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Distribution of elongation factor-1alpha in larval tissues of the fall armyworm, Spodoptera frugiperda.

Habibi J, Goodman CL, Stuart MK - J. Insect Sci. (2006)

Bottom Line: We determined the distribution of EF-1alpha in larval tissues of the fall armyworm, Spodoptera frugiperda , with a monoclonal antibody generated to EF-1alpha from Sf21 cells, a cell line developed from ovarian tissue of S. frugiperda.To a certain extent, EF-1alpha concentrations reflected the expected metabolic activity level of each of the represented tissues.Closer examination by immunofluorescence microscopy revealed that EF-1alpha concentrations varied among different cell types within a given tissue, i.e. midgut columnar epithelial cells yielded strong signals, while goblet cells failed to react with the EF-1alpha-specific antibody.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, University of Missouri, Columbia, MO, USA. habibij@health.missouri.edu

ABSTRACT
Elongation factor-1alpha (EF-1alpha) promotes the delivery of aminoacyl-tRNA to the acceptor site of the ribosome during protein synthesis. The enzyme has a number of additional functions, including regulation of apoptosis and interaction with the cytoskeleton. We determined the distribution of EF-1alpha in larval tissues of the fall armyworm, Spodoptera frugiperda , with a monoclonal antibody generated to EF-1alpha from Sf21 cells, a cell line developed from ovarian tissue of S. frugiperda. Enzyme-linked immunosorbent assay showed that EF-1alpha comprised 1.9-9.9% of the total protein within the tissues that were examined, which included fat body, Malpighian tubules, midgut, muscle, salivary glands, trachea, and ventral nerve cord. To a certain extent, EF-1alpha concentrations reflected the expected metabolic activity level of each of the represented tissues. Closer examination by immunofluorescence microscopy revealed that EF-1alpha concentrations varied among different cell types within a given tissue, i.e. midgut columnar epithelial cells yielded strong signals, while goblet cells failed to react with the EF-1alpha-specific antibody.

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Immunofluorescence staining of paraffin-embedded thick sections of midgut tissue from the second instar Spodoptera frugiperda. Strong immunofluorescent signals were observed in panels A and D (anterior section), and B and E (posterior section) of the larval midgut, after incubation with Mab 7D6 (primary antibody), followed by goat anti-mouse IgG conjugated to Cy5 (secondary antibody). No immunofluorescent signals were observed in panels C and F, incubated with blocking agent and secondary antibody alone. Panels on the left are fluorescent images, while panels on the right are fluorescent images overlaid onto light microscopy images of the same sections. BM = Basement membrane, EP = epithelial cell, GC = goblet cell, H = hemolymph, L = lumen, M = Malpighian tubules. Scale bar = 20 μm.
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i1536-2442-6-33-1-f03: Immunofluorescence staining of paraffin-embedded thick sections of midgut tissue from the second instar Spodoptera frugiperda. Strong immunofluorescent signals were observed in panels A and D (anterior section), and B and E (posterior section) of the larval midgut, after incubation with Mab 7D6 (primary antibody), followed by goat anti-mouse IgG conjugated to Cy5 (secondary antibody). No immunofluorescent signals were observed in panels C and F, incubated with blocking agent and secondary antibody alone. Panels on the left are fluorescent images, while panels on the right are fluorescent images overlaid onto light microscopy images of the same sections. BM = Basement membrane, EP = epithelial cell, GC = goblet cell, H = hemolymph, L = lumen, M = Malpighian tubules. Scale bar = 20 μm.

Mentions: Immunofluorescence microscopy of S. frugiperda second instars revealed variations in the amount of EF-1α expressed by different cell types within a given tissue (Fig. 3). Notably, the apical surfaces of midgut columnar epithelial cells yielded strong immunofluorescent signals, especially the brush-border microvilli, while goblet cells and basement membrane demonstrated no reactivity with Mab 7D6. Of particular interest is the expression of EF-1α within the larval midgut, because accumulation of EF-1α correlates with induction of apoptosis (Ruest et al. 2002; Schwientek et al. 2002; Duttaroy et al. 1998), and because apoptosis of the midgut epithelium is known to be an effective defense against baculovirus infection in the larval stages (Clem 2005). These observations have led us to speculate that the natural abundance of EF-1α in midgut epithelial cells might prime the cells for apoptosis following viral infection. It has been postulated that high levels of EF-1α enhance translation of pro-apoptotic factors such as caspase enzymes (Duttaroy et al. 1998), or cause distortion of the cell cytoskeleton by severing microtubules (Kato 1999), thus leading to cell death.


