Switch time-point for rapid experience-dependent changes in zinc-containing circuits in the mouse barrel cortex

It has been previously demonstrated that in the mouse barrel cortex, synaptic zinc is regulated by sensory experience. In adult mice, cutting selected vibrissae produced a rapid but transient elevation of synaptic zinc in the corresponding barrels several hours later, whereas in 8 day-old animals this procedure did not affect synaptic zinc. In the present study, we wished to determine the postnatal age at which zinc-containing terminals gain the ability to respond rapidly to a restriction of sensory input. We therefore examined the effects of 1-day sensory deprivation starting at different postnatal ages. For this purpose we unilaterally trimmed all rows of vibrissae, except for row C, and we then visualized synaptic zinc in the barrel cortex 24h later. Up to postnatal day 15 such procedure had no effect on the level of synaptic zinc. However, beginning at postnatal day 16, 1-day sensory deprivation produced an increase in synaptic zinc within hollows of deprived rows of barrels as compared to non-deprived rows. These results show that during development there is a specific time-point after which zinc-containing circuits may respond rapidly to altered sensory inputs. A comparison of these findings with previous results obtained after chronic sensory deprivation suggests that a specific time window exists in development for persistent alterations in zinc-containing circuits.     

Experience-dependent plasticity of zinc-containing cortical circuits during a critical period of postnatal development

Distinctive subsets of glutamatergic neurons in cerebral cortex sequester the transition metal zinc within the synaptic vesicles of their axon terminals. In the present study we used histochemical localization of synaptic zinc to investigate normal postnatal development and experience-dependent plasticity of zinc-containing circuits in somatosensory barrel cortex of rats. First, we found that zinc-containing cortical circuits are dynamically reorganized between postnatal day (P) 0 and P28. Whereas most cortical laminae exhibited idiosyncratic increases in zinc histochemical staining with advancing age, lamina IV barrels were darkly reactive early in life and then lost much of their complement of synaptic zinc during postnatal weeks 2-4. Second, we established that sensory experience plays a major role in sculpting the zinc-containing innervation of cortical barrels. Trimming a particular facial whisker arrested the normal postnatal decline in synaptic zinc in the corresponding, deprived barrel. This resulted in more intense zinc staining in deprived barrels compared with adjacent, nondeprived barrels. Notably, the influence of experience on development of zinc circuits was most robust during a critical period extending from about P14, when an effect of whisker trimming first could be observed, through P28, after which time chronic deprivation no longer resulted in heightened levels of synaptic zinc in lamina IV. These findings indicate that sensory input can have a marked influence on development of cortical circuits, including those within lamina IV, throughout the first postnatal month.     

Experience-dependent structural plasticity in the cortex

Synapses are the fundamental units of neuronal circuits. Synaptic plasticity can occur through changes in synaptic strength, as well as through the addition/removal of synapses. Two-photon microscopy in combination with fluorescence labeling offers a powerful tool to peek into the living brain and follow structural reorganization at individual synapses. Time-lapse imaging depicts a dynamic picture in which experience-dependent plasticity of synaptic structures varies between different cortical regions and layers, as well as between neuronal subtypes. Recent studies have demonstrated that the formation and elimination of synaptic structures happens rapidly in a subpopulation of cortical neurons during various sensorimotor learning experiences, and that stabilized synaptic structures are associated with long lasting memories for the task. Therefore, circuit plasticity, mediated by structural remodeling, provides an underlying mechanism for learning and memory.     

Circadian rhythmicity of synapses in mouse somatosensory cortex

The circadian rhythmicity displayed by motor behavior of mice: activity at night and rest during the day; and the associated changes in the sensory input are reflected by cyclic synaptic plasticity in the whisker representations located in the somatosensory (barrel) cortex. It was not clear whether diurnal rhythmic changes in synapse density previously observed in the barrel cortex resulted from changes in the activity of the animals, from daily light/dark (LD) rhythm or are driven by an endogenous clock. These changes were investigated in the barrel cortex of C57BL/6 mouse strain kept under LD 12 : 12 h conditions and in constant darkness (DD). Stereological analysis of serial electron microscopic sections was used to assess numerical density of synapses. In mice kept under LD conditions, the total density of synapses and the density of excitatory synapses located on dendritic spines was higher during the light period (rest phase). In contrast, the density of inhibitory synapses located on dendritic spines increased during the dark period (activity phase). Under DD conditions, the upregulation of the inhibitory synapses during the activity phase was retained, but the cyclic changes in the density of excitatory synapses were not observed. The results show that the circadian plasticity concerns only synapses located on spines (and not those on dendritic shafts), and that excitatory and inhibitory synapses are differently regulated during the 24 h cycle: the excitatory synapses are influenced by light, whilst the inhibitory synapses are driven by the endogenous circadian clock.     

