DOCSLIB.ORG

Abnormalities of Male-Specific FRU Protein and Serotonin Expression

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Abnormalities of Male-Specific FRU Protein and Serotonin Expression The Journal of Neuroscience, January 15, 2001, 21(2):513–526 Abnormalities of Male-Specific FRU Protein and Serotonin Expression in the CNS of fruitless Mutants in Drosophila Gyunghee Lee and Jeffrey C. Hall Department of Biology, Brandeis University, Waltham, Massachusetts 02454 The fruitless gene in Drosophila produces male-specific protein include courtship among males, which has been hypothesized (FRU M) involved in the control of courtship. FRU M spatial and to involve anomalies of serotonin (5-HT) function in the brain. temporal patterns were examined in fru mutants that exhibit However, double-labeling uncovered no coexpression of FRU M aberrant male courtship. Chromosome breakpoints at the locus and 5-HT in brain neurons. Yet, a newly identified set of sexually eliminated FRU M. Homozygous viable mutants exhibited an dimorphic FRU M/5-HT-positive neurons was identified in the intriguing array of defects. In fru 1 males, there were absences abdominal ganglion of adult males. These sexually dimorphic of FRU M-expressing neuronal clusters or stained cells within neurons (s-Abg) project toward regions of the abdomen in- certain clusters, reductions of signal intensities in others, and volved in male reproduction. The s-Abg neurons and the prox- ectopic FRU M expression in novel cells. fru 2 males exhibited an imal extents of their axons were unstained or absent in wild-type overall decrement of FRU M expression in all neurons normally females and exhibited subnormal or no 5-HT immunoreactivity in expressing the gene. fru 4 and fru sat mutants only produced certain fru-mutant males, indicating that fruitless controls the for- FRU M in small numbers of neurons at extremely low levels, and mation of these cells or 5-HT production in them. no FRU M signals were detected in fru 3 males. This array of Key words: fruitless transposons; chromosome aberrations; abnormalities was inferred to correlate with the varying behav- brain neurons; ventral nerve cord; sexual dimorphism; seroto- ioral defects exhibited by these mutants. Such abnormalities nergic neurons Courtship in Drosophila melanogaster is regulated by a somatic behavior caused by fru mutations could be understood in terms of sex-determination hierarchy. One of the “downstream” genes where FRU M is expressed in the CNS (or not expressed, as the functioning within this hierarchy is fruitless (for review, see case may be) in a given mutant. Goodwin, 1999; Yamamoto and Nakano, 1999). fru mutations The fact that several fru-mutant types court other males might cause the most sharply defined effects on male courtship, com- be attributable to subnormal levels of serotonin (5-HT) in rele- pared with behaviorally mutant phenotypes associated with other vant brain cells. This hypothesis suggested itself because of the downstream genes (Villella and Hall, 1996; Finley et al., 1997). anomalous inter-male courtships that are induced by ectopic ϩ fruitless produces male- and female-specific transcripts under the expression of the white (w ) gene (Zhang and Odenwald, 1995; control of a distal promoter (P1) located ϳ100 kb from the bulk Hing and Carlson, 1996). white encodes a tryptophan–guanine of the open reading frame of the gene (Ryner et al., 1996). The transporter; because tryptophan is a precursor of 5-HT, induced M ϩ male-specific proteins (FRU s) encoded by P1-controlled mR- w expression all over the brain could cause subnormal 5-HT NAs are likely to be involved in the regulation of courtship or the levels in neurons that normally produce it. Drug-induced 5-HT development of the neural substrates for male reproductive be- reductions can induce homosexual behavior of male mammals havior (Goodwin 1999). In females, P1-produced transcripts are (for review, see Gessa and Tagliamonte, 1974; Fratta et al., 1977). not translated into detectable FRU protein (Lee et al., 2000; fru mutants, which are predicted to exhibit deficits in male- Usui-Aoki et al., 2000). specific transcription factors encoded by this gene (Goodwin, One approach toward understanding how fru regulates male 1999; Goodwin et al., 2000), could also be deficient in 5-HT if its M courtship is to compare patterns of FRU expression in the CNS production is downstream of FRU M function. Moreover, the ϩ of various fruitless mutants that display behavioral phenotypes actions of both ectopic-w and fruitless have been suggested to be ranging from mildly to severely defective (Villella et al., 1997; in the same pathway because of a blockade of the behavioral effects ϩ Goodwin et al., 2000). The courtship subnormalities and bisexual of ectopic-w by a frumutation that, by itself, causes very low levels Received May 18, 2000; revised Oct. 3, 2000; accepted Oct. 30, 2000. of male courtship (Nilsson et al., 2000). Based on these suppositions, This work was supported by National