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Hydrocephalus is a prevalent condition among newborns leading to substantial neurocognitive and motor impairment. Novel therapies are needed to supplant invasive surgeries, but identifying targetable cells and pathways remains a hurdle to devising alternative pharmacological options. Multiciliated ependymal cells (MECs) promote cerebrospinal fluid flow within brain ventricles, and their dysfunction is associated with various forms of hydrocephalus. Here we show that an acute exposure to TNF-α strongly impairs the conversion of ependymal cell radial glial progenitors (ecRGPs) into MECs. Inhibition of MEC differentiation was correlated with elevated expression levels of notch pathway effectors normally downregulated prior to the transition of ecRGPs into MECs. TNF-α inhibitedMulticilingene upregulation along with downstream genes critical for centriole amplification and multicilia formation, resulting in cells with greatly diminished basal bodies and multicilia. Treatment with notch inhibitor DBZ, either in parallel with TNF-α or sequentially days later, rescued MEC differentiation and expression of genes required for multicilia formation. These results provide a rationale for how TNFα can impair MEC development, and they offer a targetable pathway to the treatment of some forms of hydrocephalus.

SUMMARYA role for macronutrient metabolism in learning and memory is supported by numerous epidemiological studies. The Ppm1k gene encodes the branched-chain keto acid dehydrogenase (BCKDH) phosphatase that promotes the metabolism of branched-chain amino acids (BCAA). Here we show that nervous system deletion of Ppm1k in mice increases BCAA levels in brain tissue but not in plasma. These mice have significant impairments in working memory accompanied by a robust accumulation of DCX+/NeuroD1+ immature neurons within the dentate gyrus granule cell layer. Through single cell RNA sequencing and pathway analysis we identified substantial increases in transit-amplifying cells and immature neurons along with activated hedgehog signaling in Ppm1k deficient primary neural stem cells (NSCs). Inhibition of mTOR signaling reversed the effects of Ppm1k deletion on neuronal progenitor gene activation in primary NSCs. Together our findings uncover a new molecular link between BCAA metabolism, hippocampal neurogenesis, and cognitive performance.

Sclerodermus brevicornis is a parasitoid that exhibits cooperative multi-foundress brood production. Prior work showed that the time lag to paralysis of small-sized hosts is shorter when co-foundress relatedness is higher and predicted that the greater risks and greater benefits of attacking larger hosts would combine with co-foundress relatedness to determine the limits to the size of a host that a female is selected to attack as a public good. It was also predicted that the time to host attack would be affected by an interaction between host size and relatedness. Here, we show empirically that both host size and kinship affect S. brevicornis reproduction and that they interact to influence the timing of host attack. We also find effects of co-foundress relatedness after hosts have been suppressed successfully. A public goods model using parameters estimated for S. brevicornis again suggests that selection for individual foundresses to attack and, if successful, to share hosts will be dependent on both the size of the host and the relatedness of the foundresses to any co-foundresses present. Females will not be selected to bear the individual cost of a public good when hosts are large and dangerous or when their relatedness to the co-foundress is low. We conclude that although reproductive behaviours exhibited by Sclerodermus females can be cooperative, they are unlikely to be exhibited without reference to kinship or to the risks involved in attempting to suppress and share large and dangerous hosts.

Explanations for the highest levels of sociality typically invoke the concept of inclusive fitness. Sclerodermus, a genus of parasitoid hymenopterans, is quasi-social, exhibiting cooperative brood care without generational overlap or apparent division of labour. Foundress females successfully co-exploit hosts that are too large to suppress when acting alone and the direct fitness benefits of collective action may explain their cooperation, irrespective of kinship. However, cooperation in animal societies is seldom free of conflicts of interest between social partners, especially when their relatedness, and thus their degree of shared evolutionary interests, is low. We screened components of the life-history of Sclerodermus brevicornis for effects of varying co-foundress number and relatedness on cooperative reproduction. We found that the time taken to paralyse standard-sized hosts is shorter when co-foundress number and/or relatedness is higher. This suggests that, while females must access a paralysed host in order to reproduce, individuals are reluctant to take the risk of host attack unless the benefits will be shared with their kin. We used Hamilton’s rule and prior data from studies that experimentally varied the sizes of hosts presented to congeners to explore how the greater risks and greater benefits of attacking larger hosts could combine with relatedness to determine the sizes of hosts that individuals are selected to attack as a public good. From this, we predict that host size and relatedness will interact to affect the timing of host paralysis; we test this prediction in the accompanying study.

