Circadian oscillation of innate immunity components in mouse small intestine

The digestive system is a major port of entry for pathogens. To detect and combat pathogens, the innate immunity in the gut utilizes pattern recognition receptors, such as Toll-like receptors (TLRs) and nucleotide-binding oligomerization domain (NOD) proteins, and broad-spectrum anti-bacterial polypeptides, such as defensins. We have previously shown that mouse enteric defensins (cryptdins) oscillate around the circadian cycle and peak at the end of the dark phase suggesting control by the biological clock. As the core mechanism of the biological clock has never been studied in the small intestine, our objective was to determine whether the biological clock is functional in mouse jejunum and examine whether mTlr and mNod2 mRNAs, similarly to cryptdins, oscillate throughout the circadian cycle. Mouse jejunum and Paneth-enriched crypt base cells were isolated around the circadian day and the levels of clock (mClock, mBmal1, mPer1, mPer2, mCry1) and innate immunity component (mTlr2, mTlr3, mTlr4, mTlr5, mTlr9, mNod2) genes were measured by real-time PCR. Analysis of mouse jejunum and Paneth-enriched crypt base cells revealed that all clock genes exhibited circadian oscillation. Similarly to cryptdins, mTlr2, mTlr3, mTlr4, mTlr5 displayed circadian rhythmicity in mouse jejunum. Although no circadian oscillation could be detected for mTlr9 and mNod2 in the whole jejunum, these genes oscillated in Paneth-enriched crypt base cells. In addition, mTlr3 exhibited the highest expression level. As the clock regulates intestinal motility and function, resetting of the clock in the small intestine may help not only to restore activity but also to gain better protection against pathogens.