Quantitative Assessment of Macrophages in the Muscularis Externa of Mouse Intestines

TY - JOUR

T1 - Quantitative Assessment of Macrophages in the Muscularis Externa of Mouse Intestines

AU - Mikkelsen, H B

AU - Larsen, J O

AU - Froh, P

AU - Nguyen, T H

N1 - Copyright © 2011 Wiley-Liss, Inc.

PY - 2011/9

Y1 - 2011/9

N2 - Quantification of intestinal cells is challenging for several reasons: The cell densities vary throughout the intestines and may be age dependent. Some cell types are ramified and/or can change shape and size. Additionally, immunolabeling is needed for the correct identification of cell type. Immunolabeling is dependent on both up- and down-regulation of the antigen being labeled as well as on the primary and secondary antibodies, the fixation, and the enhancement procedures. Here, we provide a detailed description of immunolabeling of CD169+ cells and major histocompatibility class II antigen (MHCII+) cells and the subsequent quantification of these cells using design-based stereology in the intestinal muscularis externa. We used young (5-weeks-old) and adult (10-weeks-old) mice. Cell densities were higher in jejunum-ileum, when compared with colon. In jejunum/ileum, the cell densities increased in oral-anal direction in adults, whereas the densities were highest in the midpart in young animals. In colon, the cell densities decreased in oral-anal direction in both groups of animals. Except for the density of MHCII+ cells in colon, the cell densities were highest in young animals. Densities of CD169+ and MHCII+ cells did not differ, except in the colon of young animals where the CD169+ density was almost twice as high as the MHCII+ density. CD169 and MHCII antigens seem to be expressed simultaneously by the same cell in jejunum/ileum. We conclude that cell densities depend on both the age of the mouse and on the location in the intestines.

AB - Quantification of intestinal cells is challenging for several reasons: The cell densities vary throughout the intestines and may be age dependent. Some cell types are ramified and/or can change shape and size. Additionally, immunolabeling is needed for the correct identification of cell type. Immunolabeling is dependent on both up- and down-regulation of the antigen being labeled as well as on the primary and secondary antibodies, the fixation, and the enhancement procedures. Here, we provide a detailed description of immunolabeling of CD169+ cells and major histocompatibility class II antigen (MHCII+) cells and the subsequent quantification of these cells using design-based stereology in the intestinal muscularis externa. We used young (5-weeks-old) and adult (10-weeks-old) mice. Cell densities were higher in jejunum-ileum, when compared with colon. In jejunum/ileum, the cell densities increased in oral-anal direction in adults, whereas the densities were highest in the midpart in young animals. In colon, the cell densities decreased in oral-anal direction in both groups of animals. Except for the density of MHCII+ cells in colon, the cell densities were highest in young animals. Densities of CD169+ and MHCII+ cells did not differ, except in the colon of young animals where the CD169+ density was almost twice as high as the MHCII+ density. CD169 and MHCII antigens seem to be expressed simultaneously by the same cell in jejunum/ileum. We conclude that cell densities depend on both the age of the mouse and on the location in the intestines.

U2 - 10.1002/ar.21444

DO - 10.1002/ar.21444

M3 - Journal article

C2 - 21809459

SN - 1932-8486

VL - 294

SP - 1557

EP - 1565

JO - Anatomical Record

JF - Anatomical Record

IS - 9

ER -