Distribution of elongation factor-1alpha in larval tissues of the fall armyworm, Spodoptera frugiperda.

Habibi J, Goodman CL, Stuart MK - J. Insect Sci. (2006)

Immunofluorescence staining of paraffin-embedded thick sections of midgut tissue from the second instar Spodoptera frugiperda. Strong immunofluorescent signals were observed in panels A and D (anterior section), and B and E (posterior section) of the larval midgut, after incubation with Mab 7D6 (primary antibody), followed by goat anti-mouse IgG conjugated to Cy5 (secondary antibody). No immunofluorescent signals were observed in panels C and F, incubated with blocking agent and secondary antibody alone. Panels on the left are fluorescent images, while panels on the right are fluorescent images overlaid onto light microscopy images of the same sections. BM = Basement membrane, EP = epithelial cell, GC = goblet cell, H = hemolymph, L = lumen, M = Malpighian tubules. Scale bar = 20 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2990326&req=5

i1536-2442-6-33-1-f03: Immunofluorescence staining of paraffin-embedded thick sections of midgut tissue from the second instar Spodoptera frugiperda. Strong immunofluorescent signals were observed in panels A and D (anterior section), and B and E (posterior section) of the larval midgut, after incubation with Mab 7D6 (primary antibody), followed by goat anti-mouse IgG conjugated to Cy5 (secondary antibody). No immunofluorescent signals were observed in panels C and F, incubated with blocking agent and secondary antibody alone. Panels on the left are fluorescent images, while panels on the right are fluorescent images overlaid onto light microscopy images of the same sections. BM = Basement membrane, EP = epithelial cell, GC = goblet cell, H = hemolymph, L = lumen, M = Malpighian tubules. Scale bar = 20 μm.
Mentions: Immunofluorescence microscopy of S. frugiperda second instars revealed variations in the amount of EF-1α expressed by different cell types within a given tissue (Fig. 3). Notably, the apical surfaces of midgut columnar epithelial cells yielded strong immunofluorescent signals, especially the brush-border microvilli, while goblet cells and basement membrane demonstrated no reactivity with Mab 7D6. Of particular interest is the expression of EF-1α within the larval midgut, because accumulation of EF-1α correlates with induction of apoptosis (Ruest et al. 2002; Schwientek et al. 2002; Duttaroy et al. 1998), and because apoptosis of the midgut epithelium is known to be an effective defense against baculovirus infection in the larval stages (Clem 2005). These observations have led us to speculate that the natural abundance of EF-1α in midgut epithelial cells might prime the cells for apoptosis following viral infection. It has been postulated that high levels of EF-1α enhance translation of pro-apoptotic factors such as caspase enzymes (Duttaroy et al. 1998), or cause distortion of the cell cytoskeleton by severing microtubules (Kato 1999), thus leading to cell death.

Bottom Line: We determined the distribution of EF-1alpha in larval tissues of the fall armyworm, Spodoptera frugiperda , with a monoclonal antibody generated to EF-1alpha from Sf21 cells, a cell line developed from ovarian tissue of S. frugiperda.To a certain extent, EF-1alpha concentrations reflected the expected metabolic activity level of each of the represented tissues.Closer examination by immunofluorescence microscopy revealed that EF-1alpha concentrations varied among different cell types within a given tissue, i.e. midgut columnar epithelial cells yielded strong signals, while goblet cells failed to react with the EF-1alpha-specific antibody.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, University of Missouri, Columbia, MO, USA. habibij@health.missouri.edu

ABSTRACT
Elongation factor-1alpha (EF-1alpha) promotes the delivery of aminoacyl-tRNA to the acceptor site of the ribosome during protein synthesis. The enzyme has a number of additional functions, including regulation of apoptosis and interaction with the cytoskeleton. We determined the distribution of EF-1alpha in larval tissues of the fall armyworm, Spodoptera frugiperda , with a monoclonal antibody generated to EF-1alpha from Sf21 cells, a cell line developed from ovarian tissue of S. frugiperda. Enzyme-linked immunosorbent assay showed that EF-1alpha comprised 1.9-9.9% of the total protein within the tissues that were examined, which included fat body, Malpighian tubules, midgut, muscle, salivary glands, trachea, and ventral nerve cord. To a certain extent, EF-1alpha concentrations reflected the expected metabolic activity level of each of the represented tissues. Closer examination by immunofluorescence microscopy revealed that EF-1alpha concentrations varied among different cell types within a given tissue, i.e. midgut columnar epithelial cells yielded strong signals, while goblet cells failed to react with the EF-1alpha-specific antibody.

Show MeSH
Related in: MedlinePlus