Institutes of Health (NIH) Grants NS-33352 we analyzed the relationship between the spatial distribution of and GM-21473, and NIH Neuroscience Training Grant 5T32-NS-07292. We thank 5-HT and fru gene products in normal and mutant CNSs. Jae H. Park, Adriana Villella, Stephen F. Goodwin, Jean-Christophe Billeter, Bruce Along with demonstrating that elements of 5-HT production S. Baker, and Barbara J. Taylor for helpful discussions, and we appreciate comments on this manuscript from Barbara J. Taylor, Margit Foss, and Ralph J. Greenspan. We are downstream of fru functioning (although not in CNS regions thank Edward Dougherty and Maki Kaneko for confocal and photographic assis- that one might have expected), this feature of the study provided tance, and Barbara Beltz and Kalpana White for advice about anti-5-HT M immunohistochemistry. the first information on axonal projections of certain FRU cells. Correspondence should be addressed to Jeffrey C. Hall, Department of Biology, Assessing the subcellular localization of FRU M alone gives no MS-008, Brandeis University, Waltham, MA 02454-9110. E-mail: [email protected]. insight into this matter; FRU immunoreactivities are nuclear Dr. Lee’s present address: Department of Biochemistry and Cellular and Molec- ular Biology, University of Tennessee, Knoxville, TN 37996-0840. (Lee et al., 2000), consistent with the supposition that these Copyright © 2001 Society for Neuroscience 0270-6474/01/210513-14$15.00/0 proteins are gene regulators. 514 J. Neurosci., January 15, 2001, 21(2):513–526 Lee and Hall • FRUM and 5-HT Abnormalities in fruitless Courtship Mutants MATERIALS AND METHODS CNSs dissected from 1-d-old Df-fru 4–40/Df-sat 15 or Df-Cha M5/Df-sat 15 (double-deletion) male pupae. The particular probe (called P1.S1; see Strains and culturing. Stocks and progeny from crosses of D. melanogaster Fig. 1) and the labeling procedures were as described in Lee et al. (2000). were reared in 12 hr light/dark (LD) cycles at 25°C, 70% relative Scoring of staining intensities. To analyze the levels of FRU M in CNSs humidity, on a sucrose–cornmeal–yeast medium containing the mold of viable fru mutants and wild-type males, fluorescently immunostained inhibitor Tegosept. A Canton-S strain was used as the wild-type control. signals from various FRU M-expressing neuronal clusters in whole- The fruitless mutant stocks fru1, fru2, fru3, fru4, and frusat were mounted CNS were quantified as described in Lee et al. (2000). We maintained as described in Villella et al. (1997) and Goodwin et al. focused on three such clusters within the superior protocerebrum in (2000). The following homozygous-lethal fru variants, missing all or analyzing the wild type and the fru mutants. These and other FRU M cell part of the locus, were combined in pairwise crosses (see Table 1): M5 sat15 w24 groups had staining intensities assessed for animals carrying the various Df(3R)Cha , Df(3R)P14, Df(3R)fru , Df(3R)fru , and fru genotypes (see Table 2 legend). The FRU M immunostaining quan- Df(3R)fru 4–40 (Gailey and Hall, 1989; Ito et al., 1996; Ryner et al., 1996; M5 tifications were performed on whole-mounted brains dissected from Anand et al., 2001). These deletions will be referred to as Df-Cha , 2-d-old pupae, which show the highest level of CNS expression by this Df-P14, Df-sat 15, Df-fru w24, and Df-fru 4–40. The lethal variants fruw12 and w27 protein (Lee et al., 2000), and from 4- to 7-d-old adults, which are active fru , each carrying single breakpoints within the locus, were crossed to courters. The specimens to be compared (in terms of genotype or each other or to certain Df-bearing flies to generate severely lesioned life-cycle stage) were processed simultaneously to minimize signal vari- genotypes (see Table 1). The Df-fru or fru-lethal stocks were balanced with ations that could occur for nonsubstantive reasons. At least five CNSs In(3LR)TM6B, Tb, In(3LR)TM3, Sb,orTp(3)MKRS, Sb. were sampled from animals of each genotype at both of the different To obtain 2-d-old pupae homozygous for a fru mutation or carrying a stages. The dorsoanterior brain region that contains both neuronal clus- given transheterozygous combination, flies from TM6B-balanced stocks ters was imaged at 100ϫ magnification by confocal microscopy (2 or 4 ␮m were crossed to each other. This was necessary because of the homozy- optical sections). Staining intensities for nuclei of cells within these two gous lethality or sterility associated with fru variants, with the exception clusters were obtained (as pixel values) from at least five individual 2 of fru , which is homozygous-viable and fertile (permitting pupae to be brains by applying an Adobe PhotoShop (3.0) tool called Histogram. obtained from a true-breeding stock). For the other strains, balancer- This permitted an average value to be computed from several strongly over-fru heterozygotes were crossed, and animals not expressing the stained nuclei in the cells of the two nearby clusters; for a given brain, pupal marker Tb were selected.