Paradoxical effects of minocycline in the developing mouse somatosensory cortex

Minocycline, a tetracycline derivative, is known to exert neuroprotective effects unrelated to its antimicrobial action. In particular, minocycline prevents microglial activation in pathological conditions and consequently reduces the production of proinflammatory factors contributing to the propagation of diseases. Accumulative evidence indicates that microglial cells contribute to the maturation of neuronal and synaptic networks during the normal development of the central nervous system (CNS) and that perinatal inflammation is a known risk factor for brain lesions. Although minocycline has been used to infer microglia functions during development, mechanisms by which this tetracycline derivative affect the immature CNS have not been analyzed in detail. In this study, we demonstrate that minocycline administration during the first postnatal week of development has paradoxical effects on microglia phenotype and on neuronal survival in the mouse somatosensory cortex. Using a combination of immunohistochemistry and electrophysiology, we show that intraperitoneal injections of minocycline between postnatal days 6 and 8 affect distribution, morphology, and functional properties of microglia cells of the whisker‐related barrel cortex, leading to the development of a phenotype resembling that of microglia activated in pathological conditions. Minocyline also induced a massive cell death that developed faster than changes in microglia phenotype, suggesting that the latter is a consequence of the former. Finally, cell death and microglial activation were not observed when minocycline treatment was postponed by only 2 days (i.e., between postnatal days 8 and 10). These observations call into question the use of tetracycline derivatives during CNS development to study microglia or to reduce perinatal inflammation. GLIA 2014;62:399–410     

Distribution of synaptic zinc in the developing mouse somatosensory barrel cortex

Histochemical localization of synaptic zinc was examined in the somatosensory (SI) barrel cortex of mouse. The laminar distribution and distribution within the barrel field were described. At postnatal day 3 (P3) and 5 (P5), very faint and uniform zinc staining was present in the lower part of the subplate. At P6, subtle laminar variations emerged. At P8, these variations were clearly observed. Intense zinc staining was found in layers I, II, III, and V. Layers IV and VI showed a weaker staining. From this postnatal age to adult, uneven patchy distribution of synaptic zinc in layer IV could be distinguished in coronal sections. In tangential sections through layer IV, zinc staining showed a barrel-like pattern due to a higher zinc concentration in septa and the surrounding cortex. Barrel sides revealed a lower zinc concentration compared with the barrel hollow. With brain maturation, the zinc staining increased more intensely outside the barrel field, thus producing a progressively higher contrast between the barrel field and adjacent cortical regions. The differences in zinc staining between the barrel side and barrel hollow diminished with age but were still visible at P70. The changes in synaptic zinc distribution probably reflect the process of synaptic maturation of glutamatergic terminals projecting to the SI cortex. The time course of postnatal changes in terminal zinc distribution suggests that synaptic zinc is not involved in the mechanisms of barrel formation. J. Comp. Neurol. 386:652–660, 1997. © 1997 Wiley-Liss, Inc.     

