Abstract
Circulating short-chain fatty acids (SCFAs) are primarily derived from bacterial fermentation of carbohydrates in the colon where they function as physiologic modulators of epithelial cell maturation. Butyrate has been shown to induce tyrosine hydroxylase, the rate-limiting enzyme of catecholamine synthesis, and enkephalin neuropeptide gene transcription, suggesting a role in perinatal sympathoadrenal stress-adaptation. We sought to determine whether there were SCFA structural requirements for this effect. Nine biologically relevant SCFAs and butyrate derivatives were tested in an in vitro model (PC12, rat pheochromocytoma cells) for their ability to regulate neurotransmitter-related gene expression. Our results revealed that among all the studied SCFAs, only propionate and butyrate increased tyrosine hydroxylase and proenkephalin mRNA levels. The functional activity was selective to the carbon atom chain length and associated with the presence of an ethyl moiety in the carbon atom backbone chain. Modifications or absence of this domain affected the gene induction response, suggesting a receptor-mediated mechanism(s). Moreover, propionate, butyrate, and the drug 4-phenyl-butyrate were each shown to regulate transmitter genes via at least three independent mechanisms: histone hyperacetylation, cAMP signaling, or peroxisome proliferator-activated receptor gamma–mediated pathways. Thus, the biologic impact of SCFAs on catecholaminergic and opioid systems depend on the activation of SCFA-specific, dose-specific, and gene-specific molecular mechanisms. We speculate that 1) circulating levels of SCFAs may influence sympathoadrenal transmitter biosynthesis and hence whole animal stress-adaptive responsiveness after birth, and 2) the adverse effects of antibiotics on delayed acquisition of postnatal gut flora may affect this apparent evolutionary advantage of gut colonization.
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Abbreviations
- ppEnk:
-
preproenkephalin
- TH:
-
tyrosine hydroxylase (EC 1.14.16.2)
- PC12:
-
rat pheochromocytoma
- SCFA:
-
short-chain fatty acids
- SB:
-
sodium butyrate
- PKA:
-
protein kinase A
- PPAR:
-
peroxisome proliferator-activated receptor
- RT-PCR:
-
reverse transcriptase-PCR
- HDAC:
-
histone deacetylase
- TSA:
-
trichostatin A
- 4-PB:
-
phenylbutyrate
- 2-HB:
-
2-hydroxybutyrate
- 3-HB:
-
3-hydroxybutyrate
- RXR:
-
retinoid X receptor
- PPRE:
-
peroxisone proliferator response element
- PMSF:
-
phenylmethyl sulfenylfluoride
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Acknowledgements
The authors thank Ning Yuan (Regeneron Inc.) for the excellent technical assistance in some of the laboratory experiments.
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Supported by institutional grants from the Children's Foundation of the Department of Pediatrics.
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Mally, P., Mishra, R., Gandhi, S. et al. Stereospecific Regulation of Tyrosine Hydroxylase and Proenkephalin Genes by Short-Chain Fatty Acids in Rat PC12 Cells. Pediatr Res 55, 847–854 (2004). https://doi.org/10.1203/01.PDR.0000119365.21770.45
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DOI: https://doi.org/10.1203/01.PDR.0000119365.21770.45
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