Load more

Recommended publications
  • Fruitless Splicing Specifies Male Courtship Behavior in Drosophila
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Cell, Vol. 121, 785–794, June 3, 2005, Copyright ©2005 by Elsevier Inc. DOI 10.1016/j.cell.2005.04.027 fruitless Splicing Specifies Male Courtship Behavior in Drosophila Ebru Demir and Barry J. Dickson* sexual behaviors. These behaviors are essential for Institute of Molecular Biotechnology of the Austrian their reproductive success, and so strong selective Academy of Sciences pressure is likely to have favored the evolution of genes Dr. Bohr-Gasse 3–5 that “hardwire” them into the brain. The initial steps of A-1030 Vienna sexual differentiation have been well characterized for Austria several model organisms, and genetic perturbations in these sex-determination hierarchies can alter all as- pects of the sexual phenotype—innate behaviors as Summary well as gross anatomy. Several genes near the top of these sex-determination hierarchies thus qualify as de- All animals exhibit innate behaviors that are specified velopmental switch genes, but they cannot be consid- during their development. Drosophila melanogaster ered specifically as behavioral switch genes. A switch males (but not females) perform an elaborate and in- gene for a sexual behavior should act to specify either nate courtship ritual directed toward females (but not male or female behavior, irrespective of the overall sex- males). Male courtship requires products of the fruit- ual phenotype of the animal. A candidate for such a less (fru) gene, which is spliced differently in males gene is the fruitless (fru) gene of Drosophila, which is and females.
    [Show full text]
  • Behavior and Cytogenetics of Fruitless in Drosophila Melanogaster
    Copyright 0 1989 by the Genetics Society of Amerlcd Behavior and Cytogenetics offruitless in Drosophila melanogaster: Different Courtship Defects Caused by Separate, Closely Linked Lesions Donald A. Gailey and JeffreyC. Hall Department of Biology, Brandeis University, Waltham, Massachusetts02254 Manuscript received September 22, 1988 Accepted for publication December 24, 1988 ABSTRACT Thefruitless (fru)courtship mutant was dissected into three defectsof male reproductive behavior, which were separable as to their genetic etiologies by application of existing and newly induced chromosomal aberrations.fru itself is a small inversion [In(3R)9OC; 9 1B] on genetic andcytological criteria. Uncovering the fru distal breakpoint with deletions usually led to males with two of thefru courtship abnormalities: no copulation attemptswith females (hence, behavioralsterility) and vigorous courtship among males, including the formation of “courtship chains.” However, certain genetic changes involving region 91B resulted in males who formed courtship chains but who mated with females. Uncovering the fru proximal breakpoint led to males that passively elicit inappropriately high levels of courtship. Thiselicitation property was separable genetically from thesterility and chain formation phenotypes and provisionally mapped to the interval 89F-90F, which includes the fru proximal breakpoint. Behavioral sterility and chaining werealso observed in males expressing certain abnormal genotypes, independent of thefru inversion. These included combinations ofdeficiencies,
    [Show full text]
  • Sexual Dimorphism in Diverse Metazoans Is Regulated by a Novel Class of Intertwined Zinc Fingers