Axonal trajectories between mouse somatosensory thalamus and cortex

An in vitro brain slice preparation has been used to label fibers connecting the somatosensory thalamus and cortex of the mouse. In 400-800-micron brain slices, the pathway between the ventrobasal complex and somatosensory cortex was labeled under direct vision with horseradish peroxidase crystals (HRP), HRP-Nonidet P-40 (NP40) detergent chips, or a solution of HRP/dimethylsulfoxide. Thalamocortical and corticofugal fibers are organized into a plexiform system of bundles that appears to be fairly constant from animal to animal. Bundles of fibers projecting from the ventrobasal complex course between regularly spaced groups of thalamic neurons. Thalamocortical axons do not invariably leave the thalamus via the fiber bundle closest to the perikarya. Thus, nearest-neighbor relationships are abolished before these axons have even left the thalamus. The axon bundles traverse the thalamic reticular nucleus lateral to the complex. The axons then rotate about one another, analogous to the coiling of strands in rope about a central axis. This accounts for the well known 180 degrees rotation in the mediolateral direction between thalamic and cortical maps. Laterally, fiber bundles converge and diverge within the internal capsule so that nearest-neighbor relationships are lost. Individual thalamocortical axons do not bifurcate proximal to the subcortical white matter. After single bundles of fibers reach a point just below the subcortical white matter, their individual fibers diverge widely. Within the subcortical white matter most afferent fibers make a small dorsally concave loop prior to taking one of two possible courses: some of the fibers ascend directly into the overlying cortex usually angled towards the dorsal surface of the brain; other fibers run in the subcortical white matter for variable distances prior to ascending into cortex. Within somatosensory cortex, smooth axons branch near their terminals in layers IV and VI. Axonal terminal and branching patterns of these axons within somatosensory cortex are similar to those found in in vivo preparations. Most axons are smooth, but other axons are beaded. Some beaded axons project to layer I. Corticofugal fibers are labeled. Fibers leaving somatosensory cortex have an angle of descent opposite to the angle of ascent for afferent fibers, and are often fasciculated in the cortex and subcortical white matter. Within the subcortical white matter efferent fibers often loop in a direction opposite to that of afferent fibers. Corticofugal fibers occasionally give off a collateral corticostriatal branch within the internal capsule.(ABSTRACT TRUNCATED AT 400 WORDS)     

Development of layer-specific axonal arborizations in mouse primary somatosensory cortex

In the developing neocortex, pyramidal neurons use molecular cues to form axonal arbors selectively in the correct layers. Despite the utility of mice for molecular and genetic studies, little work has been done on the development of layer-specific axonal arborizations of pyramidal neurons in mice. We intracellularly labeled and reconstructed the axons of layer 2/3 and layer 5 pyramidal neurons in slices of primary somatosensory cortex from C57Bl6 mice on postnatal days 7-21. For all neurons studied, the development of the axonal arborizations in mice follows a pattern similar to that seen in other species; laminar specificity of the earliest axonal branches is similar to that of mature animals. At P7, pyramidal neurons are very simple, having only a main descending axon and few primary branches. Between P7 and P10, there is a large increase in the total number of axonal branches, and axons continue to increase in complexity and total length from P10 to P21. Unlike observations in ferrets, cats, and monkeys, two types of layer 2/3 pyramidal neurons are present in both mature and developing mice; cells in superficial layer 2/3 lack axonal arbors in layer 4, and cells close to the layer 4 border have substantial axonal arbors within layer 4. We also describe axonal and dendritic arborization patterns of three pyramidal cell types in layer 5. The axons of tall-tufted layer 5 pyramidal neurons arborize almost exclusively within deep layers while tall-simple, and short layer 5 pyramidal neurons also project axons to superficial layers.     

Enrichment of glutamate in zinc-containing terminals of the cat visual cortex.

The presence of glutamate and GABA was examined in zinc-containing terminals of the cat visual cortex using a post-embedding immunogold method. The surface density of immunogold-labelling was also evaluated in morphologically defined ultrastructural elements, namely terminals having round synaptic vesicles and making asymmetrical synapses (RA boutons), terminals with flat vesicles and symmetrical synapses (FS) and glial cell processes. Glutamate immunoreactivity was highest in RA terminals and in zinc-containing boutons. It was lower in FS terminals and lowest in glial cell processes. GABA immunoreactivity was highest in FS terminals and low in all other ultrastructural elements analysed, including zinc-containing terminals. Therefore, zinc-containing terminals show an enrichment of glutamate and they are likely to use this amino acid as their neurotransmitter. Moreover, the fact that many RA terminals that are negative for zinc show an enrichment of immunoreactive glutamate suggests that zinc-containing fibres represent a subpopulation of the glutamate axonal network.     

Abeta-induced vascular oxidative stress and attenuation of functional hyperemia in mouse somatosensory cortex.

