique transcripts refer to those transcripts that are uniquely detected in one or more populations shown in the Venn diagram, as they are likely to be expressed in undetectable levels at these thresholds in other compared populations. According to their GenBank Inhibitors,Modulators,Libraries annotations, several of these 50 transcripts are related to neuronal phenotype, e. g. synaptojanin 1, to translation machinery, e. g. eukaryotic transla tion initiation factor 3 subunit 9, ribosomal pro tein L27, to basal metabolic machinery, e. g. acyl CoA synthetase long chain family member 5, solute carrier family 37 member 4, to cell sig naling, e. g. the serine threonine kinase 38, in addition, transcripts encoding proteins with RNA proces sing properties were also observed, i. e.

the nuclear tran scription factor Y gamma, Inhibitors,Modulators,Libraries the splicing factor arginine serine rich 10 and the Y box protein 1. Transcripts related to CNS development were also identified, i. e. neurofilament heavy chain polypeptide, and the nuclear factor I B. A transcript with chromatin remodeling properties, the transforma tion transcription domain associated protein was also identified. These transcripts may play a critical role in the fetal development of hypothalamic TRH neurons. Discussion Events occurring during development are tightly coupled to gene expression regulation.