    Downloaded from genesdev.cshlp.org on October 4, 2021 - Published by Cold Spring Harbor Laboratory Press Sexual dimorphism in diverse metazoans is regulated by a novel class of intertwined zinc fingers Lingyang Zhu,1,4 Jill Wilken,2 Nelson B. Phillips,3 Umadevi Narendra,3 Ging Chan,1 Stephen M. Stratton,2 Stephen B. Kent,2 and Michael A. Weiss1,3–5 1Center for Molecular Oncology, Departments of Biochemistry & Molecular Biology and Chemistry, The University of Chicago, Chicago, Illinois 60637-5419 USA; 2Gryphon Sciences, South San Francisco, California 94080 USA; 3Department of Biochemistry, Case Western Reserve School of Medicine, Cleveland, Ohio 44106-4935 USA Sex determination is regulated by diverse pathways. Although upstream signals vary, a cysteine-rich DNA-binding domain (the DM motif) is conserved within downstream transcription factors of Drosophila melanogaster (Doublesex) and Caenorhabditis elegans (MAB-3). Vertebrate DM genes have likewise been identified and, remarkably, are associated with human sex reversal (46, XY gonadal dysgenesis). Here we demonstrate that the structure of the Doublesex domain contains a novel zinc module and disordered tail. The module consists of intertwined CCHC and HCCC Zn2+-binding sites; the tail functions as a nascent recognition ␣-helix. Mutations in either Zn2+-binding site or tail can lead to an intersex phenotype. The motif binds in the DNA minor groove without sharp DNA bending. These molecular features, unusual among zinc fingers and zinc modules, underlie the organization of a Drosophila enhancer that integrates sex- and tissue-specific signals. The structure provides a foundation for analysis of DM mutations affecting sexual dimorphism and courtship behavior.
    [Show full text]
  • Fruitless Tunes Functional Flexibility of Courtship Circuitry During Development
    SHORT REPORT fruitless tunes functional flexibility of courtship circuitry during development Jie Chen1, Sihui Jin1, Dandan Chen1, Jie Cao1, Xiaoxiao Ji1, Qionglin Peng1*, Yufeng Pan1,2* 1The Key Laboratory of Developmental Genes and Human Disease, School of Life Science and Technology, Southeast University, Nanjing, China; 2Co-innovation Center of Neuroregeneration, Nantong University, Nantong, China Abstract Drosophila male courtship is controlled by the male-specific products of the fruitless M M (fru ) gene and its expressing neuronal circuitry. fru is considered a master gene that controls all M aspects of male courtship. By temporally and spatially manipulating fru expression, we found that M fru is required during a critical developmental period for innate courtship toward females, while its function during adulthood is involved in inhibiting male–male courtship. By altering or M eliminating fru expression, we generated males that are innately heterosexual, homosexual, bisexual, or without innate courtship but could acquire such behavior in an experience-dependent M manner. These findings show that fru is not absolutely necessary for courtship but is critical during development to build a sex circuitry with reduced flexibility and enhanced efficiency, and M provide a new view about how fru tunes functional flexibility of a sex circuitry instead of switching on its function as conventionally viewed. Introduction Drosophila male courtship is one of the best understood innate behaviors in terms of genetic and neuronal mechanisms (Dickson, 2008; Yamamoto and Koganezawa, 2013). It has been well estab- *For correspondence: [email protected] (QP); lished that the fruitless (fru) gene and its expressing neurons control most aspects of such innate [email protected] (YP) behavior (Ito et al., 1996; Manoli et al., 2005; Ryner et al., 1996; Stockinger et al., 2005).
