Jian Wang

How neural stem cells regulate their proliferative potential and lineage diversity is a central problem in developmental neurobiology. Drosophila Mushroom bodies (MBs), centers of olfactory learning and memory, are generated by a specific set of neuroblasts (Nbs) that are born in the embryonic stage and continuously proliferate till the end of the pupal stage. Although MB presents an excellent model for studying neural stem cell proliferation, the genetic and molecular mechanisms that control the unique proliferative characteristics of the MB Nbs are largely unknown. Further, the signaling cues controlling cell cycle regulators to promote cell cycle progression in MB Nbs remain poorly understood. Here, we report that JAK/STAT signaling pathway is required for the proliferation activity and maintenance of MB Nbs. Loss of JAK/STAT activity severely reduces the later-born MB neuron types and leads to premature neuroblast termination, which can be rescued by tissue-specific overexpression of CycE and diap1 . Higher JAK/STAT pathway activity in MB results in more neurons, without producing supernumerary Nbs. Furthermore, we show that JAK/STAT signaling effector Stat92E directly regulates CycE transcription in MB Nbs. Finally, MB Nb clones of loss or excess CycE phenocopy those of decreased or increased JAK/STAT signaling pathway activities. We conclude that JAK/STAT signaling controls MB Nb proliferative activity through directly regulating CycE expression to control cell cycle progression.

Odorant-binding proteins (OBPs) are important in insect chemical communication. The objective of this research was to identify the functions of two OBPs in Sitophilus zeamais. qRT-PCR and western blot (WB) were performed to investigate the expression profiles at the transcript and protein levels, respectively. Fluorescence competitive binding assays were used to measure the ability of the OBPs to bind to host volatiles, and a Y-tube olfactometer was used to verify the results (attraction/no response) via behavioral experiments. The RNAi was used to verify the function by knocking down the ability of proteins to bind odorants. qRT-PCR showed the highest expression SzeaOBP1 and SzeaOBP28 at the low-instar larva (LL) and eclosion adult (EA) stages, respectively. WB showed that both SzeaOBP1 and SzeaOBP28 were highly expressed in the EA stage. Fluorescence competitive binding assays indicated that SzeaOBP1 exhibited extremely high binding affinity with cetanol. SzeaOBP28 exhibited a pronounced binding affinity for 4-hydroxy-3-methoxybenzaldehyde. The behavioral experiment showed that the adult S. zeamais responded strongly to 4-hydroxy-3-methoxybenzaldehyde and valeraldehyde from Sorghum bicolor. The RNAi knockdown individuals displayed behavioral differences between normal insects and dsRNA (SzeaOBP1)-treated insects. We infer that they both have functions in perception and recognition of host volatiles, whereas SzeaOBP28 may also have other functions.

During Bt transgenic rice storage, Bt Cry1Ab/Cry1Ac fused protein is exposed to the maize weevil Sitophilus zeamais and the parasitoid wasp Theocolax elegans. We have carried out a long-term risk assessment for Bt rice to these non-target organisms in the storehouse. Effects of Bt rice on S. zeamais and T. elegans have been carefully detected in a laboratory experiment of over 5 years. The survival, development, fecundity, and longevity of the maize weevil were compared between Bt rice and non-Bt rice treatments for every 5 generations from generation 1 to 25. Moreover, the development, adult body size and sex ratio of T. elegans were compared between them parasitizing S. zeamais feeding on Bt rice or non-Bt rice. We found that although Bt Cry1Ab/Cry1Ac fused protein exists in the Bt rice grains and S. zeamais digestive tracts, Bt rice is not harmful to the maize weevil S. zeamais and its parasitoid T. elegans.

Our bioassays reviewed that antennae played crucial roles in the responses of maize weevil (Sitophilus zeamais) to food and sex volatiles. In order to identify the maize weevil odorant‐binding protein (OBP) genes, we analyzed its antennal transcriptome. In total, 21,587,928 high‐quality clean reads were obtained from RNA‐seq, 52,206 unigenes were assembled, and 25,744 unigenes showed significant similarity ( E value < 10 ⁻⁵) to known proteins in the NCBI nonredundant protein database. From those unigenes, we identified 41 candidate OBP proteins, which could be categorized into dimeric OBPs subfamily, minus‐C OBPs subfamily, and classical OBPs subfamily. Phylogenic analysis indicated that most maize weevil OBPs were closely related to their orthologues in other beetles of the Superfamily Curculionoidea. We further investigated the expression profiles of those candidate OBP genes by quantitative real‐time polymerase chain reaction. Twenty‐six of forty‐one maize weevil OBP genes were highly expressed in the antennae or other parts of the head. The rest were expressed in the legs, wings, or other tested tissues. The antennal transcriptomic data and candidate OBP genes described here provide a basis for the functional studies of the maize weevil chemical perception, which are potential novel targets for pest control strategies.