ispartof: J Neonatal Perinatal Med vol:15 issue:2 pages:411-426 ispartof: location:Netherlands status: published

SUMMARY Specialized microenvironments, called niches, control adult stem cell proliferation and differentiation. The brain lateral ventricular (LV) neurogenic niche is generated from distinct postnatal radial glial progenitors (pRGPs), giving rise to adult neural stem cells (NSCs) and niche ependymal cells (ECs). Cellular-intrinsic programs govern stem versus supporting cell maturation during adult niche assembly, but how they are differentially initiated within a similar microenvironment remains unknown. Using chemical approaches, we discovered that EGFR signaling powerfully inhibits EC differentiation by suppressing multiciliogenesis. We found that EC pRGPs actively terminated EGF activation through receptor redistribution away from CSF-contacting apical domains and that randomized EGFR membrane targeting blocked EC differentiation. Mechanistically, we uncovered spatiotemporal interactions between EGFR and endocytic adaptor protein Numb. Ca2+-dependent basolateral targeting of Numb is necessary and sufficient for proper EGFR redistribution. These results reveal a previously unknown cellular mechanism for neighboring progenitors to differentially engage environmental signals, initiating adult stem cell niche assembly., Graphical Abstract, In Brief Constructing adult stem niches from postnatal progenitors requires differential cellular interpretations of similar microenvironmental signals. Abdi et al. show that in an EGF-rich environment needed for adult neural stem cell proliferation, a subpopulation of postnatal progenitors downregulates EGFR signaling via targeted receptor trafficking, promoting multiciliated ependymal niche cell differentiation.

Kinship among interacting individuals is often associated with sociality and also with sex ratio effects. Parasitoids in the bethylid genus Goniozus are sub-social, with single foundress females exhibiting post-ovipositional maternal care via short-term aggressive host and brood defence against conspecific females. Due to local mate competition (LMC) and broods normally being produced by a single foundress, sex ratios are female-biased. Contests between adult females are, however, not normally fatal, and aggression is reduced when competing females are kin, raising the possibility of multi-foundress reproduction on some hosts. Here, we screen for further life-history effects of kinship by varying the numbers and relatedness of foundresses confined together with a host resource and also by varying the size of host. We confined groups of 1-8 Goniozus nephantidis females together with a host for 5+ days. Multi-foundress groups were either all siblings or all nonsiblings. Our chief expectations included that competition for resources would be more intense among larger foundress groups but diminished by both larger host size and closer foundress relatedness, affecting both foundress mortality and reproductive output. From classical LMC theory, we expected that offspring group sex ratios would be less female-biased when there were more foundresses, and from extended LMC theory, we expected that sex ratios would be more female-biased when foundresses were close kin. We found that confinement led to the death of some females (11% overall) but only when host resources were most limiting. Mortality of foundresses was less common when foundresses were siblings. Developmental mortality among offspring was considerably higher in multi-foundress clutches but was unaffected by foundress relatedness. Groups of sibling foundresses collectively produced similar numbers of offspring to nonsibling groups. There was little advantage for individual females to reproduce in multi-foundress groups: single foundresses suppressed even the largest hosts presented and had the highest per capita production of adult offspring. Despite single foundress reproduction being the norm, G. nephantidis females in multi-foundress groups appear to attune sex allocation according to both foundress number and foundress relatedness: broods produced by sibling foundresses had sex ratios similar to broods produced by single foundresses (ca. 11% males), whereas the sex ratios of broods produced by nonsibling females were approximately 20% higher and broadly increased with foundress number. We conclude that relatedness and host size may combine to reduce selection against communal reproduction on hosts and that, unlike other studied parasitoids, G. nephantidis sex ratios conform to predictions of both classical and extended LMC theories.

Several recent reports have found a connection between specific aminoacyl-tRNA synthetases and the regulation of angiogenesis. As this new area of research is explored, it is important to have reliable assays to assess the specific angiogenesis functions of these enzymes. This review provides information about specific in vitro and in vivo methods that were used to assess the angiogenic functions of threonyl-tRNA synthetase including endothelial cell migration and tube assays as well as chorioallantoic membrane and tumor vascularization assays. The theory and discussion include best methods of analysis and quantification along with the advantages and limitations of each type of assay.