    [Show full text]
  • Optogenetic Activation of the Fruitless-Labeled Circuitry in Drosophila Subobscura Males Induces Mating Motor Acts
    11662 • The Journal of Neuroscience, November 29, 2017 • 37(48):11662–11674 Behavioral/Cognitive Optogenetic Activation of the fruitless-Labeled Circuitry in Drosophila subobscura Males Induces Mating Motor Acts Ryoya Tanaka, Tomohiro Higuchi, Soh Kohatsu, Kosei Sato, and Daisuke Yamamoto Division of Neurogenetics, Tohoku University, Graduate School of Life Sciences, Sendai 980-8577, Japan Itremainsanenigmahowthenervoussystemofdifferentanimalspeciesproducesdifferentbehaviors.Westudiedtheneuralcircuitryfor mating behavior in Drosophila subobscura, a species that displays unique courtship actions not shared by other members of the genera including the genetic model D. melanogaster, in which the core courtship circuitry has been identified. We disrupted the D. subobscura fruitless(fru) gene,amasterregulatorforthecourtshipcircuitryformationinD.melanogaster,resultingincompletelossofmatingbehavior.We also generated frusoChrimV, which expresses the optogenetic activator Chrimson fused with a fluorescent marker under the native fru promoter. Thefru-labeledcircuitryinD.subobscuravisualizedbyfrusoChrimV revealeddifferencesbetweenfemalesandmales,optogeneticactivation of which in males induced mating behavior including attempted copulation. These findings provide a substrate for neurogenetic dissec- tion and manipulation of behavior in non-model animals, and will help to elucidate the neural basis for behavioral diversification. Key words: courtship; CRISPR/Cas9; Drosophila; fruitless Significance Statement How did behavioral specificity arise during
    [Show full text]
  • Investigating the Role of Fruitless in Behavioural Isolation Between Drosophila Melanogaster and Drosophila Simulans
    Western University Scholarship@Western Electronic Thesis and Dissertation Repository 8-22-2017 10:00 AM Investigating the Role of Fruitless in Behavioural Isolation Between Drosophila melanogaster and Drosophila simulans Jalina Bielaska Da Silva The Unviersity of Western Ontario Supervisor Amanda J. Moehring The University of Western Ontario Graduate Program in Biology A thesis submitted in partial fulfillment of the equirr ements for the degree in Master of Science © Jalina Bielaska Da Silva 2017 Follow this and additional works at: https://ir.lib.uwo.ca/etd Part of the Ecology and Evolutionary Biology Commons Recommended Citation Bielaska Da Silva, Jalina, "Investigating the Role of Fruitless in Behavioural Isolation Between Drosophila melanogaster and Drosophila simulans" (2017). Electronic Thesis and Dissertation Repository. 4779. https://ir.lib.uwo.ca/etd/4779 This Dissertation/Thesis is brought to you for free and open access by Scholarship@Western. It has been accepted for inclusion in Electronic Thesis and Dissertation Repository by an authorized administrator of Scholarship@Western. For more information, please contact [email protected]. Abstract Behavioural isolation is a prezygotic mechanism that is usually determined by female preference, such as seen with the rejection behaviour exhibited by Drosophila simulans females to D. melanogaster males. To confirm the role of a previously identified candidate gene fruitless (fru) in behavioural isolation, I proposed to disrupt fru expression in both D. melanogaster and D. simulans to allow for the generation of interspecies hybrids expressing only a species-specific allele of fru. A reciprocal hemizygosity test would then be used to confirm the role of fru in behavioural isolation.
    [Show full text]
  • Fruitless Recruits Two Antagonistic Chromatin Factors to Establish Single-Neuron Sexual Dimorphism
    Fruitless Recruits Two Antagonistic Chromatin Factors to Establish Single-Neuron Sexual Dimorphism Hiroki Ito,1 Kosei Sato,1 Masayuki Koganezawa,1,4 Manabu Ote,1,4 Ken Matsumoto,2 Chihiro Hama,3 and Daisuke Yamamoto1,* 1Division of Neurogenetics, Tohoku University Graduate School of Life Sciences, Sendai 980-8577, Japan 2Japan Advanced Institute of Science and Technology, Nomi, Ishikawa 923-1292, Japan 3Faculty of Life Sciences, Kyoto Sangyo University, Kyoto 603-8555, Japan 4These authors contributed equally to this work *Correspondence: [email protected] DOI 10.1016/j.cell.2012.04.025 SUMMARY for phenotypic modifiers of fru in the compound eye yielded several factors involved in transcription machineries, including The Drosophila fruitless (fru) gene encodes a set of Bonus (Bon), a homolog of the mammalian TIF1 family of tran- putative transcription factors that promote male scriptional cofactors. Here, we demonstrate that Bon physically sexual behavior by controlling the development of associates with Fru in vivo, and the Fru-Bon complex recruits sexually dimorphic neuronal circuitry. However, the a chromatin regulator, Heterochromatin protein 1a (HP1a) or mechanism whereby fru establishes the sexual fate Histone deacetylase 1 (HDAC1), to discrete chromosomal sites. of neurons remains enigmatic. Here, we show that A reduction in HDAC1 activity enhanced the demasculinizing activity of fru mutations on the mAL neurons, whereas a reduction Fru forms a complex with the transcriptional cofactor in HP1a activity counteracted it. We propose that Fru bound to Bonus (Bon), which, in turn, recruits either of its target sites alters the chromatin configuration in collaboration two chromatin regulators, Histone deacetylase 1 with Bon, HP1a, and HDAC1, thereby orchestrating transcription (HDAC1), which masculinizes individual sexually of a series of downstream genes that function to determine the dimorphic neurons, or Heterochromatin protein 1a development of gender-typical neural circuitry.
