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肠道碱性磷酸酶解毒脂多糖并预防炎症在肠道微生物群的响应
来源: | 作者:詹妮弗·M·贝茨1，Janie Akerlund，Erika Mittge，和Karen Guillemin | 发布时间: 2020-06-23 | 11290 次浏览 | 分享到:

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Figure 1.
Iap activity and iap transcription are regulated by LPS. In situ hybridization of iap transcript at 5 days post fertilization (dpf) in (A) a whole mount larva, and (B) a transverse section through the mid intestine (at the point indicated by the arrow in A). The iap specific purple stain is present in the intestinal epithelium (outlined with the dotted line in A) and is distinct from the black pigment cells above and below the digestive tract and the swim bladder (sb) in A. Scale bar in panel A = 100 μm, scale bar in panel B = 5 μm. (C) Semi-quantitative RT-PCR analysis showing iap and alp expression in dissected intestines (I) and carcasses from which intestines were removed (C) of 8 dpf larvae untreated or exposed to 50 μg/ml LPS for 24h. Levels of the housekeeping gene ef1α are shown as an amplification and loading control. (D) AP activity in the carcasses and intestines of 8 dpf untreated CV WT larvae, iap-MO injected larvae or larvae exposure to 10mM L-phen from 5 dpf. † Indicates values that differ significantly as compared to the CV levels of each group (carcass or intestine), P<0.01. (E) AP activity in 5 and 8 dpf intestines from larvae reared CV (solid bars) left untreated, exposed at 5 dpf to 30 μg/ml LPS, or injected at the 1 cell stage with myd88-MO or the control galT-MO, or larvae reared GF (striped bars) left untreated, exposed at 5 dpf to 3 μg/ml LPS, or mono-associated at 5 dpf with a Gram-negative Aeromonas species (G−) or a Gram-positive Streptococcus species (G+). † Indicates values that differ significantly from CV at 8dpf, * indicates values that differ significantly from GF at 8 dpf, P<0.01. For D and E, n=10 dissected intestines/treatment for each trial, with at least 2 trials per treatment. Error bars represent standard deviation. (F) iap transcript levels, measured by qRTPCR, were reduced in 8 dpf GF versus CV animals, and elevated in 8 dpf CV and GF animals exposed for 24h to 30 μg/ml LPS, but not in 8 dpf myd88-MO injected animals reared CV and exposed for 24h to 50 μg/ml LPS. Data are representative of two repeated trials, in which all samples were run in triplicate. Error bars indicate standard deviation. All animals were WT unless otherwise indicated.