SUMMARY Dendrites often exhibit structural changes in response to local inputs. Although mechanisms that pattern and maintain dendritic arbors are becoming clearer, processes regulating regrowth, during context-dependent plasticity or after injury, remain poorly understood. We found that a class of Drosophila sensory neurons, through complete pruning and regeneration, can elaborate two distinct dendritic trees, innervating independent sensory fields. An expression screen identified Cysteome proteinase-1 (Cp1) as a critical regulator of this process. Unlike known ecdysone effectors, Cp1-mutant ddaC neurons pruned larval dendrites normally but failed to regrow adult dendrites. Cp1 expression was upregulated/concentrated in the nucleus during metamorphosis, controlling production of a truncated Cut homeodomain transcription factor. This truncated Cut, but not the full-length protein, allowed Cp1-mutant ddaC neurons to regenerate higher-order adult dendrites. These results identify a molecular pathway needed for dendrite regrowth after pruning, which allows the same neuron to innervate distinct sensory fields.

During mammalian brain development, radial glial progenitors balance between proliferation and differentiation to generate the laminated cortical layers in a temporally precise fashion. Defects in the individual steps going into this complex organogenesis can result in cortical malformations and human nervous system disorders. In this issue of Genes & Development, Liu and colleagues (pp. 763–780) present experimental evidence that an evolutionarily conserved cellular polarity gene, Pard3 (partitioning-defective 3), controls the balance of radial glial proliferation and differentiation through interaction with the Hippo signal transduction pathway. Conditional deletion of Pard3 in the developing rodent cortex resulted in striking subcortical band heterotopia, reminiscent of a severe form of human cortical malformation.

Background: E-cadherin targets to the epithelial lateral membrane. Results: Ankyrin-G and clathrin cooperate to control E-cadherin localization. Conclusion: A multifunction motif on E-cadherin interacts with multiple pathways. Significance: This finding provides novel insights into how basolateral targeting motifs function., We report a highly conserved motif in the E-cadherin juxtamembrane domain that determines apical-lateral polarity by conferring both restricted mobility at the lateral membrane and transcytosis of apically mis-sorted protein to the lateral membrane. Mutations causing either increased lateral membrane mobility or loss of apical-lateral transcytosis result in partial mis-sorting of E-cadherin in Madin-Darby canine kidney cells. However, loss of both activities results in complete loss of polarity. We present evidence that residues required for restricted mobility mediate retention at the lateral membrane through interaction with ankyrin-G, whereas dileucine residues conferring apical-lateral transcytosis act through a clathrin-dependent process and function in an editing pathway. Ankyrin-G interaction with E-cadherin is abolished by the same mutations resulting in increased E-cadherin mobility. Clathrin heavy chain knockdown and dileucine mutation of E-cadherin both cause the same partial loss of polarity of E-cadherin. Moreover, clathrin knockdown causes no further change in polarity of E-cadherin with dileucine mutation but does completely randomize E-cadherin mutants lacking ankyrin-binding. Dileucine mutation, but not loss of ankyrin binding, prevented transcytosis of apically mis-sorted E-cadherin to the lateral membrane. Finally, neurofascin, which binds ankyrin but lacks dileucine residues, exhibited partial apical-lateral polarity that was abolished by mutation of its ankyrin-binding site but was not affected by clathrin knockdown. The polarity motif thus integrates complementary activities of lateral membrane retention through ankyrin-G and apical-lateral transcytosis of mis-localized protein through clathrin. Together, the combination of retention and editing function to ensure a high fidelity steady state localization of E-cadherin at the lateral membrane.

Biological systems are constantly under mechanical stress either during movement or when acted upon by external forces. The identification of proteins motifs that behave as biological springs will be important for understanding how cells respond to mechanical stimuli and can also propel the design of non-biological nanomaterials. We report here identification of a large class of alpha-helical spiral or solenoid-shaped proteins comprised of ANK-R, ARM, or HEAT repeats that rapidly and forcefully refold following stretching. Each of these repeats unfolds and refolds in equilibrium through discrete events involving individual repeats or their alpha-helical subunits. We also present evidence for the capture of unfolding/refolding transient events while stretching or relaxing by AFM and analyze differences in refolding lengths and forces for each repeat. This class of stacked helical-repeats behave as molecular nanosprings, are likely important for cellular mechanosensation, and can be used as platforms for structural elements of nanomechanical systems based on proteins. Supported by the NIH (PEM) and HHMI (VB).

