Different grasping experiences affect mapping effects but not correspondence effects between stimulus size and response location.

The so-called spatial-size association of response codes (SSARC) effect denotes that humans respond faster and more accurately with a left response to physically small stimuli and a right response to physically large stimuli, as compared to the opposite mapping. According to an application of the CORE principle to the SSARC effect, the habit to grasp larger/heavier objects with one's dominant hand and smaller/lighter objects with one's non-dominant hand creates spatial-size associations. We investigated if grasping habits play a causal role in the formation of spatial-size associations by testing if the mapping of a preceding object-grasping task affects the size of the SSARC effect in subsequent choice-response tasks with keypress responses. In the object-grasping task, participants were instructed to grasp wooden cubes of variable size either according to a compatible (small-left; large-right) or according to an incompatible (small-right; large-left) mapping. In the choice-response tasks, participants responded with left or right keypresses to the size or color of a small or large stimulus. The results showed that participants with the compatible mapping in the object-grasping task showed a larger SSARC effect in the size discrimination task, but not in the color discrimination task, than participants with the incompatible mapping in the object-grasping task. Results suggest that a short period of practice with different size-location mappings can modulate size-location links used for controlled S-R translation, but not links underlying automatic S-R translation. In general, the results support the hypothesis that grasping habits play a causal role in the formation of spatial-size associations.

The reciprocity of spatial-numerical associations of vocal response codes depends on stimulus mode.

Individuals make faster left responses to small/er numbers and faster right responses to large/r numbers than vice versa. This "spatial-numerical association of response codes" (SNARC) effect represents evidence for an overlap between the cognitive representations of number and space. Theories of the SNARC effect differ in whether they predict bidirectional S-R associations between number and space or not. We investigated the reciprocity of S-R priming effects between number and location in three experiments with vocal responses. In Experiments 1 and 2, participants completed a number-location task, with digits as stimuli and location words as responses, and a location-number task, with physical locations as stimuli and number words as responses. In addition, we varied the S-R mapping in each task. Results revealed a strong SNARC effect in the number-location task, but no reciprocal SNARC effect in the location-number task. In Experiment 3, we replaced physical location stimuli with location words and digit stimuli with number words. Results revealed a regular and a reciprocal SNARC effect of similar size. Reciprocal SNARC effects thus seem to emerge with verbal location stimuli and vocal responses, but not with physical location stimuli and vocal responses. The S-R associations underlying the SNARC effect with vocal responses thus appear bidirectional and symmetrical for some combinations of stimulus and response sets, but not for others. This has implications for theoretical accounts of the SNARC effect which need to explain how stimulus mode affects the emergence of reciprocal but not regular SNARC effects.

TAOK2 rescues autism-linked developmental deficits in a 16p11.2 microdeletion mouse model.

The precise development of the neocortex is a prerequisite for higher cognitive and associative functions. Despite numerous advances that have been made in understanding neuronal differentiation and cortex development, our knowledge regarding the impact of specific genes associated with neurodevelopmental disorders on these processes is still limited. Here, we show that Taok2, which is encoded in humans within the autism spectrum disorder (ASD) susceptibility locus 16p11.2, is essential for neuronal migration. Overexpression of de novo mutations or rare variants from ASD patients disrupts neuronal migration in an isoform-specific manner. The mutated TAOK2α variants but not the TAOK2ß variants impaired neuronal migration. Moreover, the TAOK2α isoform colocalizes with microtubules. Consequently, neurons lacking Taok2 have unstable microtubules with reduced levels of acetylated tubulin and phosphorylated JNK1. Mice lacking Taok2 develop gross cortical and cortex layering abnormalities. Moreover, acute Taok2 downregulation or Taok2 knockout delayed the migration of upper-layer cortical neurons in mice, and the expression of a constitutively active form of JNK1 rescued these neuronal migration defects. Finally, we report that the brains of the Taok2 KO and 16p11.2 del Het mouse models show striking anatomical similarities and that the heterozygous 16p11.2 microdeletion mouse model displayed reduced levels of phosphorylated JNK1 and neuronal migration deficits, which were ameliorated upon the introduction of TAOK2α in cortical neurons and in the developing cortex of those mice. These results delineate the critical role of TAOK2 in cortical development and its contribution to neurodevelopmental disorders, including ASD.

