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Proteomic analysis of honeybee (Apis mellifera L.) pupae head development.

Zheng A, Li J, Begna D, Fang Y, Feng M, Song F - PLoS ONE (2011)

Bottom Line: Furthermore, the constructed protein interaction network predicted 33 proteins acting as key nodes of honeybee pupae head growth of which 9 and 4 proteins were validated at gene and protein levels, respectively.In this study, we uncovered potential protein species involved in the formation of honeybee pupae head development along with their specific temporal requirements.This first proteomic result allows deeper understanding of the proteome profile changes during honeybee pupae head development and provides important potential candidate proteins for future reverse genetic research on honeybee pupae head development to improve the performance of related organs.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture/Institute of Apicultural Research, Chinese Academy of Agricultural Science, Beijing, China.

ABSTRACT
The honeybee pupae development influences its future adult condition as well as honey and royal jelly productions. However, the molecular mechanism that regulates honeybee pupae head metamorphosis is still poorly understood. To further our understand of the associated molecular mechanism, we investigated the protein change of the honeybee pupae head at 5 time-points using 2-D electrophoresis, mass spectrometry, bioinformatics, quantitative real-time polymerase chain reaction and Western blot analysis. Accordingly, 58 protein spots altered their expression across the 5 time points (13-20 days), of which 36 proteins involved in the head organogenesis were upregulated during early stages (13-17 days). However, 22 proteins involved in regulating the pupae head neuron and gland development were upregulated at later developmental stages (19-20 days). Also, the functional enrichment analysis further suggests that proteins related to carbohydrate metabolism and energy production, development, cytoskeleton and protein folding were highly involved in the generation of organs and development of honeybee pupal head. Furthermore, the constructed protein interaction network predicted 33 proteins acting as key nodes of honeybee pupae head growth of which 9 and 4 proteins were validated at gene and protein levels, respectively. In this study, we uncovered potential protein species involved in the formation of honeybee pupae head development along with their specific temporal requirements. This first proteomic result allows deeper understanding of the proteome profile changes during honeybee pupae head development and provides important potential candidate proteins for future reverse genetic research on honeybee pupae head development to improve the performance of related organs.

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Global expression intensity map (hierarchical clustering) showing upregulation (red) and downregulation (green) across the ages of the pupae indicated on the top of each column with lists of functional proteins in the right column.
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pone-0020428-g006: Global expression intensity map (hierarchical clustering) showing upregulation (red) and downregulation (green) across the ages of the pupae indicated on the top of each column with lists of functional proteins in the right column.

Mentions: The hierarchical cluster analysis of differentially expressed proteins showed that 58 distinct proteins (excluding 1 protein with unknown functions) were partaken in the expression intensity map (Fig. 6). Generally, most of the protein spots behaved heterogeneously, but clustered under 2 large very homogenous expressional pictures that were from the early to middle stages (day 13–17) and the late developmental stage (day 19–20) with a shifting trend inline with the pupae head developmental stage. From 36 protein spots that were highly expressed during the early to middle stages, proteins as carbohydrate metabolism and energy production, development, protein biosythesis, cytoskeleton and protein folding were recognized as major groups (Fig. 6). To this fact, there were 7 proteins spots involved in carbohydrate metabolism and energy production (spots d14, d15, d17, d19, d20, d22 and d24), 6 in development (spots d10, d11, d12, d13, d30 and d31), 6 in biosythesis (spot d1–4, d5, d7), 5 in cytoskeleton (spots d25, d26, d27, d29 and d32) and 4 in protein folding (spots d6, d8, d9 and d28). In addition, there were 3 amino acid metabolism (spots d23, d36 and d33), 3 molecular transporters (spots d16, d34 and d35) and 2 nucleotide metabolism (spots d18, d21) that were upregulated during the early to middle stage. On the other hand, 22 proteins were upregulated during the late pupae stage that included 6 in development (spots u1, u3, u10, u11, u13 and u14), 5 in carbohydrate metabolism and energy production (spots u5, u6, u9, u12, and u17), 4 in cytoskeleton (spots u8, u15, u21 and u23) and 3 in proteins folding (spots u2, u4 and u22). The other proteins upregulated during late developmental stages included one involved in molecular transporter (spot u19); one related to protein biosynthesis (spot u7), one associated with fatty acid metabolism (spot u16) and one involved in antioxidant activity protein (spot u20) (Fig. 6).