We investigated the role of vascular oxidative stress in the mechanisms of the impairment in cerebrovascular regulation produced by the amyloid-beta peptide (Abeta). In particular, we sought to provide evidence of vascular oxidative stress in mice overexpressing the amyloid precursor protein (APP) and to determine whether the Abeta-induced attenuation in functional hyperemia is mediated by free radical overproduction. Oxidative/nitrosative stress was assessed by 3-nitrotyrosine immunoreactivity, while free radical production was determined in cerebral microvessels by hydroethidine microfluorography. To study functional hyperemia the somatosensory cortex was activated by whisker stimulation while local blood flow was monitored by laser-Doppler flowmetry. It was found that APP mice show signs of oxidative/nitrosative stress in pial and intracerebral blood vessels well before they develop oxidative stress in neurons and glia or amyloid plaques. Treatment of cerebral microvessels isolated from wild-type mice with Abeta (1 microM) increased free radical production as assessed by the hydroethidine technique. The Abeta-induced attenuation of the increase in somatosensory cortex blood flow produced by whisker stimulation was prevented by treatment with the free radical scavengers MnTBAP or tiron. These data provide evidence that in APP mice vascular oxidative stress precedes the development of parenchymal oxidative stress, and that Abeta-produced vascular reactive oxygen species are involved in the attendant attenuation in functional hyperemia. Thus, vascular oxidative stress is an early event in the course of the brain dysfunction produced by APP overexpression and Abeta, and, as such, could be the target of early therapeutic interventions based on antioxidants.     

Development of the barrels and barrel field in the somatosensory cortex of the mouse

Barrels of the PMBSF of the mouse somatosensory cortex become apparent in Nissl-stained tangential sections simultaneously, on the fourth postnatal day. At this time they are miniatures of those in the adult and are situated in the deepest sublamina of the trilaminar cortical plate. An early barrel appears as a patch of decreased cell density: the prospective hollow of the barrel. Septa become noticeable during the sixth postnatal day. From that period to adulthood, the relative contribution of the PMBSF to the total cortical surface area increases — an increase that goes against one's expectation: the barrel related periphery matures very early and so does the central, lateral region of the cortex. Barrel growth parallel to the pial surface is greater along the major axes than along the minor axes. By using the barrels to identify prospective layer IV in immature cortex, we could determine that layers V and VI attain their adult height during the sixth postnatal day — an age when prospective layers I-IV are only half their adult height. The onset of barrel formation coincides with the moment after which injury to the pertinent somatosensory periphery (the vibrissal papillae) no longer causes profound alterations in barrel morphology.     

Lectins demarcate the barrel subfield in the somatosensory cortex of the early postnatal mouse

Plant lectins were used to examine the disposition of glycosylated molecules in vibratome sections through the barrel subfield of mouse somatosensory cortex at selected times during postnatal development. The peroxidase conjugates of peanut agglutinin (PNA, specific for N-acetylgalactosamine), concanavalin A (specific for mannose), and wheat germ agglutinin (specific for N-acetylglucosamine and N-acetylneuraminic acid) were used to study lectin binding in aldehyde-fixed tissue sections of cortex. Following peroxidase cytochemistry and light microscopy, it was found that all three lectins bound in the region of the barrel subfield as early as postnatal day 3 (day of birth = postnatal day 1). The lectins bound to the prospective sides and/or septae of individual barrels in preference to the prospective hollows. This lectin demarcation of the barrel field occurred prior to the detection of this region with cresyl violet staining and was still demonstrable on postnatal day 6, when the individual barrels became discernible with cresyl violet. This suggests that the lectin binding material is present before the barrel field becomes a fully formed and organized region. A decrease in lectin affinity for binding sites in these tissue sections occurs during postnatal development (Cooper and Steindler: Soc. Neurosci. (Abstr.) 10: 43a, '84) and this study demonstrates that lectins do not delineate the barrel field of more mature animals (2-3 months old), whereas barrels can be detected with cresyl violet at this time. A preliminary electron microscope analysis of the postnatal day 6 somatosensory cortex demonstrates that the lectin PNA binds to elements of the forming neuropil and also to Golgi apparatus intermediate saccules in neuronal cells. The prospective barrel field can be detected with lectins during a critical period in development in which alterations can occur in the barrel field in response to peripheral deprivation (Jeanmonod et al: Neuroscience 6:1503-35, '81) and therefore we suggest that the glycans visualized with lectin-peroxidase conjugates denote possible candidates for molecules involved in shaping barrel structure.     

