The Regulatory Factor ZFHX3 Modifies Circadian Function in SCN via an AT Motif-Driven Axis.
Bottom Line: Using RNA sequencing, we found minimal effects on core clock genes in Zfhx3(Sci/+) SCN, whereas the expression of neuropeptides critical for SCN intercellular signaling was significantly disturbed.Moreover, mutant ZFHX3 had a decreased ability to activate AT motifs in the promoters of these neuropeptide genes.Lentiviral transduction of SCN slices showed that the ZFHX3-mediated activation of AT motifs is circadian, with decreased amplitude and robustness of these oscillations in Zfhx3(Sci/+) SCN slices.
Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.Show MeSH
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Mentions: We used RNA sequencing to identify transcriptional targets of ZFHX3. RNA was extracted from SCN tissue punches from Zfhx3Sci/+ and Zfhx3+/+ animals at zeitgeber time (ZT)3 and ZT15 (n = 3 for each time by genotype combination). RNA sequencing revealed that 242 genes were differentially expressed at either one or both time points (log2 fold change > 1, p < 0.05) (Table S1), with 28 of those surviving multiple testing correction (q < 0.05 in at least two quantification methods). At this more stringent level, 19 genes were differentially expressed at ZT3 and 13 at ZT15, while 4 genes were affected at both time points (Figure 2A). The majority of genes (17 of 28) showed a decrease in expression in Zfhx3Sci/+ (Figure 2B). Interestingly, the expression of a number of circadian-related neuropeptides was decreased in Zfhx3Sci/+ SCN, including Vip, its receptor (Vipr2), and prokineticin receptor 2 (Prokr2) (Figures 2C–2E). We conducted gene ontology (GO) enrichment analysis using all of the differentially expressed genes (q < 0.05 in at least one quantification method, n = 169) and found significant enrichment for a number of GO terms including neuron differentiation, regulation of cellular metabolic process, and regulation of cell proliferation (Table S2).
Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.