Proteomic analysis of honeybee (Apis mellifera L.) pupae head development.

Zheng A, Li J, Begna D, Fang Y, Feng M, Song F - PLoS ONE (2011)

Global expression intensity map (hierarchical clustering) showing upregulation (red) and downregulation (green) across the ages of the pupae indicated on the top of each column with lists of functional proteins in the right column.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3102718&req=5

pone-0020428-g006: Global expression intensity map (hierarchical clustering) showing upregulation (red) and downregulation (green) across the ages of the pupae indicated on the top of each column with lists of functional proteins in the right column.
Mentions: The hierarchical cluster analysis of differentially expressed proteins showed that 58 distinct proteins (excluding 1 protein with unknown functions) were partaken in the expression intensity map (Fig. 6). Generally, most of the protein spots behaved heterogeneously, but clustered under 2 large very homogenous expressional pictures that were from the early to middle stages (day 13–17) and the late developmental stage (day 19–20) with a shifting trend inline with the pupae head developmental stage. From 36 protein spots that were highly expressed during the early to middle stages, proteins as carbohydrate metabolism and energy production, development, protein biosythesis, cytoskeleton and protein folding were recognized as major groups (Fig. 6). To this fact, there were 7 proteins spots involved in carbohydrate metabolism and energy production (spots d14, d15, d17, d19, d20, d22 and d24), 6 in development (spots d10, d11, d12, d13, d30 and d31), 6 in biosythesis (spot d1–4, d5, d7), 5 in cytoskeleton (spots d25, d26, d27, d29 and d32) and 4 in protein folding (spots d6, d8, d9 and d28). In addition, there were 3 amino acid metabolism (spots d23, d36 and d33), 3 molecular transporters (spots d16, d34 and d35) and 2 nucleotide metabolism (spots d18, d21) that were upregulated during the early to middle stage. On the other hand, 22 proteins were upregulated during the late pupae stage that included 6 in development (spots u1, u3, u10, u11, u13 and u14), 5 in carbohydrate metabolism and energy production (spots u5, u6, u9, u12, and u17), 4 in cytoskeleton (spots u8, u15, u21 and u23) and 3 in proteins folding (spots u2, u4 and u22). The other proteins upregulated during late developmental stages included one involved in molecular transporter (spot u19); one related to protein biosynthesis (spot u7), one associated with fatty acid metabolism (spot u16) and one involved in antioxidant activity protein (spot u20) (Fig. 6).

Bottom Line: Furthermore, the constructed protein interaction network predicted 33 proteins acting as key nodes of honeybee pupae head growth of which 9 and 4 proteins were validated at gene and protein levels, respectively.In this study, we uncovered potential protein species involved in the formation of honeybee pupae head development along with their specific temporal requirements.This first proteomic result allows deeper understanding of the proteome profile changes during honeybee pupae head development and provides important potential candidate proteins for future reverse genetic research on honeybee pupae head development to improve the performance of related organs.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture/Institute of Apicultural Research, Chinese Academy of Agricultural Science, Beijing, China.

ABSTRACT
The honeybee pupae development influences its future adult condition as well as honey and royal jelly productions. However, the molecular mechanism that regulates honeybee pupae head metamorphosis is still poorly understood. To further our understand of the associated molecular mechanism, we investigated the protein change of the honeybee pupae head at 5 time-points using 2-D electrophoresis, mass spectrometry, bioinformatics, quantitative real-time polymerase chain reaction and Western blot analysis. Accordingly, 58 protein spots altered their expression across the 5 time points (13-20 days), of which 36 proteins involved in the head organogenesis were upregulated during early stages (13-17 days). However, 22 proteins involved in regulating the pupae head neuron and gland development were upregulated at later developmental stages (19-20 days). Also, the functional enrichment analysis further suggests that proteins related to carbohydrate metabolism and energy production, development, cytoskeleton and protein folding were highly involved in the generation of organs and development of honeybee pupal head. Furthermore, the constructed protein interaction network predicted 33 proteins acting as key nodes of honeybee pupae head growth of which 9 and 4 proteins were validated at gene and protein levels, respectively. In this study, we uncovered potential protein species involved in the formation of honeybee pupae head development along with their specific temporal requirements. This first proteomic result allows deeper understanding of the proteome profile changes during honeybee pupae head development and provides important potential candidate proteins for future reverse genetic research on honeybee pupae head development to improve the performance of related organs.

Show MeSH
Related in: MedlinePlus