Corpus allatum (CA) ablation results in juvenile hormone (JH) deficiency and pupal lethality in Drosophila. The fly CA produces and releases three sesquiterpenoid hormones: JH III bisepoxide (JHB3), JH III, and methyl farnesoate (MF). In the whole body extracts, MF is the most abundant sesquiterpenoid, followed by JHB3 and JH III. Knockout of JH acid methyl transferase (jhamt) did not result in lethality; it decreased biosynthesis of JHB3, but MF biosynthesis was not affected. RNAi-mediated reduction of 3-hydroxy-3-methylglutaryl CoA reductase (hmgcr) expression in the CA decreased biosynthesis and titers of the three sesquiterpenoids, resulting in partial lethality. Reducing hmgcr expression in the CA of the jhamt mutant further decreased MF titer to a very low level, and caused complete lethality. JH III, JHB3, and MF function through Met and Gce, the two JH receptors, and induce expression of Kr-h1, a JH primary-response gene. As well, a portion of MF is converted to JHB3 in the hemolymph or peripheral tissues. Topical application of JHB3, JH III, or MF precluded lethality in JH-deficient animals, but not in the Met gce double mutant. Taken together, these experiments show that MF is produced by the larval CA and released into the hemolymph, from where it exerts its anti-metamorphic effects indirectly after conversion to JHB3, as well as acting as a hormone itself through the two JH receptors, Met and Gce. Methyl farnesoate (MF) is the immediate precursor of juvenile hormone (JH) III in the JH biosynthetic pathway, and lacks the epoxide moiety characteristic of JHs. The potential role of MF as a JH in arthropods has been an issue of a long-standing debate. In this report, comprehensive molecular genetics studies demonstrated that MF plays a dual role in regulating Drosophila metamorphosis. MF is produced by the larval CA and released into the hemolymph, from where it exerted its anti-metamorphic effects indirectly after conversion to JHB3, as well as acting as a hormone itself through a direct interaction with Met and Gce, the two JH receptors.

Drosophilapolyhomeotic (ph) is one of the important polycomb group genes that is linked to human cancer. In the mosaic eye imaginal discs, while ph^del, a null allele, causes only non-autonomous overgrowth, ph⁵⁰⁵, a hypomorphic allele, causes both autonomous and non-autonomous overgrowth. These allele-specific phenotypes stem from the different sensitivities of ph mutant cells to the Upd homologs that they secrete.

Juvenile hormone (JH) plays key roles in controlling insect growth and metamorphosis. However, relatively little is known about the JH signaling pathways. Until recent years, increasing evidence has suggested that JH modulates the action of 20-hydroxyecdysone (20E) by regulating expression of broad (br), a 20E early response gene, through Met/Gce and Kr-h1. To identify other genes involved in JH signaling, we designed a novel Drosophila genetic screen to isolate mutations that derepress JH-mediated br suppression at early larval stages. We found that mutations in three Wnt signaling negative regulators in Drosophila, Axin (Axn), supernumerary limbs (slmb), and naked cuticle (nkd), caused precocious br expression, which could not be blocked by exogenous JHA. A similar phenotype was observed when armadillo (arm), the mediator of Wnt signaling, was overexpressed. qRT-PCR revealed that Met, gce and Kr-h1expression was suppressed in the Axn, slmb and nkd mutants as well as in arm gain-of-function larvae. Furthermore, ectopic expression of gce restored Kr-h1 expression but not Met expression in the arm gain-of-function larvae. Taken together, we conclude that Wnt signaling cross-talks with JH signaling by suppressing transcription of Met and gce, genes that encode for putative JH receptors. The reduced JH activity further induces down-regulation of Kr-h1expression and eventually derepresses br expression in the Drosophila early larval stages.

Polycomb group (PcG) proteins are conserved epigenetic regulators that are linked to cancer in humans. However, little is known about how they control cell proliferation. Here, we report that mutant clones of the PcG gene polyhomeotic ( ph ) form unique single‐cell‐layer cavities that secrete three JAK/STAT pathway ligands, which in turn act redundantly to stimulate overproliferation of surrounding wild‐type cells. Notably, different ph alleles cause different phenotypes at the cellular level. Although the ph ‐null allele induces non‐autonomous overgrowth, an allele encoding truncated Ph induces both autonomous and non‐autonomous overgrowth. We propose that PcG misregulation promotes tumorigenesis through several cellular mechanisms.