The inbred mouse C57BL/6J is the reference strain for genome sequence and for most behavioral and physiological phenotypes. However, the International Knockout Mouse Consortium uses an embryonic stem cell line derived from a related C57BL/6N substrain. We found that C57BL/6N has a lower acute and sensitized response to cocaine and methamphetamine. We mapped a single causative locus and identified a nonsynonymous mutation of serine to phenylalanine (S968F) in Cytoplasmic FMRP interacting protein 2 (Cyfip2) as the causative variant. The S968F mutation destabilizes CYFIP2, and deletion of the C57BL/6N mutant allele leads to acute and sensitized cocaine-response phenotypes. We propose that CYFIP2 is a key regulator of cocaine response in mammals and present a framework to use mouse substrains to identify previously unknown genes and alleles regulating behavior.

https://science.sciencemag.org/content/sci/342/6165/1508.full.pdf

C57BL/6N Mutation in Cytoplasmic FMRP interacting protein 2 Regulates Cocaine Response

Local β-actin synthesis in growth cones of developing axons plays an important role in growth cone steering; however, the mRNA binding proteins required for this process are unknown. Here we used Zbp1 / Imp1 −/ − mice to test the hypothesis that zipcode binding protein 1 (ZBP1) is required for the regulation of β-actin mRNA transport and local translation underlying growth cone guidance. To address the biological function of ZBP1, we developed a novel in vitro turning assay with primary cortical neuron balls having axons >1 mm in length and demonstrate that growth cones of mammalian neurons exhibit protein synthesis-dependent attraction to either netrin-1 or brain-derived neurotrophic factor (BDNF). Interestingly, this attraction is lost in Zbp1 -deficient neurons. Furthermore, BDNF-stimulated β-actin mRNA localization was attenuated in Zbp1 -deficient neurons, which impaired enrichment of β-actin protein in the growth cone. Finally, using a photoconvertible translation reporter, we found that ZBP1 is necessary for netrin-1 stimulated local translation of β-actin mRNA in axonal growth cones. Together, these results suggest that netrin-1- and BDNF-induced growth cone attraction required ZBP1-mediated local translation of β-actin mRNA, and therefore ZBP1 regulates protein synthesis-dependent axon guidance. Thus, mRNA binding proteins regulating local translation can control spatiotemporal protein expression in response to guidance cues and directional cell motility.

Netrin-1-induced Local β-actin Synthesis and Growth Cone Guidance Requires Zipcode Binding Protein 1

RNA-binding proteins (RBPs) mediate the localization, stability, and translation of the target transcripts and fine-tune the physiological functions of the proteins encoded. The insulin-like growth factor (IGF) 2 mRNA-binding protein (IGF2BP, IMP) family comprises three RBPs, IGF2BP1, IGF2BP2, and IGF2BP3, capable of associating with IGF2 and other transcripts and mediating their processing. IGF2BP2 represents the least understood member of this family of RBPs; however, it has been reported to participate in a wide range of physiological processes, such as embryonic development, neuronal differentiation, and metabolism. Its dysregulation is associated with insulin resistance, diabetes, and carcinogenesis and may potentially be a powerful biomarker and candidate target for relevant diseases. This review summarizes the structural features, regulation, and functions of IGF2BP2 and their association with cancer and cancer stem cells.

/pdf/the-roles-of-insulin-like-growth-factor-2-mrna-binding-11u3fjn8cb.pdf

The Roles of Insulin-Like Growth Factor 2 mRNA-Binding Protein 2 in Cancer and Cancer Stem Cells

Fragile X Syndrome: from molecular pathology to therapy.

Intellectual disability (ID) and autism spectrum disorders (ADS) have in common alterations in some brain circuits and brain abnormalities, such as synaptic transmission and dendritic spines morphology. Recent studies have indicated a differential expression for specific categories of genes as a cause for both types of disease, while an increasing number of genes is recognized to produce both disorders. An example is the Fragile X Mental retardation gene, FMR1, whose silencing causes the Fragile X syndrome, the most common form of intellectual disability and autism, also characterized by physical hallmarks. FMRP, the protein encoded by FMR1, is an RNA-binding protein with an important role in translational control. Among the interactors of FMRP, CYFIP1/2 proteins are good candidates for intellectual disability and autism, on the bases of their genetic implication and functional properties, even if the precise functional significance of the CYFIP/FMRP interaction is not understood yet. CYFIP1 and CYFIP2 represent a link between Rac1, the Wave complex and FMRP, favoring the cross talk between actin polymerization and translational control

CYFIP family proteins between autism and intellectual disability: links with Fragile X syndrome