Conserved Tao Kinase Activity Regulates Dendritic Arborization, Cytoskeletal Dynamics, and Sensory Function in Drosophila.

Dendritic arborization is highly regulated and requires tight control of dendritic growth, branching, cytoskeletal dynamics, and ion channel expression to ensure proper function. Abnormal dendritic development can result in altered network connectivity, which has been linked to neurodevelopmental disorders, including autism spectrum disorders (ASDs). How neuronal growth control programs tune dendritic arborization to ensure function is still not fully understood. Using Drosophila dendritic arborization (da) neurons as a model, we identified the conserved Ste20-like kinase Tao as a negative regulator of dendritic arborization. We show that Tao kinase activity regulates cytoskeletal dynamics and sensory channel localization required for proper sensory function in both male and female flies. We further provide evidence for functional conservation of Tao kinase, showing that its ASD-linked human ortholog, Tao kinase 2 (Taok2), could replace Drosophila Tao and rescue dendritic branching, dynamic microtubule alterations, and behavioral defects. However, several ASD-linked Taok2 variants displayed impaired rescue activity, suggesting that Tao/Taok2 mutations can disrupt sensory neuron development and function. Consistently, we show that Tao kinase activity is required in developing and as well as adult stages for maintaining normal dendritic arborization and sensory function to regulate escape and social behavior. Our data suggest an important role for Tao kinase signaling in cytoskeletal organization to maintain proper dendritic arborization and sensory function, providing a strong link between developmental sensory aberrations and behavioral abnormalities relevant for Taok2-dependent ASDs.SIGNIFICANCE STATEMENT Autism spectrum disorders (ASDs) are linked to abnormal dendritic arbors. However, the mechanisms of how dendritic arbors develop to promote functional and proper behavior are unclear. We identified Drosophila Tao kinase, the ortholog of the ASD risk gene Taok2, as a regulator of dendritic arborization in sensory neurons. We show that Tao kinase regulates cytoskeletal dynamics, controls sensory ion channel localization, and is required to maintain somatosensory function in vivo Interestingly, ASD-linked human Taok2 mutations rendered it nonfunctional, whereas its WT form could restore neuronal morphology and function in Drosophila lacking endogenous Tao. Our findings provide evidence for a conserved role of Tao kinase in dendritic development and function of sensory neurons, suggesting that aberrant sensory function might be a common feature of ASDs.

An anti-narcolepsy drug reveals behavioral and fitness costs of extreme activity cycles in arctic-breeding songbirds.

Sleep loss impairs cognitive function, immunological responses and general well-being in humans. However, sleep requirements in mammals and birds vary dramatically. In circumpolar regions with continuous summer light, daily sleep duration is reduced, particularly in breeding birds. The effect of an anti-narcolepsy drug (modafinil) to putatively extend wakefulness was examined in two species of closely related arctic-breeding passerine birds: Lapland longspurs (Calcarius lapponicus) and snow buntings (Plectrophenax nivalis). Free-living adult males were implanted during the nestling phase on day 4 (D4; 4 days post-hatching) with osmotic pumps containing either vehicle or modafinil to extend the active period for 72 h. Nestlings were weighed on D2 and D7 to measure growth rates. Additionally, focal observations were conducted on D6. Male longspurs receiving modafinil made fewer feeding visits and spent less time at the nest but tended to spend more time near the nest than controls. We observed no change in longspur nestling growth rates, but fledging occurred significantly later when males received modafinil, suggesting a fitness cost. In contrast, modafinil had no measurable impact on male or female snow bunting behavior, nestling growth rates or time to fledging. We suggest male longspurs compromise and maintain vigilance at their nests in lieu of sleeping because of the increased predation risk that is characteristic of their tundra nesting habitat. Snow buntings are cavity nesters, and their nests do not require the same vigilance, allowing males to presumably rest following provisioning. These life-history differences between species highlight the role of predation risk in mediating behavioral modifications to prolonged wakefulness in arctic-breeding songbirds.

