Table 1.
Characteristics of the primary and secondary antibodies used in immunoreactions.
Table 2.
Average area of the small intestine and correction factors applied for the of neuronal density correction in the myenteric and submucosal plexuses of the jejunum and ileum.
Experimental groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG). n = 8 rats per group.
Table 3.
Physiological parameters assessed in experimental groups.
Final weight body (FW), food intake (FI), tumor mass (TM), weight variation (WV). Experimental groups: control (C), control supplemented with L-glutamine (CG), Walker 256 tumor (TW) and Walker 256 tumor supplemented with L-glutamine (TWG). n = 8 rats per group.
Table 4.
Neuronal density of myenteric neurons (neurons/cm2) in the jejunum and ileum.
Experimental groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG). HuC/D-IR population (neurons/cm2), CHAT-immunoreactive subpopulation (neurons/cm2).
Table 5.
Neuronal density of submucous neurons (neurons/cm2) in the jejunum and ileum.
Experimental groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG). HuC/D-IR population (neurons/cm2) and VIP-immunoreactive subpopulation (neurons/cm2) densities.
Fig 1.
Representative images of Hu/CHAT myenteric neurons.
Double immunostaining of HuC/D and CHAT myenteric neurons of ileum (a-d) and jejunum (e-h). Experimental groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG). These images are composites of merged images taken separately from red (CHAT) and green (HuC/D) fluorescent channels. Scale Bar 50 μm.
Fig 2.
Representative photomicrographs of Hu/VIP submucous neurons.
Double immunostaining of HuC/D and VIP submucous neurons of jejunum (a-d) and ileum (e-f) from experimental groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG). These images are composites of merged images taken separately from red (VIP) and green (HuC/D) fluorescent channels. Scale Bar 50 μm.
Table 6.
Neuronal morphometric analysis of myenteric plexus.
Neuronal cell body area (μm2) of Jejunum and ileum from groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG).
Table 7.
Neuronal morphometric analysis of submucosal plexus.
Neuronal cell body area (μm2) of jejunum and ileum from groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG).
Fig 3.
Representative images of the myenteric varicosities nerve fibers.
Myenteric VIP-IR and CGRP-IR varicosities of the jejunum: (a-d) VIP-IR nerve fiber and (i-l) CGRP-IR nerve fiber. Ileum: (e-h) VIP-IR nerve fiber and (m-p) CGRP-IR nerve fiber. Experimental groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG). (q-x) representative images of the submucous CGRP-IR varicosities nerve fibers of the jejunum (q-t) and ileum (u-x) from experimental groups (C, CG, TW and TWG). All enlarge images on the top right of each image with white arrows indicate examples of immunoreactive varicosity (a'-x'). Scale Bar 25 μm.
Table 8.
Morphometric analysis of VIP-immunoreactive and CGRP-immunoreactive varicosities areas (μm2) in the myenteric plexus in the intestinal regions (jejunum and Ileum).
Experimental groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG).
Table 9.
Morphometric analysis of CGRP-immunoreactive varicosities areas (μm2) in the submucosal plexus in the jejunum and Ileum from the following groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG).
Fig 4.
Effect of supplementation of L-glutamine and cachexia on proteins (CHAT, VIP and CGRP) expression from jejunal and ileal samples of each experimental group.
Groups: control (C); control supplemented with 2% L-glutamine (CG); Walker-256 tumor (TW); and Walker-256 tumor supplemented with 2% L-glutamine (TWG). (a) Jejunum CHAT expression and representative bands (on the top). (b) Ileum CHAT expression and representative bands (on the top). (c) Jejunum VIP expression and representative bands (on the top). (d) Ileum VIP expression and representative bands (on the top). (e) Jejunum CGRP expression and representative bands (on the top). (f) Ileum CGRP expression and representative bands (on the top). Bars represent means ± SEM of samples from four animals. The data are presented as percentage arbitrary units (% of control) after normalization (GAPDH). * indicates significant difference (p < 0.05) versus control group.