Two stages of programming eye gaze shifts in 3-D space.

Accurate saccadic and vergence eye movements towards selected visual targets are fundamental to perceive the 3-D environment. Despite this importance, shifts in eye gaze are not always perfect given that they are frequently followed by small corrective eye movements. The oculomotor system receives distinct information from various visual cues that may cause incongruity in the planning of a gaze shift. To test this idea, we analyzed eye movements in humans performing a saccade task in a 3-D setting. We show that saccades and vergence movements towards peripheral targets are guided by monocular (perceptual) cues. Approximately 200ms after the start of fixation at the perceived target, a fixational saccade corrected the eye positions to the physical target location. Our findings suggest that shifts in eye gaze occur in two phases; a large eye movement toward the perceived target location followed by a corrective saccade that directs the eyes to the physical target location.

Contextual modulation in V1: the Rossi-Zipser controversy.

A recent study (Rossi et al. 2001) implied the absence of figure-ground based contextual modulation in macaque V1, in contrast to the study on which it was modelled (Zipser et al. 1996). We suggest that Rossi et al. may have underestimated the extent of modulation by considering only positive and not also negative modulation and that their data may have shown figure-ground based contextual modulation. We then suggest a paradigm to assess whether it reflects figure-ground segregation, boundaries, or both.

The early differentiation of the neocortex: a hypothesis on neocortical evolution.

During development, a cerebral cortex appears in the wall of the telencephalic vesicle in reptiles and mammals. It arises from a cell-dense cortical plate, which develops within a primordial preplate. The neurons of the preplate are essential for cortical development; they regulate the neuronal migration of the cortical plate neurons and form the first axonal connections. In the reptilian cortex and in the hippocampus of the mammalian cerebral cortex, most ingrowing afferent axons run above the cortical plate, in the zone where the receptive tufts of apical dendrites of the cortical pyramidal neurons branch extensively. In the mammalian neocortex, however, axons enter mainly from below the cortical plate where they do not encounter the apical tufts of these pyramidal neurons. In this paper, we discuss the idea that this difference in cortical development has relieved a functional constraint in the expansion of the cortex during evolution. We hypothesize that the entrance of axons below the cell-dense cortical plate, together with the inside-out migration of cortical neurons, ensures that the neocortex remains an "open" system, able to differentiate into new (sub)layers and more cortical areas.

Chromosomal instability in chromosome band 12p13: multiple breaks leading to complex rearrangements including cytogenetically undetectable sub-clones.

During fluorescence in situ hybridization (FISH) analysis of metaphase cells from 70 patients with lymphoid and myeloid hematologic malignancies and chromosomal rearrangements involving band 12p13, we identified nine patients (four with lymphoid malignancies, four with myeloid malignancies and one with biphenotypic leukemia) who showed more complicated rearrangements than we had expected from conventional cytogenetic study. In six patients, multiple breaks occurred in small segments of 12p with subsequent translocations and insertions of these segments into other chromosomes, sometimes to unexpected regions. In three patients additional chromosome breaks resulted in a sub-clone which was cytogenetically indistinguishable from the main clone in each patient based on the cytogenetic analysis. These subtle molecular events were detected exclusively in a region covering TEL/ETV6 and KIP1/CDKN1B. Seven of nine had a previous history of chemo/radiotherapy; all the patients showed complex karyotypes, even though they were newly diagnosed with leukemia. Survival data were available in five patients, and all survived less than 6 months. These findings suggest that the 12p13 region, especially the above-mentioned region, is genetically unstable and fragile. It is likely that multiple chromosome breaks were induced through mutagens used in chemo/ radiotherapy, and are associated with a sub-group of patients with an extremely bad prognosis.

A neural correlate of working memory in the monkey primary visual cortex.

The brain frequently needs to store information for short periods. In vision, this means that the perceptual correlate of a stimulus has to be maintained temporally once the stimulus has been removed from the visual scene. However, it is not known how the visual system transfers sensory information into a memory component. Here, we identify a neural correlate of working memory in the monkey primary visual cortex (V1). We propose that this component may link sensory activity with memory activity.

