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A Highly Reproducible and Straightforward Method to Perform <em>In Vivo</em> Ocular Enucleation in the Mouse after Eye Opening

Enucleation or the surgical removal of an eye can generally be considered as a model for nerve deafferentation. It provides a valuable tool to study the ...

KU Leuven - University of Leuven

Young adults were asked to solve two-digit addition problems and to say aloud the result of each calculation step to allow the identification of computation strategies. We manipulated the position of the largest addend (e.g., 25 + 48 vs. 48 + 25) to assess whether strategies are modulated by magnitude characteristics. With some strategies, participants demonstrated a clear preference to take the largest addend as the starting point for the calculation. Hence, rather than applying strategies in an inflexible manner, participants evaluated and compared the operands before proceeding to calculation. Further, mathematically skilled participants tended to use those magnitude-based strategies more often than less skilled ones. The findings demonstrate that magnitude information plays a role in complex arithmetic by guiding the process of strategy selection, and possibly more so for mathematically skilled participants.

Complex mental arithmetic: the contribution of the number sense.

Three experiments involving a Stroop-like paradigm were conducted. In Experiment 1, adults received a number comparison task in which large sets of dots, orthogonally varying along a discrete dimension (number of dots) and a continuous dimension (cumulative area), were presented. Incongruent trials were processed more slowly and with less accuracy than congruent trials, suggesting that continuous dimensions such as cumulative area are automatically processed and integrated during a discrete quantity judgement task. Experiment 2, in which adults were asked to perform area comparison on the same stimuli, revealed the reciprocal interference from number on the continuous quantity judgements. Experiment 3, in which participants received both the number and area comparison tasks, confirmed the results of Experiments 1 and 2. Contrasting with earlier statements, the results support the view that number acts as a more salient cue than continuous dimensions in adults. Furthermore, the individual predisposition to automatically access approximate number representations was found to correlate significantly with adults' exact arithmetical skills.

Judgement of discrete and continuous quantity in adults: number counts!

AMIGO2, or amphoterin-induced gene and ORF (open reading frame) 2, belongs to the leucine-rich repeats and immunoglobulin superfamilies. The protein is a downstream target of calcium-dependent survival signals and, therefore, promotes neuronal survival. Here, we describe the mRNA distribution pattern of AMIGO2 throughout the mouse brain with special emphasis on the hippocampus. In the Ammon's horn, a detailed comparison between the subregional mRNA expression patterns of AMIGO2 and Pcp4 (Purkinje cell protein 4)--a known molecular marker of hippocampal CA2 (Cornu Ammonis 2)--revealed a prominent AMIGO2 mRNA expression level in both the CA2 and the CA3a (Cornu Ammonis 3a) subregion of the dorsal and ventral hippocampus. Since this CA2/CA3a region is particularly resistant to neuronal injury and neurotoxicity [Stanfield and Cowan (Brain Res 309(2):299–307 1984); Sloviter (J Comp Neurol 280(2):183–196 1989); Leranth and Ribak (Exp Brain Res 85(1):129–136 1991); Young and Dragunow (Exp Neurol 133(2):125–137 1995); Ochiishi et al. (Neurosci 93(3):955–967 1999)], we suggest that the expression pattern of AMIGO2 indeed fits with its involvement in neuroprotection.

AMIGO2 mRNA expression in hippocampal CA2 and CA3a.

Counting and exact arithmetic rely on language-based representations, whereas number comparison and approximate arithmetic involve approximate quantity-based representations that are available early in life, before the first stages of language acquisition. The objective of this study was to examine the impact of language abilities on the later development of exact and approximate number skills.

Impact of language abilities on exact and approximate number skills development: evidence from children with specific language impairment.

Monocular enucleation (ME) drastically affects the contralateral visual cortex, where plasticity phenomena drive specific adaptations to compensate for the unilateral loss of vision. In adult mice, complete reactivation of deprived visual cortex involves an early visually driven recovery followed by multimodal plasticity 3 to 7 weeks post ME (Van Brussel et al. [2011] Cereb. Cortex 21:2133-2146). Here, we specifically investigated the age dependence of the onset and the exact timing of both ME-induced reactivation processes by comparing cortical activity patterns of mice enucleated at postnatal day (P) 45, 90, or 120. A swifter open-eye potentiated reactivation characterized the binocular visual cortex of P45 mice. Nevertheless, even after 7 weeks, the reactivation remained incomplete, especially in the monocular cortex medial to V1. In comparison with P45, emergent cross-modal participation was demonstrated in P90 animals, although robust reactivation similar to enucleated adults (P120) was not achieved yet. Concomitantly, at 7 weeks post ME, somatosensory and auditory cortex shifted from a hypoactive state in P45 to hyperactivity in P120. Thus, we provide evidence for a presensitive period in which gradual recruitment of cross-modal recovery upon long-term ME coincides with the transition from adolescence to adulthood in mice.

The cross-modal aspect of mouse visual cortex plasticity induced by monocular enucleation is age dependent.

