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First published online May 18, 2006
Journal of Experimental Biology 209, 2215-2223 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02249

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The regenerative nidi of the locust midgut as a model to study epithelial cell differentiation from stem cells

Irineu Illa-Bochaca and Luis M. Montuenga^*

Department of Histology and Pathology, Schools of Sciences and Medicine, University of Navarra, E-31080 Pamplona, Spain

View larger version (114K): [in a new window]   Fig. 1. Single BrdU administration experiment. Insects dissected 4 h (A) and 120 h (B) after the BrdU administration. Scale bar, 20 µm.

View larger version (16K): [in a new window]   Fig. 2. Representation of niche height and distance of labelled nuclei to the base of the epithelium distances plotted against time (N=20). Labelled distance adjusts significantly to a lineal regression (R=0.93) with the following formula y=0.39x+37.3.

View larger version (15K): [in a new window]   Fig. 3. Frequencies of the three different niche types (SN, MN and NN) over 8 days of experimentation in the single BrdU administration experiment (N=20). SN, niches with small positive nuclei; MN, niches with only medium-sized positive nuclei; NN, negative niches with no BrdU-stained cells.

View larger version (29K): [in a new window]   Fig. 4. Isometric vision of 3-D reconstructed image from confocal stack images. (A) BrdU-labelled nuclei (green) located inside the niche area. (B) FMRFamide-like cytoplasmatic detection of endocrine cells (red). (C) Superimposition of A and B; no colocalization was observed. Scale bar, 10 µm.

View larger version (16K): [in a new window]   Fig. 5. Number of BrdU-labelled cells in the niche after a single BrdU administration. Significant differences (P<0.05) were only observed when comparing Day 0 with the other three days (N=15). This suggests that endoreduplication may occur in the BrdU-labelled cells.

View larger version (116K): [in a new window]   Fig. 6. Continuous BrdU administration experiments. Insects corresponding to Group 1 (A) and Group 2 (B), which received a BrdU administration every 24 h or 48 h, respectively. Scale bar, 20 µm.

View larger version (16K): [in a new window]   Fig. 7. Percentage of labelled mitosis against time (N=144). The cell cycle phase duration (Tc) was estimated following the method described by Willems (Willems, 1972). Percentage of positive mitotic figures refers to the ratio between BrdU labelled and total (labelled + unlabelled) mitosis.

View larger version (129K): [in a new window]   Fig. 8. Insect dissected 14 days after the last BrdU administration. The arrowhead shows a long-term label-retaining cell located at the base of the niche. Scale bar, 10 µm.

View larger version (74K): [in a new window]   Fig. 9. Double immunocytochemistry, BrdU-labelled nuclei in brown and endocrine (FMRFamide-like) cells with pink cytoplasm. No colocalization was observed in the ventricle midgut area (A). However, BrdU-labelled nuclei can be clearly observed in some of the ampullar endocrine cells (B,C), some of which have double immunostaining for BrdU and FMRFa (C). Scale bar, 20 µm.