Two distinct modes of sensory processing observed in monkey primary visual cortex (V1).

Even salient sensory stimuli are sometimes not detected. What goes wrong in the brain in that case? Here we show that a late (> 100-ms) component of the neural activity in the primary visual cortex of the monkey is selectively suppressed when stimuli are not seen. As there is evidence that this activity depends on feedback from extrastriate areas, these findings suggest a specific role for recurrent processing when stimuli are reaching a perceptual level. Further results show that this perceptual level is situated between purely sensory and decision or motor stages of processing.

Disruption of neuronal migration and radial glia in the developing cerebral cortex following ablation of Cajal-Retzius cells.

Cortical neurons are generated within the proliferative layer and follow a strict 'inside-out' gradient of migration and positioning, which determines the characteristic layering and pattern of neural connections in the adult cerebral cortex. Thus, directional migration of postmitotic neuroblasts towards layer I and regulation of the radial glia phenotype subserving cortical migration are central issues in corticogenesis. Recent studies showing that the gene disrupted in the reeler mutation--reelin--is expressed in Cajal-Retzius cells have indicated a role for these pioneer neurons in cortical migration. We show here that ablation of Cajal-Retzius cells in layer I by local application of domoic acid in newborn mice arrests migration of the late-generated neurons, destined to cortical layers II-III, that have been labeled by 5-bromodeoxyuridine injections administered at E16. In addition, degeneration of Cajal-Retzius cells in newborn mice dramatically decreases the number of radial glial apical processes identified by nestin-immunostaining, but increases the number of maturing glial fibrillary acidic protein-positive astrocytes. These findings support an essential role for Cajal-Retzius cells in neuronal migration and corticogenesis, by regulating the identity and function of radial glia and the radial glia-to-astrocyte transformation.

The role of primary visual cortex (V1) in visual awareness.

In the search for the neural correlate of visual awareness, much controversy exists about the role of primary visual cortex. Here, the neurophysiological data from V1 recordings in awake monkeys are examined in light of two general classes of models of visual awareness. In the first model type, visual awareness is seen as being mediated either by a particular set of areas or pathways, or alternatively by a specific set of neurons. In these models, the role of V1 seems rather limited, as the mere activity of V1 cells seems insufficient to mediate awareness. In the second model type, awareness is hypothesized to be mediated by a global mechanism, i.e. a specific kind of activity not linked to a particular area or cell type. Two separate versions of global models are discussed, synchronous oscillations and spike rate modulations. It is shown that V1 synchrony does not reflect perception but rather the horizontal connections between neurons, indicating that V1 synchrony cannot be a direct neural correlate of conscious percepts. However, the rate of spike discharges of V1 neurons is strongly modulated by perceptual context, and these modulations correlate very well with aspects of perceptual organization, visual awareness, and attention. If these modulations serve as a neural correlate of visual awareness, then V1 contributes to that neural correlate. Whether V1 plays a role in the neural correlate of visual awareness thus strongly depends on the way visual awareness is hypothesized to be implemented in the brain.

The early development of thalamocortical and corticothalamic projections in the mouse.

The initial ingrowth of corticothalamic and thalamocortical projections was examined in mice at embryonic and perinatal stages. Fibers, in fixed brains, were labeled with the carbocyanine dye 1, 1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocianine perchlorate (DiI). By E13, the corticofugal fibers had entered the lowest intermediate zone through which they ran, turned over the corpus striatum, and left the cortex. The fibers were arranged in scattered bundles throughout the corpus striatum. At E14 corticofugal axons reached the internal capsule and at E14.5-E15 they established contact within the thalamus. Meanwhile, the thalamocortical afferents reached the neocortex at E13. At this time fibers ran tangentially within the intermediate zone, immediately underneath the cortical plate. By E14, the fibers had started to invade the subplate and, by E15, thalamocortical fibers had begun their radial growth into the cortex. Such radial growth proceeded steadily, invading each cortical layer as it differentiated cytoarchitectonically from the dense cortical plate. The first retrogradely labeled cells were detected at the cortical plate at E15. By the day of birth (E20), thalamocortical fibers had formed a dense branching system within layers VI and V. Our observations indicate that, in mice, the thalamic axons reach the cortex before corticothalamic projections enter the thalamic nuclei. Moreover, the results suggest that the pathway followed by each fiber system is different. By DiI injections into the internal capsule we have also determined that subplate cells are the first to send axons to the thalamus.

