Proteomic analysis of honeybee worker (Apis mellifera) hypopharyngeal gland development

Mao, Feng; Yu, Fang; Li, Jianke

doi:10.1186/1471-2164-10-645

Cited by 76 publications

(144 citation statements)

References 54 publications

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“…The centrality of these five protein families are also consistent with the high representation in the post-embryonic development of other honey bee organs, such as the pupal head (60), brain (61), hemolymph (62), hypopharyngeal gland (63,64), and salivary gland (65).…”

Section: Ligustica) Embryos (Normalized By Gapdh) (A) Is Significantsupporting

confidence: 69%

In-depth Proteomics Characterization of Embryogenesis of the Honey Bee Worker (Apis mellifera ligustica)

Mao

Han

et al. 2014

Molecular & Cellular Proteomics

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Identifying proteome changes of honey bee embryogenesis is of prime importance for unraveling the molecular mechanisms that they underlie. However, many proteomic changes during the embryonic period are not well characterized. We analyzed the proteomic alterations over the complete time course of honey bee worker embryogenesis at 24, 48, and 72 h of age, using mass spectrometry-based proteomics, label-free quantitation, and bioinformatics. Of the 1460 proteins identified the embryo of all three ages, the core proteome (proteins shared by the embryos of all three ages, accounting for 40%) was mainly involved in protein synthesis, metabolic energy, development, and molecular transporter, which indicates their centrality in driving embryogenesis. However, embryos at different developmental stages have their own specific proteome and pathway signatures to coordinate and modulate developmental events. The young embryos (<24 h) stronger expression of proteins related to nutrition storage and nucleic acid metabolism may correlate with the cell proliferation occurring at this stage. The middle aged embryos (24 -48 h) enhanced expression of proteins associated with cell cycle control, transporters, antioxidant activity, and the cytoskeleton suggest their roles to support rudimentary organogenesis. Among these proteins, the biological pathways of aminoacyl-tRNA biosynthesis, ␤-alanine metabolism, and protein export are intensively activated in the embryos of middle age. The old embryos (48 -72 h) elevated expression of proteins implicated in fatty acid metabolism and morphogenesis indicate their functionality for the formation and development of organs and dorsal closure, in which the biological pathways of fatty acid metabolism and RNA transport are highly activated. These findings add novel understanding to the molecular details of honey bee embryogenesis, in which the programmed activation of the proteome matches with the physiological transition observed during embryogenesis. The identified biological pathways and key node proteins allow for further functional analysis and genetic manipulation for both the honey bee embryos and other eusocial insects. Molecular & Cellular Proteomics 13:

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Section: Ligustica) Embryos (Normalized By Gapdh) (A) Is Significantsupporting

confidence: 69%

In-depth Proteomics Characterization of Embryogenesis of the Honey Bee Worker (Apis mellifera ligustica)

Mao

Han

et al. 2014

Molecular & Cellular Proteomics

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“…Such profiles were found for mrjp3, mrjp5, apisimin and regucalcin, which all encode LJ proteins. The profile was known for mrjp3 (Feng et al 2009) but it was novel for the other genes. These genes can be potentially examined as candidate reference genes for genetic (epigenetic) expression studies of genes encoding other LJ proteins possessing the same transcription profile in nurse head.…”

Section: Discussionmentioning

confidence: 99%

“…Amylase is abundantly expressed in foragers and very weakly in nurses (Ohashi et al 1999;Liu et al 2011) and it was selected as a marker for foragers. Gapdh, actin and cyclophilin were selected as commonly used reference genes (Scharlaken et al 2008;Lourenço et al 2008;Pfaffl 2004), whereas psa1 due to its presumed stable expression in workers of various age (Feng et al 2009). Primers with higher Tm (60-70…”