Radial somatic F-actin organization affects growth cone dynamics during early neuronal development.

The centrosome is thought to be the major neuronal microtubule-organizing center (MTOC) in early neuronal development, producing microtubules with a radial organization. In addition, albeit in vitro, recent work showed that isolated centrosomes could serve as an actin-organizing center, raising the possibility that neuronal development may, in addition, require a centrosome-based actin radial organization. Here, we report, using super-resolution microscopy and live-cell imaging of cultured rodent neurons, F-actin organization around the centrosome with dynamic F-actin aster-like structures with F-actin fibers extending and retracting actively. Photoactivation/photoconversion experiments and molecular manipulations of F-actin stability reveal a robust flux of somatic F-actin toward the cell periphery. Finally, we show that somatic F-actin intermingles with centrosomal PCM-1 (pericentriolar material 1 protein) satellites. Knockdown of PCM-1 and disruption of centrosomal activity not only affect F-actin dynamics near the centrosome but also in distal growth cones. Collectively, the data show a radial F-actin organization during early neuronal development, which might be a cellular mechanism for providing peripheral regions with a fast and continuous source of actin polymers, hence sustaining initial neuronal development.

Altered TAOK2 activity causes autism-related neurodevelopmental and cognitive abnormalities through RhoA signaling.

Atypical brain connectivity is a major contributor to the pathophysiology of neurodevelopmental disorders (NDDs) including autism spectrum disorders (ASDs). TAOK2 is one of several genes in the 16p11.2 microdeletion region, but whether it contributes to NDDs is unknown. We performed behavioral analysis on Taok2 heterozygous (Het) and knockout (KO) mice and found gene dosage-dependent impairments in cognition, anxiety, and social interaction. Taok2 Het and KO mice also have dosage-dependent abnormalities in brain size and neural connectivity in multiple regions, deficits in cortical layering, dendrite and synapse formation, and reduced excitatory neurotransmission. Whole-genome and -exome sequencing of ASD families identified three de novo mutations in TAOK2 and functional analysis in mice and human cells revealed that all the mutations impair protein stability, but they differentially impact kinase activity, dendrite growth, and spine/synapse development. Mechanistically, loss of Taok2 activity causes a reduction in RhoA activation, and pharmacological enhancement of RhoA activity rescues synaptic phenotypes. Together, these data provide evidence that TAOK2 is a neurodevelopmental disorder risk gene and identify RhoA signaling as a mediator of TAOK2-dependent synaptic development.

Prospective observational cohort study of 'treatment as usual' over four years for patients with schizophrenia in a national forensic hospital.

BACKGROUND: We evaluated change in response to multi-modal psychosocial 'treatment as usual' programs offered within a forensic hospital. METHODS: Sixty nine patients with a diagnosis of schizophrenia or schizoaffective disorder were followed for up to four years. Patient progress was evaluated using the DUNDRUM-3, a measure of patient ability to participate and benefit from multi-modal psychosocial programs and the HCR-20 dynamic items, a measure of violence proneness. We report reliable change index (RCI) and reliable and clinically meaningful change (RMC). We assessed patients' cognition using the MCCB, psychopathology using the PANSS. The effect of cognition and psychopathology on change in DUNDRUM-3 was examined using hierarchical multiple regression with age, gender, and baseline DUNDRUM-3 scores. RESULTS: The DUNDRUM-3 changed significantly (p < 0.004, d = 0.367, RCI 32% of 69 cases, RMC 23%) and HCR-20-C (p < 0.003, d = 0.377, RCI 10%). Both cognition and psychopathology accounted for significant variance in DUNDRUM-3 at follow up. Those hospitalized for less than five years at baseline changed more than longer stay patients. Mediation analysis demonstrated that the relationship between cognition and change in violence proneness (HCR-20-C) was both directly affected and indirectly mediated by change in DUNDRUM-3. CONCLUSIONS: Change in response to multi-modal psychosocial programs (DUNDRUM-3) reduced a measure of violence proneness over four years. Forensic in-patients' ability to benefit from psychosocial treatment appears to be a function of the outcome measure used, unit of measurement employed, degree of cognitive impairment, psychopathology, and length of stay. Lower risk of re-offending may be partially attributable to participation and engagement in psychosocial interventions.