It is largely admitted that processing numerosity relies on an innate Approximate Number System (ANS), and recent research consistently observed a relationship between ANS acuity and mathematical ability in childhood. However, studies assessing this relationship in adults led to contradictory results. In this study, adults with different levels of mathematical expertise performed two tasks on the same pairs of dot collections, based either on numerosity comparison or on cumulative area comparison. Number of dots and cumulative area were congruent in half of the stimuli, and incongruent in the other half. The results showed that adults with higher mathematical ability obtained lower Weber fractions in the numerical condition than participants with lower mathematical ability. Further, adults with lower mathematical ability were more affected by the interference of the continuous dimension in the numerical comparison task, whereas conversely higher-expertise adults showed stronger interference of the numerical dimension in the continuous comparison task. Finally, ANS acuity correlated with arithmetic performance. Taken together, the data suggest that individual differences in ANS acuity subsist in adulthood, and that they are related to mathematical ability.

Differences in the acuity of the Approximate Number System in adults: the effect of mathematical ability.

Although the Morris water maze (MWM) is the most frequently used protocol to examine hippocampus-dependent learning in mice, not much is known about the spatio-temporal dynamics of underlying plasticity processes. Here, we studied molecular and cellular hippocampal plasticity mechanisms during early and late phases of spatial learning in the MWM. Quantitative in situ hybridization for the immediate early genes zif268 and Homer1a (H1a) revealed phase-dependent differences in their expression between areas CA1 and CA3. During the initial learning phase, CA1 expression levels of the molecular plasticity marker H1a, but not of the activity reporter gene zif268, were related to task proficiency; whereas no learning-specific changes could be detected in CA3. Simultaneously, the ratio of surface-expressed NMDAR subunits NR2A and NR2B was downregulated as measured by acute slice biotinylation assay, while the total number of surface NMDARs was unaltered. When intrinsic 'somatic' and synaptic plasticity in the CA1-region of hippocampal slices were examined, we found that early learning promotes intrinsic neuronal plasticity as manifested by a reduction of spike frequency adaptation and postburst afterhyperpolarization. At the synaptic level, however, maintenance of long-term potentiation (LTP) in all learning groups was impaired which is most likely due to 'intrinsic' learning-induced LTP which occluded any further electrically induced LTP. Late learning, in contrast, was characterized by re-normalized H1a, NR2A and NR2B expression and neuronal firing, yet a further strengthening of learning-induced LTP. Together, our data support a precisely timed cascade of complex molecular and subcellular transformations occurring from early to late MWM learning.

Distinct and simultaneously active plasticity mechanisms in mouse hippocampus during different phases of Morris water maze training.

An ongoing debate in research on numerical cognition concerns the extent to which the approximate number system and symbolic number knowledge influence each other during development. The current study aims at establishing the direction of the developmental association between these two kinds of abilities at an early age. Fifty-seven children of 3-4 years performed two assessments at 7 months interval. In each assessment, children's precision in discriminating numerosities as well as their capacity to manipulate number words and Arabic digits was measured. By comparing relationships between pairs of measures across the two time points, we were able to assess the predictive direction of the link. Our data indicate that both cardinality proficiency and symbolic number knowledge predict later accuracy in numerosity comparison whereas the reverse links are not significant. The present findings are the first to provide longitudinal evidence that the early acquisition of symbolic numbers is an important precursor in the developmental refinement of the approximate number representation system.

Symbolic number abilities predict later approximate number system acuity in preschool children.

Matrix metalloproteinases (MMPs) are Zn(2+)-dependent endopeptidases considered to be essential for normal brain development and neuroplasticity by modulating extracellular matrix proteins, receptors, adhesion molecules, growth factors and cytoskeletal proteins. Specifically, MMP-3 has recently been implicated in synaptic plasticity, hippocampus-dependent learning and neuronal development and migration in the cerebellum. However, the function(s) of this enzyme in the neocortex is understudied. Therefore, we explored the phenotypical characteristics of the neuronal architecture and the capacity for experience-dependent cortical plasticity in the visual cortex of adult MMP-3-deficient (MMP-3(-/-)) mice. Golgi-Cox stainings revealed a significant reduction in apical dendritic length and an increased number of apical obliques for layer V pyramidal neurons in the visual cortex of adult MMP-3(-/-) mice compared to wild-type (WT) animals. In addition, a significant upregulation of both phosphorylated and non-phosphorylated neurofilament protein (NF)-high, phosphorylated NF-medium, NF-low and α-internexin was detected in the visual cortex of MMP-3(-/-) mice. To assess the effect of MMP-3 deficiency on cortical plasticity, we monocularly enucleated adult MMP-3(-/-) mice and analyzed the reactivation of the contralateral visual cortex 7 weeks post-enucleation. In contrast to previous results in C57Bl/6J adult mice, activity remained confined to the binocular zone and did not expand into the monocular regions indicative for an aberrant open-eye potentiation. Permanent hypoactivity in the monocular cortex lateral and medial to V1 also indicated a lack of cross-modal plasticity. These observations demonstrate that genetic inactivation of MMP-3 has profound effects on the structural integrity and plasticity response of the visual cortex of adult mice.

Julie Nys

Department of Biology

KU Leuven - University of Leuven

A Highly Reproducible and Straightforward Method to Perform In Vivo Ocular Enucleation in the Mouse after Eye Opening