    [Show full text]
  • Profile of Jeffrey C. Hall Hen Geneticist Jeffrey C
    PROFILE Profile of Jeffrey C. Hall hen geneticist Jeffrey C. Hall looks back on his ca- reer path, he offers praise for each of the personali- Wties he has encountered, from his deaf undergraduate advisor to his fellow Civil War scholars—not to mention the small, elegant, and urbane fruit fly. ‘‘They’re very complex organisms, quite sophisti- cated and interesting,’’ he says of Dro- sophila, the neurobiology and behavior of which have occupied his research since college. To Hall, Drosophila are valuable for more than just their easily manipulated genomes. As he grew to ‘‘love the fly,’’ he also began to delve into the insect’s genetics, neurobiology, and behavior at a deep level. Convinced of the worth of analyzing Drosophila mutants for study- ing behavior, Hall has dedicated his career to probing the neurobiological underpinnings of the fly’s courtship and behavioral rhythms. His research with Jeffrey C. Hall Drosophila genetics has elucidated the mechanisms of biological clocks and opened a window into the basis for fruit flies. Ives, who had been a student sophila laboratory of Lawrence Sandler, sexual differentiation in the nervous of geneticist Albert Sturtevant, lost his also a research descendent of Stur- system. hearing in graduate school and was rele- tevant’s. Hall worked with Sandler on Hall, a Professor of Biology at Bran- gated to a non-tenure-track research several research projects, starting with deis University (Waltham, MA), was a career that included shepherding under- the analysis of age-dependent enzyme recipient of the Genetics Society of graduate students like Hall through changes in Drosophila (2) but with a America Medal in 2003, the same year small projects.
    [Show full text]
  • Fruitless Splicing Specifies Male Courtship Behavior in Drosophila
    Cell, Vol. 121, 785–794, June 3, 2005, Copyright ©2005 by Elsevier Inc. DOI 10.1016/j.cell.2005.04.027 fruitless Splicing Specifies Male Courtship Behavior in Drosophila Ebru Demir and Barry J. Dickson* sexual behaviors. These behaviors are essential for Institute of Molecular Biotechnology of the Austrian their reproductive success, and so strong selective Academy of Sciences pressure is likely to have favored the evolution of genes Dr. Bohr-Gasse 3–5 that “hardwire” them into the brain. The initial steps of A-1030 Vienna sexual differentiation have been well characterized for Austria several model organisms, and genetic perturbations in these sex-determination hierarchies can alter all as- pects of the sexual phenotype—innate behaviors as Summary well as gross anatomy. Several genes near the top of these sex-determination hierarchies thus qualify as de- All animals exhibit innate behaviors that are specified velopmental switch genes, but they cannot be consid- during their development. Drosophila melanogaster ered specifically as behavioral switch genes. A switch males (but not females) perform an elaborate and in- gene for a sexual behavior should act to specify either nate courtship ritual directed toward females (but not male or female behavior, irrespective of the overall sex- males). Male courtship requires products of the fruit- ual phenotype of the animal. A candidate for such a less (fru) gene, which is spliced differently in males gene is the fruitless (fru) gene of Drosophila, which is and females. We have generated alleles of fru that are intimately linked to male sexual orientation and beha- constitutively spliced in either the male or the female vior (Baker et al., 2001).
    [Show full text]
  • Male-Specific Fruitless Isoforms Have Different Regulatory Roles Conferred
    Dalton et al. BMC Genomics 2013, 14:659 http://www.biomedcentral.com/1471-2164/14/659 RESEARCH ARTICLE Open Access Male-specific Fruitless isoforms have different regulatory roles conferred by distinct zinc finger DNA binding domains Justin E Dalton1†, Justin M Fear2,3,4†, Simon Knott5, Bruce S Baker6, Lauren M McIntyre3,4 and Michelle N Arbeitman1,7* Abstract Background: Drosophila melanogaster adult males perform an elaborate courtship ritual to entice females to mate. fruitless (fru), a gene that is one of the key regulators of male courtship behavior, encodes multiple male-specific isoforms (FruM). These isoforms vary in their carboxy-terminal zinc finger domains, which are predicted to facilitate DNA binding. Results: By over-expressing individual FruM isoforms in fru-expressing neurons in either males or females and assaying the global transcriptional response by RNA-sequencing, we show that three FruM isoforms have different regulatory activities that depend on the sex of the fly. We identified several sets of genes regulated downstream of FruM isoforms, including many annotated with neuronal functions. By determining the binding sites of individual FruM isoforms using SELEX we demonstrate that the distinct zinc finger domain of each FruM isoforms confers different DNA binding specificities. A genome-wide search for these binding site sequences finds that the gene sets identified as induced by over-expression of FruM isoforms in males are enriched for genes that contain the binding sites. An analysis of the chromosomal distribution of genes downstream of FruM shows that those that are induced and repressed in males are highly enriched and depleted on the X chromosome, respectively.
