In summary, this novel set of tools allows to characterize mRNA distribution in astrocytes and microglia in physiological or pathological settings. © 2020. Published by The Company of Biologists Ltd.All cells establish and maintain an axis of polarity that is critical for cell shape and progression through the cell cycle. A well-studied example of polarity establishment is bud emergence in yeast, which is controlled by the Rho GTPase Cdc42p. The prevailing view of bud emergence does not account for regulation by extrinsic cues. Here, we show that the filamentous growth MAPK pathway (fMAPK) regulates bud emergence under nutrient-limiting conditions. The fMAPK pathway regulated the expression of polarity targets including a direct effector of Cdc42p, Gic2p. The fMAPK pathway also stimulated GTP-Cdc42p levels, which is a critical determinant of polarity establishment. The fMAPK pathway activity was spatially restricted to bud sites and active during the period of the cell cycle leading up to bud emergence. Time-lapse fluorescence microscopy showed that the fMAPK pathway stimulated the rate of bud emergence during filamentous growth. Unregulated activation of the fMAPK pathway induced multiple rounds of symmetry breaking inside the growing bud. Collectively, our findings identify a new regulatory aspect of bud emergence that sensitizes this essential cellular process to external cues. © 2020. Published by The Company of Biologists Ltd.Germ cells use both positive and negative mRNA translational control to regulate gene expression that drives their differentiation into gametes. mRNA translational control is mediated by RNA-binding proteins, miRNAs, and translation initiation factors. We have uncovered the discrete roles of two translation initiation factor eIF4E isoforms (IFE-1 and -3) that bind m7GTP mRNA caps during C. elegans germline development. IFE-3 plays important roles in germline sex determination (GSD), where it promotes oocyte cell fate and is dispensable for spermatogenesis. IFE-3 is expressed throughout the germline, localizes to germ granules but distinct from IFE-1 and PGL-1, and facilitates oocyte growth and viability. This contrasts with the robust expression of IFE-1 in spermatocytes, the isoform that resides within P granules in spermatocytes and oocytes, and promotes late spermatogenesis. Each eIF4E is localized by its cognate eIF4E binding protein (IFE-1PGL-1 and IFE-3IFET-1). IFE-3 and IFET-1 regulate translation of several GSD mRNAs, but not those under control of IFE-1. Distinct mutant phenotypes, in vivo localization, and differential mRNA translation suggest independent dormant and active periods for each eIF4E isoform in the germline.  https://www.selleckchem.com/products/iruplinalkib.html  © 2020. Published by The Company of Biologists Ltd.Intracellular survival of Leishmania donovani demands rapid production of host ATP for its sustenance. However, gradual decrease in intracellular ATP in spite of increased glycolysis suggested ATP efflux during infection. Accordingly, extracellular ATP showed an increase and pannexin-1 was found to be the major ATP exporter in infected condition. Extracellular ATP also showed a gradual decrease after initial increase and analysing cell surface ATP-degrading enzymes revealed induction of the ectonucleotidases, CD39 and CD73. Ectonucleotidase mediated ATP degradation led to increased extracellular adenosine (eADO) and inhibiting CD39 and CD73 in infected cells decreased adenosine concentration and parasite survival, documenting importance of adenosine in infection. Inhibiting adenosine uptake by cells did not affect parasite survival implicating eADO might exert its effect through receptor-mediated signaling. Leishmania indeed induced expression of adenosine receptors A2AR and A2BR, both being important for anti-inflammatory responses. Treating infected BALB/c mice with CD39 and CD73 inhibitors resulted in decreased parasite burden and increased host-favourable cytokine production. Collectively, these observations indicate that infection-induced ATP is exported and after converting into adenosine exerts receptor-mediated signaling for propagating infection. © 2020. Published by The Company of Biologists Ltd.F-actin dynamics are known to control insulin secretion but the point of intersection with the stimulus-secretion cascade is unknown. Here, using multiphoton imaging of β cells isolated from Lifeact-GFP transgenic mice, we show glucose stimulation does not cause global changes in subcortical F-actin. Instead, we observe spatially discrete and transient F-actin changes around each fusing granule. This F-actin remodelling is dependent on actin nucleation and is observed for granule fusion induced by either glucose or high potassium stimulation. Using GFP-labelled proteins we identify local enrichment of Arp3, dynamin and clathrin, all occurring after granule fusion, suggesting early recruitment of an endocytic complex to the fusing granules. Block of Arp2/3 activity with drugs or shRNA inhibit F-actin coating, traps granules at the cell membrane and reduces insulin secretion. Block of formin-mediated actin nucleation also blocks F-actin coating but has no effect on insulin secretion. We conclude that local Arp2/3 dependent actin nucleation at the sites of granule fusion plays an important role in post-fusion granule dynamics and in the regulation of insulin secretion. © 2020. Published by The Company of Biologists Ltd.The mitochondrial DNA of Trypanosoma brucei and related parasites is a catenated network containing thousands of minicircles and tens of maxicircles called kinetoplast DNA (kDNA). Replication of the single nucleoid requires at least three DNA polymerases (POLIB, POLIC, and POLID) each having discrete localization near the kDNA during S phase. POLIB and POLID have roles in minicircle replication while the specific role of POLIC in kDNA maintenance is less clear. Here, we use an RNAi-complementation system to dissect the functions of the distinct POLIC domains the conserved family A DNA polymerase domain (POLA) and the uncharacterized N-terminal region (UCR). While RNAi complementation with wild-type POLIC restored kDNA content and cell cycle localization, active site point mutations in the POLA domain impaired minicircle replication similarly to POLIB and POLID depletions. Complementation with POLA domain alone abolished POLIC foci formation and partially rescued the RNAi phenotype. Furthermore, we provide evidence of a crucial role for the UCR in cell cycle localization that facilitates proper distribution of progeny networks.