Circadian expression of Glycoprotein 2 (Gp2) gene is controlled by a molecular clock in mouse Peyer's patches

Abstract

The expression levels of many cell‐surface proteins vary with the time of day. Glycoprotein 2 (Gp2), specifically expressed on the apical surface of M cells in Peyer's patches, functions as a transcytotic receptor for mucosal antigens. We report that cAMP response element‐binding protein (CREB) regulates the transcription of the Gp2 gene, thereby generating the circadian change in its expression in mouse Peyer's patches. The transcytotic receptor activity of Gp2 was increased during the dark phase when the Gp2 protein abundance increased. Rhythmic expression of clock gene mRNA was observed in mouse Peyer's patches, and expression levels of Gp2 mRNA also exhibited circadian oscillation, with peak levels during the early dark phase. The promoter region of the mouse Gp2 gene contains several cAMP response elements (CREs). Chromatin immunoprecipitation assays revealed that CREB bound to the CREs in the Gp2 gene in Peyer's patches. Forskolin, which promotes CREB phosphorylation, increased the transcription of the Gp2 gene in Peyer's patches. As phosphorylation of CREB protein was increased when Gp2 gene transcription was activated, CREB may regulate the rhythmic expression of Gp2 mRNA in Peyer's patches. These findings suggest that intestinal immunity is controlled by the circadian clock system.

Oral Administration of Mice with Cell Extracts of Recombinant Lactococcus lactis IL1403 Expressing Mouse Receptor Activator of NF-kB Ligand (RANKL)

“…Isolation of RNA and synthesis of cDNA was same as above method. qRT-PCR was conducted using TB Green ® Premix Ex Taq TM (Tli RNaseH Plus, Takara Bio) with M cell-specific primers GP2-F:5′- GATACTGCACAGACCCCTCCA-3′; GP2-R:5′- GCAGTTCCGGTCATTGAGGTA-3′ ( Kusunose et al., 2020 ), and primers for the housekeeping gene GAPDH-F:5′-AACTTTGGCATTGTGGAAGGGCTC-3′; GAPDH-R:5′-AAGGCCATGCCAGTG AGCTTC-3′. mRNA levels were normalized to those of GAPDH ( Kim et al, 2015 ).…”