Differential role of the vagus in diurnal gene expression rhythms in the small intestine

The objective of this study was to examine the function of vagal innervation in maintaining diurnal rhythmicity in the expression of intestinal absorptive genes. Rats underwent truncal vagotomy and were maintained for 7 days on nighttime scheduled feeding (12-h light/12-h dark cycle). Vagotomized rats (V; n = 9) were pair-fed with sham-operated controls (S; n = 4). Unoperated normal rats (N; n = 6) were also included as controls. Half the rats were killed 3 h after lights on (ZT3; Zeitgeber Time, with lights-on considered ZT0) and the other half at ZT9, the time interval over which we have previously shown that sucrase and sugar transporter expression exhibits a significant anticipatory increase. RNA and protein extracted from mucosa of proximal jejunums were subjected to Northern and Western blot analyses to assess the increase in gene expression. Sham operation did not alter the normal diurnal rhythmicity of intestinal gene expression. Control rats (S plus N) exhibited the expected increase in RNA levels at ZT9 versus ZT3 for SGLT1 (4.5-fold), GLUT2 (5.3-fold), GLUT5 (4.1-fold), and sucrase (2.9-fold; P > 0.001 in all cases). In contrast, the induction in V rats was markedly blunted for GLUT2 (1.3-fold) and sucrase (1.5-fold) but not for SGLT1 (5.0-fold) or GLUT5 (4.2-fold). The mRNA levels for GLUT2 and sucrase at ZT9 were significantly lower in V rats versus controls (P < 0.001). GLUT2 and SGLT1 protein levels exhibited a parallel pattern: SGLT1 induction was 4.3-fold in control rats (P < 0.01) and 3.8-fold in V rats (P <0.01), whereas GLUT2 induction was 3.3-fold in control rats (P < 0.01) but only 1.4-fold in V rats (NS). Our results indicate that signaling through the vagus nerve is necessary to maintain the anticipatory induction pattern of GLUT2 and sucrase. The persistent rhythm in both SGLT1 and GLUT5 indicates that (1) diurnal induction of these genes is independent of vagal innervation and (2) the procedure did not cause an overall loss of intestinal function. Thus, entrainment of anticipatory diurnal gene expression in the intestine occurs via two separate pathways that are differentially dependent on vagal input.