Sex-Dependent Phenological Plasticity in an Arctic Hibernator.

Hibernation provides a means of escaping the metabolic challenges associated with seasonality, yet the ability of mammals to prolong or reenter seasonal dormancy in response to extreme weather events is unclear. Here, we show that Arctic ground squirrels in northern Alaska exhibited sex-dependent plasticity in the physiology and phenology of hibernation in response to a series of late spring snowstorms in 2013 that resulted in the latest snowmelt on record. Females and nonreproductive males responded to the >1-month delay in snowmelt by extending heterothermy or reentering hibernation after several days of euthermy, leading to a >2-week delay in reproduction compared to surrounding years. In contrast, reproductive males neither extended nor reentered hibernation, likely because seasonal gonadal growth and development and subsequent testosterone release prevents a return to torpor. Our findings reveal intriguing differences in responses of males and females to climatic stressors, which can generate a phenological mismatch between the sexes.

Stable isotope analysis of CO2 in breath indicates metabolic fuel shifts in torpid arctic ground squirrels.

Stable carbon isotope ratios (δ13C) in breath show promise as an indicator of immediate metabolic fuel utilization in animals because tissue lipids have a lower δ13C value than carbohydrates and proteins. Metabolic fuel consumption is often estimated using the respiratory exchange ratio (RER), which has lipid and carbohydrate boundaries, but does not differentiate between protein and mixed fuel catabolism at intermediate values. Because lipids have relatively low δ13C values, measurements of stable carbon isotopes in breath may help distinguish between catabolism of protein and mixed fuel that includes lipid. We measured breath δ13C and RER concurrently in arctic ground squirrels (Urocitellus parryii) during steady-state torpor at ambient temperatures from -2 to -26°C. As predicted, we found a correlation between RER and breath δ13C values; however, the range of RER in this study did not reach intermediate levels to allow further resolution of metabolic substrate use with the addition of breath δ13C measurements. These data suggest that breath δ13C values are 1.1‰ lower than lipid tissue during pure lipid metabolism. From RER, we determined that arctic ground squirrels rely on nonlipid fuel sources for a significant portion of energy during torpor (up to 37%). The shift toward nonlipid fuel sources may be influenced by adiposity of the animals in addition to thermal challenge.

Handedness and effector strength modulate a compatibility effect between stimulus size and response position with manual and vocal responses.

Humans respond more quickly with the left hand to a small stimulus, and with the right hand to a large stimulus, as compared to the reverse mapping (spatial-size association of response codes [SSARC] effect). We investigated the hypothesis that strength differences between the hands contribute to the origin of this effect. Therefore, 80 left-handers and 80 right-handers participated in two experiments. In Experiment 1, participants performed a manual choice-response task in which we manipulated the mapping between physical stimulus size and responding hand. In addition, we measured the strengths of participants' left and right effectors (i.e., finger, hand, and arm). In Experiment 2, we measured the SSARC effect in vocal responses of the same sample. There were four main results. First, participants' dominant effectors were stronger than their nondominant effectors. Second, the SSARC effect occurred in manual and vocal responses with similar size. Third, in both modalities, the SSARC effect was larger in right-handers than in left-handers. Finally, strength differences between effectors (fingers and hands) correlated with the size of the SSARC effect. In sum, results support the hypothesis that functional differences between the hands contribute to the origin of the SSARC effect. In addition, the results suggest that size-space associations have generalized across motor systems, and formed a modality-independent association. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Protocol for differential multi-omic analyses of distinct cell types in the mouse cerebral cortex.