Lesions of the basal forebrain alter stimulus-evoked metabolic activity in mouse somatosensory cortex

The role that acetylcholine plays in processing sensory stimuli is beginning to be characterized; however, morphological correlates of cholinergic effects on activity patterns in sensory cortex are not available. To study this problem, unilateral neurotoxic lesions that depleted the necortex of acetylcholine were made in the basal forebrains of mice. The aim of these experiments was to study the effect of cholinergic depletion on stimulus-evoked activity in the barrel field of the mouse somatosensory cortex. One month following the lesion, 2-deoxyglucose (2DG) experiments were conducted on the lesioned and on normal mice while the animal received bilateral stimulation to the C3 whisker. The tissue was processed for acetylcholinesterase and cytochrome oxidase histochemistry and 2DG autoradiography. Evaluation of the column-like 2DG label evoked in the somatosensory cortex revealed that the activity on the lesioned side was significantly reduced in dimension and intensity from that in the normal hemisphere. On the normal side, the activated barrels averaged 641 microns in tangential width, were 76.5% above background in density, and extended from lamina I-V. On the lesioned side, the activated barrels were 485 microns in tangential width, 65.4% above background in density, and extended from lamina II-V. In other cortical regions, outside the stimulus-evoked barrel field, 2DG activity values were similar on the normal and lesioned side. Additionally, both the pattern and intensity of the cytochrome oxidase staining within the barrel field displayed no differences in either hemisphere. These studies suggest that acetylcholine plays a significant role in the processing of somatosensory information. Neurotoxic lesions that diminish cortical cholinergic innervation cause a reduction of stimulus-evoked activity levels, while underlying metabolic activity is either not affected or recovers over time.     

Hypnotic alteration of somatosensory perception

Effects of hypnotic alterations of perception on amplitude of somatosensory event-related potentials were studied in 10 highly hypnotizable subjects and 10 subjects with low hypnotizability. The highly hypnotizable individuals showed significant decreases in amplitude of the P100 and P300 waveform components during a hypnotic hallucination that blocked perception of the stimulus. When hypnosis was used to intensify attention to the stimulus, there was an increase in P100 amplitude. These findings are consistent with observations that highly hypnotizable individuals can reduce or eliminate pain by using purely cognitive methods such as hypnosis. Together with data from the visual system, these results suggest a neurophysiological basis for hypnotic sensory alteration.     

Face representation in the human secondary somatosensory cortex.

OBJECTIVE: To investigate the somatotopic organization of the facial skin area in the secondary somatosensory cortex (SII) in humans. METHODS: Somatosensory evoked magnetic fields following air-puff stimulation of 5 body sites, the foot, the lip and 3 points of the facial skin (forehead, cheek and mandibular angle point), were recorded. We focused on activities in SII following stimulation of these 5 sites and compared dipole locations among them. RESULTS: There was a clear somatotopic organization in SII with lip in the most lateral area, foot in the most medial area and face in an intermediate area close to the lip area. However, there was no significant difference of dipole localization in SII among the 3 areas of facial skin, similar to the overlapped somatotopic organization of facial skin areas in the primary somatosensory cortex in our previous study. CONCLUSIONS: The facial skin areas are considered to occupy a small area in SII with insufficient spatial separation to differentiate each area of facial skin even using magnetoencephalography which has a high spatial resolution. SIGNIFICANCE: This is the first systematic study of the activated regions in SII following stimulation of the facial skin.     

Experience-dependent reactivation of ocular dominance plasticity in the adult visual cortex

A crucial issue in neurobiology is to understand the main mechanisms restricting neural plasticity to brief windows of early postnatal life. The visual system is one of the paradigmatic models for studying experience-dependent plasticity. The closure of one eye (monocular deprivation, MD) causes a marked ocular dominance (OD) shift of neurons in the primary visual cortex only during the critical period. Here, we report that environmental enrichment (EE), a condition of increased sensory-motor stimulation, reactivates OD plasticity in the adult visual cortex, as assessed with both visual evoked potentials and single-unit recordings. This effect is accompanied by a marked increase in cerebral serotonin (5-HT) levels. Blocking 5-HT enhancement in the visual cortex of EE rats completely prevents the OD shift induced by MD. We also found that EE leads to a reduced intracortical GABAergic inhibition and an increased BDNF expression and that the modulation of these molecular factors is neutralized by cortical infusion of the 5-HT synthesis inhibitor pCPA. Our results show that EE rejuvenates the adult visual cortex and that 5-HT is a crucial factor in this process, triggering a cascade of molecular events that allow the reinstatement of neural plasticity. The non-invasive nature of EE makes this paradigm particularly eligible for clinical application.     

