Next‐generation small RNA sequencing for microRNAs profiling in Apis mellifera: comparison between nurses and foragers

MicroRNAs (miRNAs) are endogenous small non‐coding RNAs regulating gene expression in animals and plants. To find some differentially expressed miRNAs that may be associated with age‐dependent behavioural changes in honey bees (Apis mellifera), we applied next‐generation high‐throughput sequencing technology to detect small RNAs in nurses and foragers. Our results showed that both nurses and foragers had a complicated small RNA population, and the length of small RNAs varied, 22 nucleotides being the predominant length. Combining deep sequencing and bioinformatic analysis, we discovered that nine known miRNAs were significantly different between nurses and foragers (P < 0.01; absolute value of fold‐change ≥1). Some of their target genes were related to neural function. Moreover, 67 novel miRNAs were identified in nurses and foragers. Ame‐miR‐31a and ame‐miR‐13b were further validated using quantitative reverse‐transcription PCR assays. The present study provides new information on the miRNA abundance of honey bees, and enhances our understanding of miRNA function in the regulation of honey bee development.

[1]  S. Su,et al.  High-abundance mRNAs in Apis mellifera: comparison between nurses and foragers. , 2011, Journal of insect physiology.

[2]  Songnian Hu,et al.  Next‐generation small RNA sequencing for microRNAs profiling in the honey bee Apis mellifera , 2010, Insect molecular biology.

[3]  Jun Wang,et al.  Genomic Comparison of the Ants Camponotus floridanus and Harpegnathos saltator , 2010, Science.

[4]  C. Whitfield,et al.  Correlated expression patterns of microRNA genes with age‐dependent behavioural changes in honeybee , 2010, Insect molecular biology.

[5]  V. Beneš,et al.  The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. , 2009, Clinical chemistry.

[6]  X. Chen,et al.  Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases , 2008, Cell Research.

[7]  Phillip D Zamore,et al.  Beginning to understand microRNA function , 2007, Cell Research.

[8]  Christine G Elsik,et al.  Computational and transcriptional evidence for microRNAs in the honey bee genome , 2007, Genome Biology.

[9]  Xianwu Zheng,et al.  Role of Arabidopsis AGO6 in siRNA accumulation, DNA methylation and transcriptional gene silencing , 2007, The EMBO journal.

[10]  Kunio Inoue.  [MicroRNA function in animal development]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[11]  Ying Wang,et al.  Insights into social insects from the genome of the honeybee Apis mellifera , 2006, Nature.

[12]  K. Livak,et al.  Real-time quantification of microRNAs by stem–loop RT–PCR , 2005, Nucleic acids research.

[13]  G. Robinson,et al.  Behavior and the limits of genomic plasticity: power and replicability in microarray analysis of honeybee brains , 2005, Genes, brain, and behavior.

[14]  G. Robinson,et al.  Sociogenomics: social life in molecular terms , 2005, Nature Reviews Genetics.

[15]  R. Huber.  Amines and motivated behaviors: a simpler systems approach to complex behavioral phenomena , 2005, Journal of Comparative Physiology A.

[16]  G. Lenaers,et al.  Apisα2, Apisα7-1 and Apisα7-2: three new neuronal nicotinic acetylcholine receptor α-subunits in the honeybee brain , 2005 .

[17]  G. Gisselmann,et al.  Functional characterization of I h‐channel splice variants from Apis mellifera , 2004, FEBS letters.

[18]  G. Robinson,et al.  Phenotypic deconstruction reveals involvement of manganese transporter malvolio in honey bee division of labor , 2004, Journal of Experimental Biology.

[19]  A. Kamikouchi,et al.  Identification of Honeybee Antennal Proteins/Genes Expressed in a Sex- and/or Caste Selective Manner , 2004, Zoological science.

[20]  Ruedi Aebersold,et al.  Identification of androgen-coregulated protein networks from the microsomes of human prostate cancer cells , 2003, Genome Biology.

[21]  Gene E Robinson,et al.  Pheromone-mediated gene expression in the honey bee brain , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[22]  G. Robinson,et al.  Gene Expression Profiles in the Brain Predict Behavior in Individual Honey Bees , 2003, Science.

[23]  T. Kubo,et al.  The Activity of Mblk-1, a Mushroom Body-selective Transcription Factor from the Honeybee, Is Modulated by the Ras/MAPK Pathway* , 2003, The Journal of Biological Chemistry.

[24]  C. Burge,et al.  Vertebrate MicroRNA Genes , 2003, Science.

[25]  Xiaofeng Cao,et al.  ARGONAUTE4 Control of Locus-Specific siRNA Accumulation and DNA and Histone Methylation , 2003, Science.

[26]  Gene E Robinson,et al.  Genomics and Integrative Analyses of Division of Labor in Honeybee Colonies , 2002, The American Naturalist.

[27]  P. Ebert,et al.  Characterization of a chemosensory protein (ASP3c) from honeybee (Apis mellifera L.) as a brood pheromone carrier. , 2002, European journal of biochemistry.

[28]  Julia Krüger,et al.  Influence of Gene Action Across Different Time Scales on Behavior , 2002, Science.

[29]  R. Kucharski,et al.  Evaluation of differential gene expression during behavioral development in the honeybee using microarrays and northern blots , 2002, Genome Biology.

[30]  Shenglu Chen,et al.  An introduction to high-yielding royal jelly production methods in China , 2002 .

[31]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[32]  R. Menzel.  Searching for the memory trace in a mini-brain, the honeybee. , 2001, Learning & memory.

[33]  U. Müller.  Prolonged Activation of cAMP-Dependent Protein Kinase during Conditioning Induces Long-Term Memory in Honeybees , 2000, Neuron.

[34]  G. Robinson,et al.  Biogenic amines and division of labor in honey bee colonies , 1999, Journal of Comparative Physiology A.

[35]  P. Ebert,et al.  Isolation of seven unique biogenic amine receptor clones from the honey bee by library scanning , 1998, Insect molecular biology.

[36]  J. Claverie,et al.  The significance of digital gene expression profiles. , 1997, Genome research.

[37]  G. Lenaers,et al.  Apisalpha2, Apisalpha7-1 and Apisalpha7-2: three new neuronal nicotinic acetylcholine receptor alpha-subunits in the honeybee brain. , 2005, Gene.

[38]  Anton J. Enright,et al.  MicroRNA Targets in Drosophila , 2003, Genome Biology.

[39]  Thomas D. Schmittgen,et al.  Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2 2 DD C T Method , 2022 .