Fine mapping of the friend retrovirus resistance gene, Rfv3, on mouse chromosome 15.

Rfv3 is a host resistance gene that operates through an unknown mechanism to control the development of the virus-neutralizing antibody response required for recovery from infection with Friend retrovirus. The Rfv3 gene was previously mapped to an approximately 20-centimorgan (cM) region of chromosome 15. More refined mapping was not possible, due to a lack of microsatellite markers and leakiness in the Rfv3 phenotype, which prevented definitive phenotyping of individual recombinant mice. In the present study, we overcame these difficulties by taking advantage of seven new microsatellite markers in the Rfv3 region and by using progeny tests to accurately determine the Rfv3 phenotype of recombinant mice. Detailed linkage analysis of relevant crossovers narrowed the location of Rfv3 to a 0.83-cM region. Mapping of closely linked genes in an interspecific backcross panel allowed us to exclude two previous candidate genes, Ly6 and Wnt7b. These studies also showed for the first time that the Hsf1 gene maps to the Rfv3-linked cluster of genes including Il2rb, Il3rb, and Pdgfb. This localization of Rfv3 to a region of less than 1 cM now makes it feasible to attempt the cloning of Rfv3 by physical methods.

Requirement for CD4(+) T cells in the Friend murine retrovirus neutralizing antibody response: evidence for functional T cells in genetic low-recovery mice.

Recovery from infection with the Friend murine leukemia retrovirus complex (FV) requires T-helper cells and cytotoxic T cells as well as neutralizing antibodies. Several host genes, including genes of the major histocompatibility complex (H-2) and an H-2-unlinked gene, Rfv-3, influence these FV-specific immune responses. (B10.A x A/Wy)F1 mice, which have the H-2(a/a) Rfv-3(r/s) genotype, fail to mount a detectable FV-specific T-cell proliferative response but nevertheless produce FV-specific neutralizing immunoglobulin M (IgM) antibodies and can eliminate FV viremia. Thus, this IgM response, primarily influenced by the Rfv-3 gene, may be T-cell independent. To test this idea, mice were depleted of either CD4(+) or CD8(+) T-cell populations in vivo and were monitored for the effect on the neutralizing antibody response following FV infection. Surprisingly, mice in which CD4(+) cells were depleted showed undetectable FV-neutralizing antibody responses and high viremia levels compared to nondepleted or CD8-depleted animals. In addition to knocking out the FV antibody response, CD4(+) T-cell depletion reduced survival time significantly, further indicating the importance of CD4(+) T cells. These studies revealed the first evidence for a functional T-cell response following FV infection in these low-recovery mice and showed that CD4(+) T-helper cells are required for the Rfv-3-controlled FV antibody response.

Feedforward, horizontal, and feedback processing in the visual cortex.

The cortical visual system consists of many richly interconnected areas. Each area is characterized by more or less specific receptive field tuning properties. However, these tuning properties reflect only a subset of the interactions that occur within and between areas. Neuronal responses may be modulated by perceptual context or attention. These modulations reflect lateral interactions within areas and feedback from higher to lower areas. Recent work is beginning to unravel how horizontal and feedback connections each contribute to modulatory effects and what the role of these modulations is in vision. Whereas receptive field tuning properties reflect feedforward processing, modulations evoked by horizontal and feedback connections may reflect the integration of information that underlies perception.

The functions of the preplate in development and evolution of the neocortex and hippocampus.

