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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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Western blot analysis of ald, hsp60, Tcp-1η and idh.y-axis represents relative expression level normalized by β-actin, x-axis represents different development stages on day 13, 15, 17, 19 and 20, accordingly. Different letters (a, b, c) are significantly different (p<0.05).
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pone-0020428-g010: Western blot analysis of ald, hsp60, Tcp-1η and idh.y-axis represents relative expression level normalized by β-actin, x-axis represents different development stages on day 13, 15, 17, 19 and 20, accordingly. Different letters (a, b, c) are significantly different (p<0.05).

Mentions: Among the 4 major groups (carbohydrate and energy production, protein folding, development and cytoskeleton) involved as key nodes in the BIN, 9 proteins were selected for further validating the proteins differential expressions at mRNA levels (Fig. 9) of which 4 of them were further confirmed by Western blot analysis (Fig. 10). The selected proteins were, ald (spot d17), pglym78 (spot u12), idh (spot u9), hsp60 (spot d9), hsp83 (spot d6), hsc70-4 (spot u2), l(1)g0022 (spot d12), Tcp-1η (spot d13) and tm2 (spot u21). Accordingly, 6 proteins showed consistent mRNA expressions with the change patterns of their corresponding proteins as in 2-DE gels. The 6 proteins that showed consistent increased mRNA abundance were ald (spot d17), hsp60 (spot d9), hsp90 (spot d6), l(1)g0022 (spot d12), tm2 (spot u21) and Tcp-1η (spot d13) and they were gene transcripts from early to middle stage (Fig. 9). However, the transcripts of genes of pglym (spot u12), idh (spot u9) and hsc70-4 (spot u2) showed variations between the mRNA transcription and protein abundance across the 5 time points and this might be due to lack of a direct relationship between mRNA timing and protein expressions and/or other regulatory mechanisms such as lack of synchronization. The result of the Western blot analysis also showed considerable expressional difference for ald (spot d17), hsp60 (spot d9), Tcp-1η (spot d13) and idh (spot u9) and the achieved differences were in line as in the 2-DE image (Fig. 10).


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)

Western blot analysis of ald, hsp60, Tcp-1η and idh.y-axis represents relative expression level normalized by β-actin, x-axis represents different development stages on day 13, 15, 17, 19 and 20, accordingly. Different letters (a, b, c) are significantly different (p<0.05).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0020428-g010: Western blot analysis of ald, hsp60, Tcp-1η and idh.y-axis represents relative expression level normalized by β-actin, x-axis represents different development stages on day 13, 15, 17, 19 and 20, accordingly. Different letters (a, b, c) are significantly different (p<0.05).
Mentions: Among the 4 major groups (carbohydrate and energy production, protein folding, development and cytoskeleton) involved as key nodes in the BIN, 9 proteins were selected for further validating the proteins differential expressions at mRNA levels (Fig. 9) of which 4 of them were further confirmed by Western blot analysis (Fig. 10). The selected proteins were, ald (spot d17), pglym78 (spot u12), idh (spot u9), hsp60 (spot d9), hsp83 (spot d6), hsc70-4 (spot u2), l(1)g0022 (spot d12), Tcp-1η (spot d13) and tm2 (spot u21). Accordingly, 6 proteins showed consistent mRNA expressions with the change patterns of their corresponding proteins as in 2-DE gels. The 6 proteins that showed consistent increased mRNA abundance were ald (spot d17), hsp60 (spot d9), hsp90 (spot d6), l(1)g0022 (spot d12), tm2 (spot u21) and Tcp-1η (spot d13) and they were gene transcripts from early to middle stage (Fig. 9). However, the transcripts of genes of pglym (spot u12), idh (spot u9) and hsc70-4 (spot u2) showed variations between the mRNA transcription and protein abundance across the 5 time points and this might be due to lack of a direct relationship between mRNA timing and protein expressions and/or other regulatory mechanisms such as lack of synchronization. The result of the Western blot analysis also showed considerable expressional difference for ald (spot d17), hsp60 (spot d9), Tcp-1η (spot d13) and idh (spot u9) and the achieved differences were in line as in the 2-DE image (Fig. 10).

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