Project description:We aimed at identifying the genes regulated by wounding in Anopheles gambiae. Gene expression was compared between wounded and non-wounded mosquitoes, 3h after wounding. Wounding was induced by the injection of dsLacZ using a thin glass needle.
Project description:Unilateral injections of 6-hydroxydopamine into the medial forebrain bundle (MFB) is used extensively as a model of Parkinson’s disease. The present experiments examined whether a single injection of methamphetamine (METH) (2.5 mg/kg) could still influence striatal gene expression after 6-hydroxydopamine (DA)-induced destruction of the nigrostriatal dopaminergic pathway in the rat. Unilateral injections of 6-hydroxydopamine into the MFB resulted in total striatal dopamine depletion on the lesioned side. This injection also caused decreased striatal serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) levels. DA depletion was associated with increases in 5-HIAA/5-HT ratios which were potentiated by the METH injection. Microarray analyses revealed changes (+ 1.7-fold, p < 0.025) in the expression of 67 genes on the lesioned side in comparison to the intact side of the saline-treated hemiparkinsonian animals. These include follistatin, neuromedin U, and tachykinin 2 which were up-regulated. METH administration caused increases in the expression of several genes including c-fos, Egr1, and NOR-1 on the intact side. On the DA-depleted side, METH administration also increased the expression of 61 genes including Pdgfd and COX-2. There were METH-induced changes in 16 genes that were common in the DA-innervated and DA-depleted sides. These include c-fos and NOR-1 which show greater changes on the normal DA side. The present study documents METH-mediated DA-independent transcriptional alterations of several genes in the DA-denervated striatum. Our results also implicate serotonin as an important player in METH-induced gene expression because the METH injection also caused significant increases in 5-HIAA/5-HT ratios on the DA-depleted side. 31 samples. MNL (6), ML (6), SNL (5), SL (6), CS (4), CM (4)
Project description:Toll pathway is a key mediator of antiplasmodial immunity and its mechanism of action is dependent on hemocytes (mosquito white blood cells). Anopheles gambiae were injected with dsRNA for Cactus, an inhibitor of the Toll pathway. Cactus silencing over activates the Toll pathway. The goal of this experiment is to evaluate hemocyte transcriptional changes between control mosquitoes injected with a non-related dsRNA (dsLacZ and dsCactus injected mosquitoes. We also divide hemocytes from control and experimental conditions into 2 groups: bound (hemocytes that attach to glass - granulocyte populations) and unbound (hemocytes that remain in suspension - prohemocytes and oenocytoids). Files from Unbound fractions are labelled: UB and from bound fractions are labelled B. Some of the samples were run twice and have 2 gz files.
Project description:Protein phosphorylation plays a critical role during the development of malaria parasites. Here, we performed a functional analysis of the Plasmodium berghei Ser/Thr protein phosphatase 6 (PbPP6), which was associated with the plasma membrane of macrogametes and ookinetes. Compared to wild-type P. berghei, genetic disruption/deletion of the pbpp6 gene (∆pbpp6) reduced the asexual growth of the parasites and prolonged the survival of ∆pbpp6-infected mice. The ∆pbpp6 parasites showed impaired gametogenesis, particularly affecting male gametogenesis, which substantially decreased ookinete formation and transmission to mosquitoes. RNA-seq assays revealed that pbpp6 disruption led to over 11-fold down-regulation of the nek3 gene, a regulator of MAPK2 in the PKG-Ca2+ signaling cascade during male gametogenesis. Several PDEs (α, γ, δ) were also affected, indicating that PbPP6 plays a role in the cGMP-PKG-Ca2+ signaling pathway. Additionally, we observed altered expression of messenger ribonucleoproteins in the Δpbpp6 parasites, which may affect translational repression of stored mRNAs in female gametocytes and post-fertilization development in mosquitoes. Consistent with the dysregulated GO terms related to the cGMP-PKG-Ca2+ signaling cascade observed in RNA-seq analysis, the activated gametocytes of ∆pbpp6 exhibited significantly reduced levels of intracellular cGMP, cytosolic Ca2+ mobilization, and DNA replication. Phosphoproteomic analysis detected increased phosphorylation at the Ser508 site of guanylyl cyclase alpha (GCα), suggesting that PbPP6 regulates cGMP-PKG-Ca2+ signaling cascades by modulating the activity of GCα during gametogenesis. This study highlights the potential of targeting PP6 to disrupt malaria transmission.
Project description:Protein phosphorylation plays a critical role during the development of malaria parasites. Here, we performed a functional analysis of the Plasmodium berghei Ser/Thr protein phosphatase 6 (PbPP6), which was associated with the plasma membrane of macrogametes and ookinetes. Compared to wild-type P. berghei, genetic disruption/deletion of the pbpp6 gene (∆pbpp6) reduced the asexual growth of the parasites and prolonged the survival of ∆pbpp6-infected mice. The ∆pbpp6 parasites showed impaired gametogenesis, particularly affecting male gametogenesis, which substantially decreased ookinete formation and transmission to mosquitoes. RNA-seq assays revealed that pbpp6 disruption led to over 11-fold down-regulation of the nek3 gene, a regulator of MAPK2 in the PKG-Ca2+ signaling cascade during male gametogenesis. Several PDEs (α, γ, δ) were also affected, indicating that PbPP6 plays a role in the cGMP-PKG-Ca2+ signaling pathway. Additionally, we observed altered expression of messenger ribonucleoproteins in the Δpbpp6 parasites, which may affect translational repression of stored mRNAs in female gametocytes and post-fertilization development in mosquitoes. Consistent with the dysregulated GO terms related to the cGMP-PKG-Ca2+ signaling cascade observed in RNA-seq analysis, the activated gametocytes of ∆pbpp6 exhibited significantly reduced levels of intracellular cGMP, cytosolic Ca2+ mobilization, and DNA replication. Phosphoproteomic analysis detected increased phosphorylation at the Ser508 site of guanylyl cyclase alpha (GCα), suggesting that PbPP6 regulates cGMP-PKG-Ca2+ signaling cascades by modulating the activity of GCα during gametogenesis. This study highlights the potential of targeting PP6 to disrupt malaria transmission.