Recently, it has been shown that the early developmental organization of the archicortical hippocampus resembles that of the neocortex. In both cortices at embryonic stages, a preplate is present, which is split by the formation of the cortical plate into a marginal zone and a subplate layer. The pioneer neurons of the preplate are believed to form a phylogenetically ancient cortical structure. Neurons in these preplate layers are the first postmitotic neurons and have important roles in the development of the cerebral cortex. Cajal-Retzius cells in the marginal zone regulate the phenotype of radial glial cells and may direct neuronal migration establishing the inside-out gradient of corticogenesis. Furthermore, pioneer neurons form the initial axonal connections with other (sub)cortical structures. A significant difference between the hippocampus and neocortex, however, is that in the hippocampus, most afferents are guided by the pioneer neurons in the prominent marginal zone, while in the neocortex most ingrowing afferent axons enter via the subplate. At later developmental periods, most pioneer neurons disappear by cell death or transform into other neuronal shapes. Here, we review the early developmental organization of the mammalian cerebral cortex (both neocortex and hippocampus) and discuss the functions and fate of pioneer neurons in cortical development, in particular that of Cajal-Retzius cells. Evaluating the developmental properties of the hippocampus and neocortex, we present the hypothesis that the distribution of the main ingrowing afferent systems in the developing neocortex, which differs from the one in the hippocampal region, may have enabled the specific evolution of the neocortex.

Involvement of distinct pioneer neurons in the formation of layer-specific connections in the hippocampus.

During neural development, specific recognition molecules provide the cues necessary for the formation of initial projection maps, which are reshaped later in development. In some systems, guiding cues for axonal pathfinding and target selection are provided by specific cells that are present only at critical times. For instance, the floor plate guides commissural axons in the spinal cord, and the subplate is involved in the formation of thalamocortical connections. Here we study the development of entorhinal and commissural connections to the murine hippocampus, which in the adult terminate in nonoverlapping layers. We show that two groups of pioneer neurons, Cajal-Retzius cells and GABAergic neurons, form layer-specific scaffolds that overlap with distinct hippocampal afferents at embryonic and early postnatal stages. Furthermore, at postnatal day 0 (P0)-P5, before the dendrites of pyramidal neurons develop, these pioneer neurons are preferential synaptic targets for hippocampal afferents. Birthdating analysis using 5'-bromodeoxyuridine (BrdU) pulses showed that most such early-generated neurons disappear at late postnatal stages, most likely by cell death. Together with previous studies, these findings indicate that distinct pioneer neurons are involved in the guidance and targeting of different hippocampal afferents.

Identification of complex genomic breakpoint junctions in the t(9;11) MLL-AF9 fusion gene in acute leukemia.

The MLL gene at chromosome 11, band q23, is involved in translocations with as many as 40 different chromosomal bands. Virtually all breakpoints occur within an 8.3 kb BamHI fragment and result in 5' MLL fused to partner genes in a 5'-3' orientation. The translocation t(9;11)(p22;q23), which results in the fusion of MLL to AF9, is the most common of the 11q23 chromosomal abnormalities observed in de novo acute myeloid leukemia (AML), in therapy related leukemia (t-AML), and rarely in acute lymphoblastic leukemia (ALL). We have studied 24 patients with a t(9;11) and an MLL rearrangement, including 19 patients with AML, four with t-AML, and one with ALL. To understand the mechanisms of this illegitimate recombination, we cloned and sequenced the t(9;11) translocation breakpoint junctions on both derivative chromosomes from one AML patient and from the Mono Mac 6 (MM6) cell line, which was derived from a patient with AML. Two different complex junctions were noted. In the AML patient, both chromosome 11 and 9 breaks were staggered, occurred in Alu DNA sequences, and resulted in a 331 bp duplication. In the MM6 cell line, breaks in chromosomes 11 and 9 were also staggered, but, in contrast to the finding in the AML patient, the breaks did not involve Alu DNA sequences and resulted in a 664 bp deletion at the breakpoints. Using reverse transcriptase (RT-) PCR, we analyzed 11 patient samples, including the two just described, for MML-AF9 fusions. The fusion occurred in six of seven AML patients, two of two t-AML patients, one patient with ALL, and in the MM6 cell line. Interestingly, all of the breaks within the AF9 gene in AML patients occurred in the central AF9 exon, called Site A by others, whereas in the single ALL patient the breakpoint mapped to a more 3' region of the AF9 gene. Our data, when combined with those of others, suggest that the fusion point within the AF9 gene, and thus the amount of AF9 material included in the MLL-AF9 fusion gene product, may influence the phenotype of the resulting leukemia. This further supports the proposal that the MML translocation partner genes play a critical role in the leukemogenic process.

