Expression of Two Ecdysteroid-Regulated Genes, Broad-Complex and E75, in the Brain and Ovary of the Honeybee (Apis mellifera L.)

Abstract We previously demonstrated that two ecdysteroid-regulated genes, Mblk-1/E93 and E74, are expressed selectively in Kenyon cell subtypes in the mushroom bodies of the honeybee (Apis mellifera L.) brain. To further examine the possible involvement of ecdysteroid-regulated genes in brain function as well as in oogenesis in the honeybee, we isolated cDNAs for two other ecdysteroid-regulated genes, Broad-Complex (BR-C) and E75, and analyzed their expression in the worker brain as well as in the queen abdomen. In situ hybridization revealed that BR-C, like Mblk-1/E93, is expressed selectively in the large-type Kenyon cells of the mushroom bodies in the worker brain, whereas E75 is expressed in all mushroom body neuron subtypes, suggesting a difference in the mode of response to ecdysteroid among Kenyon cell subtypes. In the queen ovary, both BR-C and E75 are expressed preferentially in the follicle cells that surround egg cells at the late stage, suggesting their role in oogenesis. These results suggest that BR-C and E75 are involved in the regulation of brain function as well as in reproductive physiology in the adult honeybee.

[1]  K. Hartfelder,et al.  Caste and metamorphosis: hemolymph titers of juvenile hormone and ecdysteroids in last instar honeybee larvae. , 1990, General and comparative endocrinology.

[2]  C. Thummel,et al.  Loss of the ecdysteroid-inducible E75A orphan nuclear receptor uncouples molting from metamorphosis in Drosophila. , 2002, Developmental cell.

[3]  M. Bownes,et al.  Translating Available Food Into the Number of Eggs Laid by Drosophila melanogaster , 2004, Genetics.

[4]  J. Fristrom,et al.  A switch in broad-complex zinc-finger isoform expression is regulated posttranscriptionally during the metamorphosis of Drosophila imaginal discs. , 1996, Developmental biology.

[5]  P. Sharp,et al.  Splicing of messenger RNA precursors. , 1987, Science.

[6]  H. Takeuchi,et al.  DNA-binding properties of Mblk-1, a putative transcription factor from the honeybee. , 2002, Biochemical and biophysical research communications.

[7]  A. Raikhel,et al.  Mosquito ecdysteroid receptor: analysis of the cDNA and expression during vitellogenesis. , 1995, Insect biochemistry and molecular biology.

[8]  S. Takiya,et al.  Structure and expression of the gene encoding a Broad-Complex homolog in the silkworm, Bombyx mori. , 2004, Gene.

[9]  M. Heisenberg What do the mushroom bodies do for the insect brain? an introduction. , 1998, Learning & memory.

[10]  P. Mobbs The Brain of the Honeybee Apis Mellifera. I. The Connections and Spatial Organization of the Mushroom Bodies , 1982 .

[11]  P. Sharp,et al.  Splicing of messenger RNA precursors. , 1987, Annual Review of Biochemistry.

[12]  H. Takeuchi,et al.  Preferential expression of the gene for a putative inositol 1,4,5-trisphosphate receptor homologue in the mushroom bodies of the brain of the worker honeybee Apis mellifera L. , 1998, Biochemical and biophysical research communications.

[13]  K. Iatrou,et al.  The orphan nuclear receptors BmE75A and BmE75C of the silkmoth Bombyx mori: hornmonal control and ovarian expression. , 2002, Insect biochemistry and molecular biology.

[14]  M. Bownes,et al.  The function of the broad-complex during Drosophila melanogaster oogenesis. , 1999, Genetics.

[15]  L. Margaritis,et al.  Dynamics of apoptosis in the ovarian follicle cells during the late stages of Drosophila oogenesis , 2002, Cell and Tissue Research.

[16]  A. Kamikouchi,et al.  Concentrated expression of Ca2+/calmodulin‐dependent protein kinase II and protein kinase C in the mushroom bodies of the brain of the honeybee Apis mellifera L. , 2000, The Journal of comparative neurology.

[17]  R. Menzel,et al.  Integrative properties of the Pe1 neuron, a unique mushroom body output neuron. , 1998, Learning & memory.

[18]  R. Levine,et al.  The Steroid Hormone 20-Hydroxyecdysone Enhances Neurite Growth ofDrosophila Mushroom Body Neurons Isolated during Metamorphosis , 1998, The Journal of Neuroscience.

[19]  J. A. Svoboda,et al.  Makisterone A: The major ecdysteroid from the pupa of the honey bee, Apis mellifera , 1985 .

[20]  D. Withers,et al.  The Drosophila Broad-Complex encodes a family of related proteins containing zinc fingers. , 1991, Genetics.

[21]  T. Kadowaki,et al.  The Changes of Gene Expression in Honeybee (Apis mellifera) Brains Associated with Ages , 2004, Zoological science.

[22]  K. Hartfelder,et al.  Ecdysteroid titer and reproduction in queens and workers of the honey bee and of a stingless bee: loss of ecdysteroid function at increasing levels of sociality? , 2002, Insect biochemistry and molecular biology.