While innate behaviors are conserved throughout the animal kingdom, it is unknown whether common signaling pathways regulate the development of neuronal populations mediating these behaviors in diverse organisms. Here, we demonstrate that the Wnt/s-catenin effector Lef1 is required for the differentiation of anxiolytic hypothalamic neurons in zebrafish and mice, although the identity of Lef1-dependent genes and neurons differ between these 2 species. We further show that zebrafish and Drosophila have common Lef1-dependent gene expression in their respective neuroendocrine organs, consistent with a conserved pathway that has diverged in the mouse. Finally, orthologs of Lef1-dependent genes from both zebrafish and mouse show highly correlated hypothalamic expression in marmosets and humans, suggesting co-regulation of 2 parallel anxiolytic pathways in primates. These findings demonstrate that during evolution, a transcription factor can act through multiple mechanisms to generate a common behavioral output, and that Lef1 regulates circuit development that is fundamentally important for mediating anxiety in a wide variety of animal species.

/pdf/lef1-dependent-hypothalamic-neurogenesis-inhibits-anxiety-1h78pzguvh.pdf

Lef1-dependent hypothalamic neurogenesis inhibits anxiety

nev (cyfip2) is required for retinal lamination and axon guidance in the zebrafish retinotectal system

How neuronal connections are established during development is one of the most fascinating questions in the field of neurobiology. The zebrafish retinotectal system offers distinct advantages for studying axon guidance in an in vivo context. Its accessibility and the larva’s transparency not only allow its direct visualization, but also facilitate experimental manipulations to address the mechanisms of its development. Here we describe methods for labeling and visualizing retinal axons in vivo , including transient expression of DNA constructs, injection of lipophilic dyes, and time-lapse imaging. We describe in detail the available transgenic lines for marking retinal ganglion cells (RGCs); a protocol for very precise lipophilic dye labeling; and a protocol for single cell electroporation of RGCs. We then describe several approaches for perturbing the retinotectal system, including morpholino or DNA injection; localized heat shock to induce misexpression of genes; a comprehensive list of known retinotectal mutants; and a detailed protocol for RGC transplants to test cell autonomy. These methods not only provide new ways for examining how retinal axons are guided by their environment, but also can be used to study other axonal tracts in the living embryo.

Analyzing retinal axon guidance in zebrafish.

Attractive growth cone turning requires Igf2bp1-dependent local translation of β-actin mRNA in response to external cues in vitro. While in vivo studies have shown that Igf2bp1 is required for cell migration and axon terminal branching, a requirement for Igf2bp1 function during axon outgrowth has not been demonstrated. Using a timelapse assay in the zebrafish retinotectal system, we demonstrate that the β-actin 3’UTR is sufficient to target local translation of the photoconvertible fluorescent protein Kaede in growth cones of pathfinding retinal ganglion cells (RGCs) in vivo. Igf2bp1 knockdown reduced RGC axonal outgrowth and tectal coverage and retinal cell survival. RGC-specific expression of a phosphomimetic Igf2bp1 reduced the density of axonal projections in the optic tract while sparing RGCs, demonstrating for the first time that Igf2bp1 is required during axon outgrowth in vivo. Therefore, regulation of local translation mediated by Igf2bp proteins may be required at all stages of axon development.

/pdf/the-rna-binding-protein-igf2bp1-is-required-for-zebrafish-44ffoaa761.pdf

The RNA Binding Protein Igf2bp1 Is Required for Zebrafish RGC Axon Outgrowth In Vivo

Abstract Antagonistic interactions between center and surround receptive field (RF) components lie at the heart of the computations performed in the visual system. Circularly symmetric center-surround RFs are thought to enhance responses to spatial contrasts (i.e., edges), but how visual edges affect motion processing is unclear. Here, we addressed this question in retinal bipolar cells, the first visual neuron with classic center-surround interactions. We found that bipolar glutamate release emphasizes objects that emerge in the RF; their responses to continuous motion are smaller, slower, and cannot be predicted by signals elicited by stationary stimuli. In our hands, the alteration in signal dynamics induced by novel objects was more pronounced than edge enhancement and could be explained by priming of RF surround during continuous motion. These findings echo the salience of human visual perception and demonstrate an unappreciated capacity of the center-surround architecture to facilitate novel object detection and dynamic signal representation. 

/pdf/classical-center-surround-receptive-fields-facilitate-novel-2pkvt17g.pdf

John A. Gaynes

Papers

Lef1-dependent hypothalamic neurogenesis inhibits anxiety

nev (cyfip2) is required for retinal lamination and axon guidance in the zebrafish retinotectal system

Analyzing retinal axon guidance in zebrafish.

The RNA Binding Protein Igf2bp1 Is Required for Zebrafish RGC Axon Outgrowth In Vivo

Classical center-surround receptive fields facilitate novel object detection in retinal bipolar cells