Section: Honeybee Samplesmentioning

confidence: 99%

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Identification and validation of reference genes for real time PCR expression studies in heads of nurse honeybees

et al. 2013

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Abstract:The primary aim of this study was to identify reference genes and workers of particular role and ages that would be suitable for exploring genetic/epigenetic variations in constitutive expression of a gene encoding antimicrobial peptide defensin1 in worker heads using real-time PCR. This peptide is an integral component of larval food and honey and has potential to act against some brood pathogens. Expression levels of distinct genes may vary in worker heads due to genetic factors, age of bee, and particular role of a worker that depends on its age or colony needs. Prerequisite for exploring the variations in defensin1 expression was therefore to identify such workers in which correlated expression of defensin1 and suitable reference genes occurs. Selection process was done by carefully designed quantitative real-time PCR procedure in two colonies showing different age-related division of labor. Expression of ten candidate reference genes, defensin1 and amylase, as a marker of forager bees, was assessed in pooled head samples of workers aged 2 to 30 days. Correlated and moreover stable expression of defensin1 and six candidate genes was detected in nursing bees in both colonies. The suitable reference genes were therefore selected on the basis of their expression stability. This was evaluated by geNorm and NormFinder algorithms in pooled head samples and through plotted Cq data in head samples of individual nurse bees. As the best reference genes were selected: psa1, tctp1, cyclophilin, gapdh and mrjp4 (in this order). They are suitable for aforementioned defensin1 expression studies and also for studies of other genes expressed in heads of nurses. In addition, an amylase expression-based procedure for reliable distinguishing nurses from foragers was elaborated.

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“…We believe that the MRJP genes might be similarly regulated as defensin1 gene during aging of nurses. This is important because age-dependent development of hypopharyngeal glands in nurses was shown to be associated with changes in protein synthesis (Knecht & Kaatz 1990;Ohashi et al 1997;Feng et al 2009). Similar profiles of relative defensin1 mRNA levels in nurse heads were obtained with both reference mRNAs.…”

Section: Discussionmentioning

confidence: 99%

Expression of larval jelly antimicrobial peptide defensin1 in Apis mellifera colonies

et al. 2012

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Honeybee brood food, larval jelly (LJ) contains antimicrobial peptide defensin1 that is able to inhibit in vitro growth of the pathogen causing American foulbrood (AFB). This fact suggests that LJ defensin1 could participate in defense of colonies against AFB. We assume that the potential defense function of defensin1 in vivo might depend on its amount in LJs. Therefore, we investigated the expression of defensin1 in colonies. The expression was examined on protein and mRNA levels in colonies of several Apis mellifera carnica lines collected in 3 apiaries (1 infected with AFB) with the aim to identify factors influencing the expression. Levels of defensin1 were determined in royal and worker jellies by a developed immunoblot procedure employing antibodies generated against the recombinant peptide. Defensin1 mRNA levels in nurse heads were explored by dot blot hybridization using transcript of two MRJP genes for normalization. Analyzed LJs contained various amounts of defensin1 (0.159-0.524 µg/mg jelly). Higher variations in defensin1 levels were observed among LJ samples collected from different colonies than among those collected within single colony. Colonies producing LJs with elevated defensin1 levels occurred among various honeybee lines. Levels of defensin1 mRNA varied in heads of nurses and the variations correlated with defensin1 peptide levels in LJs only in some colonies. Obtained data demonstrate that defensin1 is constitutively expressed into LJs in colonies and indicate that its levels in jellies are determined by genetic factors regulating transcription and/or translation/posttranslation processes in nurses. AFB infection, larval age and type of LJ do not seem to affect the levels of the peptide in LJs. Findings made in this work suggest that it should be possible to breed novel honeybee lines expressing higher amounts of defensin1 into LJs.

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Proteomic analysis of honeybee worker (Apis mellifera) hypopharyngeal gland development

Cited by 76 publications

References 54 publications

In-depth Proteomics Characterization of Embryogenesis of the Honey Bee Worker (Apis mellifera ligustica)

In-depth Proteomics Characterization of Embryogenesis of the Honey Bee Worker (Apis mellifera ligustica)

Identification and validation of reference genes for real time PCR expression studies in heads of nurse honeybees

Expression of larval jelly antimicrobial peptide defensin1 in Apis mellifera colonies

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