Article Figures & Data
Figures
- Fig. 1.
Luminal catecholamine (CA) concentrations in the cecum of specific pathogen-free (SPF) mice. A representative of chromatogram data of luminal contents of the cecum is shown. Peaks NE, E and DA indicate norepinephrine, epinephrine, and dopamine, respectively.
- Fig. 2.
Luminal and tissue CA concentration in the gastrointestinal tract of SPF mice. Luminal (n = 11–15; A) and tissue (n = 5–8; B) CA levels were measured by the method described in methods. Open and closed circles show NE (left vertical axis) and DA (right vertical axis) levels, respectively. ***Significantly higher than the corresponding ileum value (P < 0.001).
- Fig. 3.
Free, glucuronide, and sulfate-conjugated CA in the gut lumen of SPF mice. Luminal glucuronide- and sulfate-conjugated DA (n = 6; A) or NE (n = 6; B) levels in the ileum, cecum, and colon were analyzed by the methods described in methods. The mean value of each form of CA is expressed as a percentage of the total (free CA + conjugated CA). The open, closed, and dotted bars show free, glucuronide-, and sulfate-conjugated CA, respectively.
- Fig. 4.
Cecal free CA levels in the lumen and tissue of SPF and germ-free (GF) mice. Luminal (n = 5; A) and tissue (n = 6–8; B) free CA levels in the SPF mice and GF mice were measured as described in methods. Luminal NE levels (left vertical axis) in the SPF and GF mice were 35 ± 5*** ng/g and 3.8 ± 1.3 ng/g, respectively; luminal DA levels (right vertical axis) in the SPF and GF mice were 115 ± 14*** ng/g and 5.0 ± 0.5 ng/g, respectively. ***Significantly higher than the corresponding GF value (P < 0.001). Tissue NE levels (left vertical axis) in the SPF and GF mice were 310 ± 24 ng/g and 290 ± 19 ng/g, respectively; tissue DA levels (right vertical axis) in the SPF and GF mice were 25 ± 4 ng/g and 22 ± 3 ng/g, respectively.
- Fig. 5.
Free glucuronide- and sulfate-conjugated CA in the gut lumen of GF mice. Luminal glucuronide- and sulfate-conjugated DA (A) or NE (B) levels in the ileum, cecum, and colon were analyzed according to the methods described in methods. The mean value of each form of CA is expressed as a percentage of the total (free CA + conjugated CA). The open, closed, and dotted bars show free, glucuronide-conjugated, and sulfate-conjugated CA, respectively.
- Fig. 6.
Luminal free CA levels and β-glucuronidase (GUS) activity in the cecum of gnotobiotic mice. A: cecal luminal contents from EX-GF (n = 6) and Clostridia (n = 6)-associated mice were processed for free NE and DA measurement, as described in methods. Free NE concentrations in the GF, EX-GF, and Clostridia-mice were 2.6 ± 0.3 ng/g, 18 ± 1 ng/g***, and 27 ± 3 ng/g***; free DA concentrations were 16.3 ± 0.7 ng/g, 120 ± 16 ng/g***, and 114 ± 10 ng/g***. ***P < 0.001 was significantly higher than the corresponding GF value. B: cecal luminal contents of GF, EX-GF, and Clostridia-associated mice were subjected to the measurement of GUS activity, as described in methods. GUS activity in the GF, EX-GF, and Clostridia-mice were 6 ± 0.3 μg PheP·h−1·mg protein−1, 110 ± 8 μg PheP·h−1·mg protein−1***, and 173 ± 16 μg PheP·h−1·mg protein−1***. ***P < 0.001 was significantly higher than the corresponding GF value.
- Fig. 7.
Free, glucuronide- and sulfate-conjugated CA in the cecal lumen of the mice colonized with E. coli mutant strain JW1609 or its parent strain BW25113. The cecal contents were subjected to the measurement of free, glucuronide-conjugated, and sulfate-conjugated NE (A) and DA (B) levels 4 wk after exposure to E. coli mutant strain devoid of GUS (n = 5, JW1609: ΔGUS) or its parent strain (n = 5, BW25113). The GF and SPF data shown in this figure are identical with ones shown in Tables 1 and 2. The mean value of each form of CA is expressed as a percentage of the total (free CA + conjugated CA). The open, closed, and dotted bars show free, glucuronide-conjugated, and sulfate-conjugated CA, respectively.
Tables
- Table 1.
Free, glucuronide-conjugated, and sulfate-conjugated CA in gut lumen of SPF mice
Free CA, ng/g Glucuronide-Conjugated CA, ng/g Sulfate-Conjugated CA, ng/g Ileum NE 9.0 ± 2.1 3.8 ± 1.9 1.7 ± 1.2* DA 15.7 ± 4.1 44.9 ± 5.1‡ 4.7 ± 1.4 Cecum NE 34.6 ± 4.6 0.5 ± 0.2† 1.5 ± 0.4† DA 115.4 ± 14.4 1.6 ± 1.0† 4.1 ± 1.5† Colon NE 60.5 ± 6.0 2.0 ± 0.5† 0.6 ± 0.2† DA 177.0 ± 10.9 2.5 ± 0.7† 0.3 ± 0.2† Values are means ± SE. Luminal glucuronide- and sulfate-conjugated norepinephrine (NE) and dopamine (DA) levels in the gut lumen of specific pathogen-free (SPF) mice (n = 6) were measured according to the methods described in methods. CA, catecholamines. Significant difference from the corresponding free NE or DA value:
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↵* P < 0.05;
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↵† P < 0.001;
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↵‡ P < 0.001 were significantly different from the corresponding free NE or DA value.
- Table 2.
Free, glucuronide-conjugated, and sulfate-conjugated CA in gut lumen of GF mice
Free CA, ng/g Glucuronide-Conjugated CA, ng/g Sulfate-Conjugated CA, ng/g Ileum NE 7.5 ± 3.7 1.9 ± 0.3 0.4 ± 0.1 DA 1.5 ± 0.3 130.1 ± 16.3† 3.6 ± 0.7 Cecum NE 3.8 ± 1.3 2.8 ± 0.6 0.4 ± 0.1* DA 5.0 ± 0.5 136.9 ± 29.7† 6.6 ± 1.6 Colon NE 3.2 ± 0.6 2.8 ± 0.6 0.4 ± 0.1* DA 4.8 ± 0.3 138.0 ± 20.1† 7.0 ± 1.1 - Table 3.
Free, glucuronide-conjugated, and sulfate-conjugated CA in cecal lumen of BW25113- and JW1609-inoculated mice
Free CA, ng/g Glucuronide-Conjugated CA, ng/g Sulfate-Conjugated CA, ng/g BW25113 NE 2.8 ± 0.4 0.6 ± 0.2 0.3 ± 0.1 DA 72.3 ± 8.8 31.5 ± 2.5 7.3 ± 0.8 JW1609 (ΔGUS) NE 2.2 ± 0.3 1.1 ± 0.3 0.3 ± 0.1 DA 31.9 ± 4.3* 87.9 ± 3.4† 6.9 ± 1.0
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