    [Show full text]
  • Functional Analysis of Fruitless Gene Expression by Transgenic Manipulations of Drosophila Courtship
    Functional analysis of fruitless gene expression by transgenic manipulations of Drosophila courtship Adriana Villella*, Sarah L. Ferri, Jonathan D. Krystal, and Jeffrey C. Hall* Department of Biology, Brandeis University, MS-008, Waltham, MA 02454 This contribution is part of the special series of Inaugural Articles by members of the National Academy of Sciences elected on April 29, 2003. Contributed by Jeffrey C. Hall, August 15, 2005 A gal4-containing enhancer–trap called C309 was previously revealed that C309 drives marker expression in a widespread shown to cause subnormal courtship of Drosophila males toward manner (18). Therefore, we sought to correlate various CNS females and courtship among males when driving a conditional regions in which this transgene is expressed with its effects on disrupter of synaptic transmission (shiTS). We extended these male behavior, emphasizing a search for ‘‘C309 neurons’’ that manipulations to analyze all features of male-specific behavior, might overlap with elements of the FRUM pattern. including courtship song, which was almost eliminated by driving We also entertained the possibility that the C309͞shiTS com- shiTS at high temperature. In the context of singing defects and bination causes a mere caricature of fruitless-like behavior. homosexual courtship affected by mutations in the fru gene, a Therefore, what would be the courtship effects of C309 driving tra-regulated component of the sex-determination hierarchy, we a transgene that produces the female form of the transformer found a C309͞traF combination also to induce high levels of gene product? This TRA protein participates in posttranscrip- courtship between pairs of males and ‘‘chaining’’ behavior in tional control of fru’s primary ‘‘sex transcript,’’ so that FRUM groups; however, these doubly transgenic males sang normally.
    [Show full text]
  • Fruitless Mutant Male Mosquitoes Gain Attraction to Human Odor
    RESEARCH ARTICLE Fruitless mutant male mosquitoes gain attraction to human odor Nipun S Basrur1*, Maria Elena De Obaldia1, Takeshi Morita1, Margaret Herre1,2, Ricarda K von Heynitz1†, Yael N Tsitohay1, Leslie B Vosshall1,2,3* 1Laboratory of Neurogenetics and Behavior, The Rockefeller University, New York, United States; 2Kavli Neural Systems Institute, New York, United States; 3Howard Hughes Medical Institute, New York, United States Abstract The Aedes aegypti mosquito shows extreme sexual dimorphism in feeding. Only females are attracted to and obtain a blood-meal from humans, which they use to stimulate egg production. The fruitless gene is sex-specifically spliced and encodes a BTB zinc-finger transcription factor proposed to be a master regulator of male courtship and mating behavior across insects. We generated fruitless mutant mosquitoes and showed that males failed to mate, confirming the ancestral function of this gene in male sexual behavior. Remarkably, fruitless males also gain strong attraction to a live human host, a behavior that wild-type males never display, suggesting that male mosquitoes possess the central or peripheral neural circuits required to host-seek and that removing fruitless reveals this latent behavior in males. Our results highlight an unexpected repurposing of a master regulator of male-specific sexual behavior to control one module of female-specific blood-feeding behavior in a deadly vector of infectious diseases. *For correspondence: [email protected] (NSB); [email protected] (LBV) Introduction Across animals, males and females of the same species show striking differences in behavior. Male Present address: † Technical Paradisaeidae birds-of-paradise perform an elaborate courtship dance to seduce prospective female University of Munich, TUM partners, contorting their bodies in forms resembling flowers, ballerinas, and smiling faces School of Medicine, Munich, Germany (Scholes, 2008).
    [Show full text]