spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

First published online May 1, 2006
Journal of Experimental Biology 209, 1988-1995 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02193
This Article
Right arrow Summary Freely available
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Folwell, J. L.
Right arrow Articles by Shepherd, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Folwell, J. L.
Right arrow Articles by Shepherd, D.

Immunolocalisation of the D. melanogaster Nramp homologue Malvolio to gut and Malpighian tubules provides evidence that Malvolio and Nramp2 are orthologous

James L. Folwell1, C. Howard Barton2,* and David Shepherd2

1 Institute of Genetics, The University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, UK
2 School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton SO16 7PX, UK


Figure 1
View larger version (66K):

[in a new window]
  		Fig. 1. Reactivity of anti-Mvl antiserum and demonstration of the specificity of blocking peptides. Western blots of purified wild-type GST, Mvl expressed in Cos-1 cells by transient transfection, and mock transfectants (–ve), were probed with anti-Mvl antiserum incubated overnight without peptide (A) or incubated with 25 µg of each peptide (B). Proteins were detected by ECL and size markers of the standard proteins. (Below) The amido black stain of the two membranes following immune detection.

 

Figure 2
View larger version (84K):

[in a new window]
  		Fig. 2. Immunolocalisation of Mvl in the stage-15 embryo. Mvl-positive staining appears in a subset of cells scattered throughout the haemocoel between the developing gut (g) and the epidermis of the embryo (e). These scattered cells have large phagosomes (arrows).(A) The subcellular localisation of Mvl appears to be adjacent to the phagosome. (B) Mvl-positive staining in two discrete regions, both associated with the phagosome-like structures. (C) Mvl peptides compete out anti-Mvl staining. Scale bars, 100 µm.

 

Figure 3
View larger version (99K):

[in a new window]
  		Fig. 3. Immunolocalisation of Mvl in the larval brain (A–C) and testis (D–F). (A) Reactivity in a subset of neurons within the larval brain. (B) Higher magnification of these neurons (arrows) shows staining associated with a sub-cellular compartment. (C) Immuno-blocking with Mvl peptides. (D) Reactivity within the larval gonad, and also staining associated with cells of the fat body (arrow). (E) Higher magnification show Mvl-positive staining associated with a subcellular structure within the cells of the testis (arrow). (F) Immuno-blocking with Mvl peptides. Scale bars, 200 µm (A,C,D,F); 100 µm (B,E).

 

Figure 4
View larger version (166K):

[in a new window]
  		Fig. 4. Immunolocalisation of Mvl in both the larval and adult alimentary canal. (A–C) Larval anterior midgut. (A) Mvl staining within the larval anterior midgut. (B) Higher magnification reveals punctuate subcellular staining within enterocytes (arrow). (D–F) Mvl-positive staining within the adult posterior midgut. (D) Staining within these enterocytes appears to be diffuse (arrow), with some punctate staining. (E) Small scattered cells show strong Mvl staining (arrow). (G–I) Mvl localisation within the Malpighian tubules. (G) Diffuse staining within principal (type 1) cells (arrow). (H) Strong Mvl-positive staining within small neuroendocrine-like cells (arrow). (C,F,I) Mvl peptides confirm staining specificity in each cell type. Scale bars, 200 µm (A,C); 100 µm (B,D–I).

 

Figure 5
View larger version (130K):

[in a new window]
  		Fig. 5. Immunolocalisation of Mvl in the adult brain and gonad. (A) Mvl-positive staining in neurons of the adult brain. (C) Mvl staining within a subset of cells in the accessory gland of the adult gonad (arrow). (B,D) Peptide block of staining. Scale bars, 20 µm.

 

Figure 6
View larger version (114K):

[in a new window]
  		Fig. 6. Reactivity of anti-Mvl antiserum with Drosophila protein extracts. (A) Western blot of protein extracts from embryo (lane 1), larvae (lane 2), pupae (lane 3), adult (lane 4), probed with Mvl antiserum. Two strong bands appear of approximately 42 kDa, and 50 kDa, respectively. (B) Amido black-stained membranes show equal loading.

 




© The Company of Biologists Ltd 2006