[23]  G. Robinson,et al.  Comparison of juvenile hormone and ecdysteroid haemolymph titres in adult worker and queen honey bees (Apis mellifera) , 1991 .

[24]  L. Restifo,et al.  Mutations in a steroid hormone-regulated gene disrupt the metamorphosis of the central nervous system in Drosophila. , 1991, Developmental biology.

[25]  A. Kamikouchi,et al.  Identification of a novel gene, Mblk‐1, that encodes a putative transcription factor expressed preferentially in the large‐type Kenyon cells of the honeybee brain , 2001, Insect molecular biology.

[26]  M. Bownes,et al.  Two signalling pathways specify localised expression of the Broad-Complex in Drosophila eggshell patterning and morphogenesis. , 1997, Development.

[27]  A. Kamikouchi,et al.  Identification and punctate nuclear localization of a novel noncoding RNA, Ks-1, from the honeybee brain. , 2002, RNA.

[28]  Y. Fujiwara,et al.  cDNA cloning, gene structure, and expression of Broad-Complex (BR-C) genes in the silkworm, Bombyx mori. , 2004, Insect biochemistry and molecular biology.

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

[30]  M. Mckeown,et al.  Drosophila ultraspiracle modulates ecdysone receptor function via heterodimer formation , 1992, Cell.

[31]  K. Hartfelder,et al.  Ecdysteroid titers in pupae of highly social bees relate to distinct modes of caste development. , 2002, Journal of insect physiology.

[32]  C. Thummel,et al.  The Drosophila E93 gene from the 93F early puff displays stage- and tissue-specific regulation by 20-hydroxyecdysone. , 1995, Developmental biology.

[33]  L. Cooley,et al.  Ecdysone response genes govern egg chamber development during mid-oogenesis in Drosophila. , 1999, Development.

[34]  Guoqiang Sun,et al.  Two isoforms of the early E74 gene, an Ets transcription factor homologue, are implicated in the ecdysteroid hierarchy governing vitellogenesis of the mosquito, Aedes aegypti , 2002, Molecular and Cellular Endocrinology.

[35]  E. Berger,et al.  Juvenile hormone regulation of the E75 nuclear receptor is conserved in Diptera and Lepidoptera. , 2004, Gene.

[36]  L. Riddiford,et al.  Juvenile hormone prevents ecdysteroid-induced expression of broad complex RNAs in the epidermis of the tobacco hornworm, Manduca sexta. , 1998, Developmental biology.

[37]  Prepro-tachykinin gene expression in the brain of the honeybee Apis mellifera , 2004, Cell and Tissue Research.

[38]  M. Winston,et al.  Insect societies and the molecular biology of social behavior , 1997, BioEssays : news and reviews in molecular, cellular and developmental biology.

[39]  C. Thummel,et al.  E93 directs steroid-triggered programmed cell death in Drosophila. , 2000, Molecular cell.

[40]  Gene expression of ecdysteroid‐regulated gene E74 of the honeybee in ovary and brain , 2005, Insect molecular biology.

[41]  Tzumin Lee,et al.  TGF-β Signaling Activates Steroid Hormone Receptor Expression during Neuronal Remodeling in the Drosophila Brain , 2003, Cell.

[42]  G. Robinson,et al.  Selective neuroanatomical plasticity and division of labour in the honeybee , 1993, Nature.

[43]  M. Bownes,et al.  Nutritional status affects 20-hydroxyecdysone concentration and progression of oogenesis in Drosophila melanogaster. , 2005, The Journal of endocrinology.

[44]  E. Baehrecke,et al.  Caspases function in autophagic programmed cell death in Drosophila , 2003, Development.

[45]  N. Strausfeld Organization of the honey bee mushroom body: Representation of the calyx within the vertical and gamma lobes , 2002, The Journal of comparative neurology.

[46]  L. Margaritis,et al.  Programmed cell death of follicular epithelium during the late developmental stages of oogenesis in the fruit flies Bactrocera oleae and Ceratitis capitata (Diptera, Tephritidae) is mediated by autophagy , 2006, Development, growth & differentiation.

[47]  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.

[48]  Peri T Kurshan,et al.  Developmental changes in expression patterns of two dopamine receptor genes in mushroom bodies of the honeybee, Apis mellifera , 2003, The Journal of comparative neurology.

[49]  D. Hogness,et al.  The E75 ecdysone-inducible gene responsible for the 75B early puff in Drosophila encodes two new members of the steroid receptor superfamily. , 1990, Genes & development.

[50]  M. Bownes,et al.  E75A and E75B have opposite effects on the apoptosis/development choice of the Drosophila egg chamber , 2006, Cell Death and Differentiation.

[51]  C. Thummel,et al.  Steroid Regulation of Postembryonic Development and Reproduction in Drosophila , 2000, Trends in Endocrinology & Metabolism.

[52]  Ronald L. Davis Mushroom bodies and drosophila learning , 1993, Neuron.

[53]  A. Raikhel,et al.  The early gene Broad is involved in the ecdysteroid hierarchy governing vitellogenesis of the mosquito Aedes aegypti. , 2004, Journal of molecular endocrinology.