Ontogeny of somatostatin receptors in the rat somatosensory cortex.

The distribution and density of SRIF receptors (SRIF-R) were studied during development in the rat somatosensory cortex by in vitro autoradiography with monoiodinated [Tyr0-DTrp8]S14. In 16-day-old fetuses (E16), intense labeling was evident in the intermediate zone of the cortex while low concentrations of SRIF-R were detected in the marginal and ventricular zones. The highest density of SRIF-R was measured in the intermediate zone at E18. At this stage, labeling was also intense in the internal part of the developing cortical plate; in contrast, the concentration of binding sites associated with the marginal and ventricular zones remained relatively low. Profound modifications in the distribution of SRIF-R appeared at birth. In particular, a transient reduction of receptor density occurred in the cortical plate. During the first postnatal week, the density of receptors measured in the intermediate zone decreased gradually; conversely, high levels of SRIF-R were observed in the developing cortical layers (II to VI). At postpartum day 13 (P13), a stage which just precedes completion of cell migration in the parietal cortex, the most intensely labeled regions were layers V-VI and future layers II-III. From P13 to adulthood, the concentrations of SRIF-R decreased in all cortical layers (I to VI) and the pattern of distribution of receptors at P21 was similar to that observed in the adults.(ABSTRACT TRUNCATED AT 250 WORDS)     

Efficient neuromagnetic determination of landmarks in the somatosensory cortex.

OBJECTIVES: An efficient procedure for the magnetoencephalographic determination of functional landmarks in the somatosensory cortex has been developed. METHODS: Digits 2-5 are stimulated in randomized order using balloon diaphragms. The interval between two stimuli is 500 ms. Source locations in area 3b are derived by interpreting the field component with a mean latency of 48 ms in terms of an equivalent current dipole. RESULTS: The signal-to-noise ratio achieved in a given time for each of the 4 stimulation sites turned out to be only slightly smaller than the one obtained by stimulating a single site with an optimal interstimulus interval (about 1 s). CONCLUSIONS: Compared to a sequential investigation of the different sites, the proposed procedure allows the reduction of the overall measurement time by a factor of about 2.7.     

Spindle wave synchrony in the somatosensory cortex of the cat.

(1) Spontaneous barbiturate spindles were recorded from the primary and secondary somatosensory cortex. The recordings were concentrated to areas surrounding several reference loci. The recording sites producing the maximal response evoked by stimulation of an exposed nerve in a contralateral limb were used as reference loci. (2) Spindles recorded at various distances from the respective reference loci were cross-correlated to spindles developing simultaneously in the latter. High correlation coefficients, indicating a considerable degree of wave synchrony, were obtained between spindles in the reference locus and spindles recorded a few millimeters from this site. The correlation coefficients decreased with increasing interelectrode distance. A relatively sharp fall in the correlation coefficients was generally found 2-3 mm from the reference locus. Small amounts of sodium pentobarbital, given intravenously at intervals of 5 min, had no effect upon this pattern. (3) The change in the correlation coefficients was followed by a parallel change in the amplitude of the evoked potentials. The iso-correlation lines of spindle wave synchrony and the iso-amplitude lines of the evoked potentials had a similar distribution and extension for each particular reference locus. (4) Lateral spread of spindle waves in the cortex seems to be of minor importance, since a vertical lesion cutting the cortico-cortical fibres did not reduce the wave synchrony of the spindles recorded from either side of the lesion. (5) The majority of the spindles recorded in the close vicinity of a reference locus started simultaneously within +/- 0.1 sec. This pattern changed with increasing distance from the reference locus and 5.6 mm away only a fraction of the spindles started simultaneously. However, within the entire primary somatosensory cortex a small but significant coupling existed between onset of the spindles.