Survival of Cajal-Retzius cells after cortical lesions in newborn mice: a possible role for Cajal-Retzius cells in brain repair.

Transient Cajal-Retzius (CR) cells in layer I of the mammalian cerebral cortex are the first postmitotic neurons and they are believed to play a role in neuronal migration and lamination during cortical development. Freezing insults to the cortex of newborn mice produce cortical malformations similar to those observed in human brain disorders. Here we have used calretinin immunostaining to investigate the response of CR cells to freezing lesions of the cortical surface. Shortly after injury, CR cells disappeared from the lesioned zone. Moreover, CR cells located near the lesioned area adopted extremely fusiform shapes. At later postnatal stages (P12), CR cells were still abundant in layer I of the lesioned zone, in contrast to their almost complete loss in control animals. These results show that CR cells survive for longer developmental periods following cortical injury. Furthermore, the initial loss and later re-appearance of CR cells suggest that these neurons might migrate tangentially from the cortical areas surrounding the lesioned zone. These findings imply a role for CR cells in brain repair after cortical injury during development.

Degeneration of Cajal-Retzius cells in the developing cerebral cortex of the mouse after ablation of meningeal cells by 6-hydroxydopamine.

In the central nervous system, the neurotoxic drug 6-hydroxydopamine (6-OHDA) selectively deletes central catecholaminergic neurons and meningeal cells. Meningeal cells are known to contribute to brain development and their specific degeneration leads to disorganized neuronal positioning. We have analyzed whether a particular population of cortical pioneer neurons, the Cajal-Retzius (CR) cells, which lie just below meningeal cells, is also affected by 6-OHDA treatment. We show that application of 6-OHDA to the cortical surface leads to a rapid degeneration of CR cells, without affecting other cortical neurons. The ablation of CR cells was prevented by normetanephrine, which blocks the 6-OHDA uptake into meningeal cells. These results indicate that the disappearance of CR cells after 6-OHDA treatment may be a result of the ablation of the meningeal cells and suggest a trophic dependence of CR cells upon meningeal cells.

Differential survival of Cajal-Retzius cells in organotypic cultures of hippocampus and neocortex.

Cajal-Retzius (CR) cells are transient, pioneer neurons of layer I of the cortex that are believed to play essential roles in corticogenesis, e.g., in neuronal migration and synaptogenesis. Here we have used calretinin immunostaining to study the characteristics, survival, and fate of CR cells in single organotypic slice cultures of mouse neocortex and hippocampus deprived of their extrinsic afferents. In neocortical explants, CR cells were observed after 1-3 d in vitro (DIV), but they disappeared after 5-7 DIV, which is similar to their time of degeneration in vivo. The disappearance of CR cells in neocortical slices was prevented by incubation with tetrodotoxin and the glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3,-dione but not by 2-amino-5-phosphonopentanoic acid, suggesting that neuronal activity and non-NMDA glutamate receptors may trigger CR cell death in the neocortex. In contrast to the situation in vivo, in which many hippocampal CR cells disappear at approximately the third postnatal week, CR cells survived in single hippocampal cultures after long incubation times (31 DIV), with their morphology essentially unaltered. In contrast, fewer CR cells were found when hippocampal slices were cocultured with explants from the entorhinal cortex. Because CR cells are transient synaptic targets for entorhinohippocampal afferents, these findings suggest a role for entorhinal afferents in the degeneration of CR cells in the hippocampus. In conclusion, this study shows different survival properties of CR cells in organotypic slice cultures of hippocampus and neocortex, and it suggests that different mechanisms are involved in the regulation of the process of naturally occurring CR cell death in the two cortical regions.