SummaryThe rodent subventricular/subependymal zone (SVZ/SEZ) houses neural stem cells (NSCs) that generate olfactory bulb interneurons. It is unclear how the SVZ environment sustains neuronal production into adulthood. We discovered that the adapter molecule Ankyrin-3 (Ank3) is specifically upregulated in ventricular progenitors destined to become ependymal cells, but not in NSCs, and is required for SVZ niche assembly through progenitor lateral adhesion. Furthermore, we found that Ank3 expression is controlled by Foxj1, a transcriptional regulator of multicilia formation, and genetic deletion of this pathway led to complete loss of SVZ niche structure. Interestingly, radial glia continued to transition into postnatal NSCs without this niche. However, inducible deletion of Foxj1-Ank3 from mature SVZ ependyma resulted in dramatic depletion of neurogenesis. Targeting a pathway regulating ependymal organization/assembly and showing its requirement for new neuron production, our results have important implications for environmental control of adult neurogenesis and harvesting NSCs for replacement therapy.

Palmitoylation of ankyrin-G and interaction of βII-spectrin with phosphoinositides are necessary for ankyrin-G–βII-spectrin localization in membrane subdomains during lateral membrane assembly in columnar epithelial cells., Ankyrin-G and βII-spectrin colocalize at sites of cell–cell contact in columnar epithelial cells and promote lateral membrane assembly. This study identifies two critical inputs from lipids that together provide a rationale for how ankyrin-G and βII-spectrin selectively localize to Madin-Darby canine kidney (MDCK) cell lateral membranes. We identify aspartate-histidine-histidine-cysteine 5/8 (DHHC5/8) as ankyrin-G palmitoyltransferases required for ankyrin-G lateral membrane localization and for assembly of lateral membranes. We also find that βII-spectrin functions as a coincidence detector that requires recognition of both ankyrin-G and phosphoinositide lipids for its lateral membrane localization. DHHC5/8 and βII-spectrin colocalize with ankyrin-G in micrometer-scale subdomains within the lateral membrane that are likely sites for palmitoylation of ankyrin-G. Loss of either DHHC5/8 or ankyrin-G–βII-spectrin interaction or βII-spectrin–phosphoinositide recognition through its pleckstrin homology domain all result in failure to build the lateral membrane. In summary, we identify a functional network connecting palmitoyltransferases DHHC5/8 with ankyrin-G, ankyrin-G with βII-spectrin, and βII-spectrin with phosphoinositides that is required for the columnar morphology of MDCK epithelial cells.

Postnatal/adult neural stem cells (NSCs) within the rodent subventricular zone (SVZ; also called subependymal zone) generate doublecortin (Dcx)(+) neuroblasts that migrate and integrate into olfactory bulb circuitry. Continuous production of neuroblasts is controlled by the SVZ microenvironmental niche. It is generally thought that enhancing the neurogenic activities of endogenous NSCs may provide needed therapeutic options for disease states and after brain injury. However, SVZ NSCs can also differentiate into astrocytes. It remains unclear whether there are conditions that favour astrogenesis over neurogenesis in the SVZ niche, and whether astrocytes produced there have different properties compared with astrocytes produced elsewhere in the brain. Here we show in mice that SVZ-generated astrocytes express high levels of thrombospondin 4 (Thbs4), a secreted homopentameric glycoprotein, in contrast to cortical astrocytes, which express low levels of Thbs4. We found that localized photothrombotic/ischaemic cortical injury initiates a marked increase in Thbs4(hi) astrocyte production from the postnatal SVZ niche. Tamoxifen-inducible nestin-creER(tm)4 lineage tracing demonstrated that it is these SVZ-generated Thbs4(hi) astrocytes, and not Dcx(+) neuroblasts, that home-in on the injured cortex. This robust post-injury astrogenic response required SVZ Notch activation modulated by Thbs4 via direct Notch1 receptor binding and endocytosis to activate downstream signals, including increased Nfia transcription factor expression important for glia production. Consequently, Thbs4 homozygous knockout mice (Thbs4(KO/KO)) showed severe defects in cortical-injury-induced SVZ astrogenesis, instead producing cells expressing Dcx migrating from SVZ to the injury sites. These alterations in cellular responses resulted in abnormal glial scar formation after injury, and significantly increased microvascular haemorrhage into the brain parenchyma of Thbs4(KO/KO) mice. Taken together, these findings have important implications for post-injury applications of endogenous and transplanted NSCs in the therapeutic setting, as well as disease states where Thbs family members have important roles.