Here, we present a protocol for differential multi-omic analyses of distinct cell types in the developing mouse cerebral cortex. We describe steps for in utero electroporation, subsequent flow-cytometry-based isolation of developing mouse cortical cells, bulk RNA sequencing or quantitative liquid chromatography-tandem mass spectrometry, and bioinformatic analyses. This protocol can be applied to compare the proteomes and transcriptomes of developing mouse cortical cell populations after various manipulations (e.g., epigenetic). For complete details on the use and execution of this protocol, please refer to Meka et al. (2022).1.

The autism susceptibility kinase, TAOK2, phosphorylates eEF2 and modulates translation.

Genes implicated in translation control have been associated with autism spectrum disorders (ASDs). However, some important genetic causes of autism, including the 16p11.2 microdeletion, bear no obvious connection to translation. Here, we use proteomics, genetics, and translation assays in cultured cells and mouse brain to reveal altered translation mediated by loss of the kinase TAOK2 in 16p11.2 deletion models. We show that TAOK2 associates with the translational machinery and functions as a translational brake by phosphorylating eukaryotic elongation factor 2 (eEF2). Previously, all signal-mediated regulation of translation elongation via eEF2 phosphorylation was believed to be mediated by a single kinase, eEF2K. However, we show that TAOK2 can directly phosphorylate eEF2 on the same regulatory site, but functions independently of eEF2K signaling. Collectively, our results reveal an eEF2K-independent signaling pathway for control of translation elongation and suggest altered translation as a molecular component in the etiology of some forms of ASD.

Spatial-numerical associations of manual response codes are strongly asymmetrical.

The spatial-numerical association of response codes (SNARC) effect denotes the observation that humans respond faster and more accurately with a left-side response to smaller numbers and a right-side response to larger numbers, as compared to the opposite mapping. Existing accounts, such as the mental number line hypothesis or the polarity correspondence principle, differ in whether they assume symmetrical associations between numerical and spatial stimulus and response codes or not. In two experiments, we investigated the reciprocity of the SNARC effect in manual choice-response tasks with two conditions. In the number-location task, participants pressed a left or right key to a number stimulus (dots in Experiment 1, digits in Experiment 2). In the location-number task, participants made one or two consecutive keypresses with one hand to a left- or right-side stimulus. Both tasks were performed with a compatible (one-left, two-right; left-one, right-two) and an incompatible (one-right, two-left; left-two, right-one) mapping. In both experiments, results showed a strong compatibility effect in the number-location task, reflecting the typical SNARC effect. In contrast, in both experiments, there was no mapping effect in the location-number task when outliers were excluded. However, when outliers were not excluded, small reciprocal SNARC effects occurred in Experiment 2. Together, the findings suggest that priming of spatial responses by numerical stimuli is much stronger than priming of numerical responses by spatial stimuli. The results are consistent with some accounts of the SNARC effect (e.g., the mental number line hypothesis), but not with others (e.g., the polarity correspondence principle).

Associations between physical size and space are strongly asymmetrical.