Distribution of 11q23 breakpoints within the MLL breakpoint cluster region in de novo acute leukemia and in treatment-related acute myeloid leukemia: correlation with scaffold attachment regions and topoisomerase II consensus binding sites.

A major unresolved question for 11q23 translocations involving MLL is the chromosomal mechanism(s) leading to these translocations. We have mapped breakpoints within the 8.3-kb BamHI breakpoint cluster region in 31 patients with acute lymphoblastic leukemia and acute myeloid leukemia (AML) de novo and in 8 t-AML patients. In 23 of 31 leukemia de novo patients, MLL breakpoints mapped to the centromeric half (4.57 kb) of the breakpoint cluster region, whereas those in eight de novo patients mapped to the telomeric half (3.87 kb). In contrast, only two t-AML breakpoints mapped in the centromeric half, whereas six mapped in the telomeric half. The difference in distribution of the leukemia de novo breakpoints is statistically significant (P = .02). A similar difference in distribution of breakpoints between de novo patients and t-AML patients has been reported by others. We identified a low- or weak-affinity scaffold attachment region (SAR) mapping just centromeric to the breakpoint cluster region, and a high-affinity SAR mapping within the telomeric half of the breakpoint cluster region. Using high stringency criteria to define in vitro vertebrate topoisomerase II (topo II) consensus sites, one topo II site mapped adjacent to the telomeric SAR, whereas six mapped within the SAR. Therefore, 74% of leukemia de novo and 25% of t-AML breakpoints map to the centromeric half of the breakpoint cluster region map between the two SARs; in contrast, 26% of the leukemia de novo and 75% of the t-AML patient breakpoints map to the telomeric half of the breakpoint cluster region that contains both the telomeric SAR and the topo II sites. Thus, the chromatin structure of the MLL breakpoint cluster region may be important in determining the distribution of the breakpoints. The data suggest that the mechanism(s) leading to translocations may differ in leukemia de novo and in t-AML.

Migrating neurons in the developing cerebral cortex of the mouse send callosal axons.

The presence of migrating callosal neurons during the development of the murine cerebral cortex was studied using biocytin and the lipophilic dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate as retrograde tracers. After injections of biocytin in the presumptive somatosensory cortex of newborn mice which were analysed one day later, many anterogradely labelled fibres coursed towards the contralateral hemisphere through the corpus callosum. Retrogradely labelled callosal cells were also observed. Most callosal neurons corresponded to immature pyramidal cells. In addition, a few biocytin-labelled callosal neurons displayed extremely fusiform shapes, vertical orientation and a short, single process emerging from the apical side of the perikaryon. At the electron microscopic level, these cells had features identical to those described for migrating callosal neurons. Twenty-four hours after birth, these migrating neurons were almost exclusively observed in the upper, dense aspect of the cortical plate (presumptive layers II-III) and only very exceptionally in the infragranular layers. No retrogradely labelled cell resembling migrating neurons were noticed after injections on postnatal days 2 or 5. To study migrating callosal neurons at embryonic stages, crystals of the lipophilic dye were injected in the corpus callosum or the contralateral white matter in embryos aged 17, 18 and 19 days, corresponding to the initial development of the corpus callosum in mice. Whereas callosal migrating neurons were not detected at embryonic days 17 and 18, injections of the lipophilic dye on embryonic day 19 revealed the presence of labelled migrating neurons in the infragranular layers. To corroborate further that these cells are migrating neurons, [3H]thymidine was administered on embryonic days 16 and 17, and labelled mice were injected with biocytin on embryonic day 19 or the first postnatal day. Retrogradely labelled callosal neurons resembling migrating neurons were autoradiographically labelled. These results indicate that the specification of certain neuronal types and the emergence of their cell type-specific characteristics occur shortly after postmitotic neurons leave the ventricular zone, before being positioned within the cortical plate.