The spatial-size association of response codes (SSARC) effect describes the phenomenon that left responses are faster and more accurate to small stimuli whereas right responses are faster and more accurate to large stimuli, as compared to the opposite mapping. The effect indicates associations between the mental representations of physical size and space. Importantly, the theoretical accounts of SSARC effects make different predictions about the reciprocity and/or symmetry of spatial-size associations. To investigate the reciprocity of SSARC effects, we compared compatibility effects in two verbal choice-response tasks: a size-location (typical SSARC) task and a location-size (reciprocal SSARC) task. In the size-location task, participants responded verbally to a small/large stimulus by saying "left"/"right". In the location-size task, participants responded verbally to a left-/right-side stimulus by saying "small"/"large". Participants completed both tasks with a compatible (small-left, large-right; left-small, right-large) and an incompatible (small-right, large-left; left-large, right-small) mapping. A regular SSARC effect emerged in the size-location task. However, no reciprocal SSARC effect emerged in the location-size task if outliers were excluded. If outliers were not excluded, small reciprocal SSARC effects occurred. Associations underlying the SSARC effect are thus strongly asymmetrical: Physical (stimulus) size can prime spatial responses much more strongly than spatial (stimulus) position can prime size-related responses. The finding of asymmetrical associations between size and space is in line with some theoretical accounts of the SSARC effect but at odds with others.

Exploring the response code in a compatibility effect between physical size and left/right responses: The hand is more important than location.

The spatial-size association of response codes (SSARC) effect refers to the finding of better performance with the left hand to small stimuli and with the right hand to large stimuli, as compared to the reverse mapping. In the present study, we investigated which response coding is responsible for the emergence of the SSARC effect. We observed a SSARC effect only with response selection between hands but not between fingers of one hand, indicating that the responses are coded relative to the body midline. Furthermore, we observed a SSARC effect with parallel arms but not with crossed arms, suggesting that both the anatomical side of the effector and its external spatial position contribute to the response code. However, using a reaching task as compared to keypresses, the SSARC effect followed the arms, suggesting that the crucial spatial response code refers more strongly to the anatomical side of the effector rather than to the external spatial response position. These findings document a strong influence of anatomically- or body-based coding on the SSARC effect, are at odds with the proposition of a generalized magnitude system that utilizes a common, external spatial metric, and point toward a categorical nature of response codes underlying the SSARC effect. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

The EphA4 receptor regulates dendritic spine remodeling by affecting beta1-integrin signaling pathways.

Remodeling of dendritic spines is believed to modulate the function of excitatory synapses. We previously reported that the EphA4 receptor tyrosine kinase regulates spine morphology in hippocampal pyramidal neurons, but the signaling pathways involved were not characterized (Murai, K.K., L.N. Nguyen, F. Irie, Y. Yamaguchi, and E.B. Pasquale. 2003. Nat. Neurosci. 6:153-160). In this study, we show that EphA4 activation by ephrin-A3 in hippocampal slices inhibits integrin downstream signaling pathways. EphA4 activation decreases tyrosine phosphorylation of the scaffolding protein Crk-associated substrate (Cas) and the tyrosine kinases focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (Pyk2) and also reduces the association of Cas with the Src family kinase Fyn and the adaptor Crk. Consistent with this, EphA4 inhibits beta1-integrin activity in neuronal cells. Supporting a functional role for beta1 integrin and Cas inactivation downstream of EphA4, the inhibition of integrin or Cas function induces spine morphological changes similar to those associated with EphA4 activation. Furthermore, preventing beta1-integrin inactivation blocks the effects of EphA4 on spines. Our results support a model in which EphA4 interferes with integrin signaling pathways that stabilize dendritic spines, thus modulating synaptic interactions with the extracellular environment.

Relative, not absolute, stimulus size is responsible for a correspondence effect between physical stimulus size and left/right responses.

Recent studies have demonstrated a novel compatibility (or correspondence) effect between physical stimulus size and horizontally aligned responses: Left-hand responses are shorter and more accurate to a small stimulus, compared to a large stimulus, whereas the opposite is true for right-hand responses. The present study investigated whether relative or absolute size is responsible for the effect. If relative size was important, a particular stimulus would elicit faster left-hand responses if the other stimuli in the set were larger, but the same stimulus would elicit a faster right-hand response if the other stimuli in the set were smaller. In terms of two-visual-systems theory, our study explores whether "vision for perception" (i.e., the ventral system) or "vision for action" (i.e., the dorsal system) dominates the processing of stimulus size in our task. In two experiments, participants performed a discrimination task in which they responded to stimulus color (Experiment 1) or to stimulus shape (Experiment 2) with their left/right hand. Stimulus size varied as an irrelevant stimulus feature, thus leading to corresponding (small-left; large-right) and non-corresponding (small-right; large-left) conditions. Moreover, a set of smaller stimuli and a set of larger stimuli, with both sets sharing an intermediately sized stimulus, were used in different conditions. The consistently significant two-way interaction between stimulus size and response location demonstrated the presence of the correspondence effect. The three-way interaction between stimulus size, response location, and stimulus set, however, was never significant. The results suggest that participants are inadvertently classifying stimuli according to relative size in a context-specific manner.

The Nature of Associations between Physical Stimulus Size and Left-Right Response Codes.

In two-choice response tasks, participants respond faster and more accurate with the left hand to a small stimulus and with the right hand to a large stimulus as compared to the reverse assignment. This compatibility effect suggests the existence of associations between cognitive codes of physical stimulus size and cognitive codes of left/right responses. Here, we explore the nature of associations between stimulus-size codes and left/right response codes by using more levels of stimulus size than in our previous studies. For example, the strengths of the associations between stimulus-size codes and response codes might either change gradually when stimulus size changes, or the strength of associations might change in a more discrete fashion (i.e., associations switch at a particular size level). In Experiment 1, participants responded to stimulus color with a left/right keypress, and physical stimulus size had ten levels with 5 mm steps. Results showed correspondence effects for the smallest and the largest stimulus size only. In Experiment 2, physical stimulus size had six levels with 10 mm steps. Results showed (similar) correspondence effects for the smallest and some intermediate stimulus-size levels. In sum, the results point towards a discrete, or categorical, relationship between cognitive codes of stimulus size and left/right response codes. This pattern of results is consistent with an account of the correspondence effect in terms of the polarity correspondence principle.

Pum2 and TDP-43 refine area-specific cytoarchitecture post-mitotically and modulate translation of Sox5, Bcl11b, and Rorb mRNAs in developing mouse neocortex.

In the neocortex, functionally distinct areas process specific types of information. Area identity is established by morphogens and transcriptional master regulators, but downstream mechanisms driving area-specific neuronal specification remain unclear. Here, we reveal a role for RNA-binding proteins in defining area-specific cytoarchitecture. Mice lacking Pum2 or overexpressing human TDP-43 show apparent 'motorization' of layers IV and V of primary somatosensory cortex (S1), characterized by dramatic expansion of cells co-expressing Sox5 and Bcl11b/Ctip2, a hallmark of subcerebral projection neurons, at the expense of cells expressing the layer IV neuronal marker Rorß. Moreover, retrograde labeling experiments with cholera toxin B in Pum2; Emx1-Cre and TDP43A315T mice revealed a corresponding increase in subcerebral connectivity of these neurons in S1. Intriguingly, other key features of somatosensory area identity are largely preserved, suggesting that Pum2 and TDP-43 may function in a downstream program, rather than controlling area identity per se. Transfection of primary neurons and in utero electroporation (IUE) suggest cell-autonomous and post-mitotic modulation of Sox5, Bcl11b/Ctip2, and Rorß levels. Mechanistically, we find that Pum2 and TDP-43 directly interact with and affect the translation of mRNAs encoding Sox5, Bcl11b/Ctip2, and Rorß. In contrast, effects on the levels of these mRNAs were not detectable in qRT-PCR or single-molecule fluorescent in situ hybridization assays, and we also did not detect effects on their splicing or polyadenylation patterns. Our results support the notion that post-transcriptional regulatory programs involving translational regulation and mediated by Pum2 and TDP-43 contribute to elaboration of area-specific neuronal identity and connectivity in the neocortex.