Deprivation and denervation differentially affect zinc-containing circuitries in the barrel cortex of mice

In the neocortex, a population of glutamatergic synapses contains chelatable zinc that is released upon depolarization. The present study compares the effect of chronic tactile deprivation and vibrissectomy performed at different postnatal ages on the synaptic zinc distribution in the mouse barrel cortex. We found that a chronic unilateral tactile deprivation resulted in an increase of synaptic zinc in deprived barrels. Distribution and intensity of zinc staining in non-deprived barrels resembled the control situation. The increase of zinc staining was observed if chronic deprivation started in early postnatal life or in adolescent mice but not in 70-day-old animals. This suggests that a critical period exists for plasticity of zinc containing terminals in the barrel cortex. The alteration of zinc staining was localized to not only the thalamorecipient layers IV but also layer II/III, and upper layer V. Neonatal denervation of selected vibrissal rows resulted in rearrangement of synaptic zinc distribution following cytoarchitectonic alterations in the barrel field. However, no changes in the intensity of zinc staining were observed. Vibrissectomy performed after the critical period for barrel formation did not affect either the distribution or intensity of zinc staining. It appears that the integrity of vibrissa-barrel pathway is necessary to induce activity-dependent alterations in synaptic zinc.     

Histochemical localization of synaptic zinc in the developing cat visual cortex

The terminal boutons of many neurons in the telencephalon are known to contain a vesicle-bound, chelatable pool of zinc (Zn²⁺) that can be selectively visualized with histochemical procedures. In this paper, the normal laminar, areal, and ultrastructural distribution of histochemically reactive zinc in the visual cortex of the adult cat as well as its development from birth are described. In the adult cat visual cortex, intense zinc staining was found in layers I, II, III, and V, with layer VI staining only lightly. The primary geniculostriate input zone, layer IV, was conspicuously distinguished by the relative absence of zinc. This distinct pattern was restricted only to areas 17 and 18 and differentiated them from adjacent cortical area 19 laterally and the subadjacent cingulate cortex. The earliest zinc-positive staining in visual cortical areas 17 and 18 was first apparent by postnatal day 2 (P2) and was characterized by staining of a thin layer at the bottom of the cortical plate. By P10, and continuing through P20, synaptic zinc formed a trilaminar pattern of dense staining in areas 17 and 18, which included the top of layer I, and layers III and V. The laminar pattern of synaptic zinc in visual cortex appeared mature by P30, except that the distribution of zinc in layer IV was not uniform. This was most apparent around P50 in tangential sections through layer IV from opened and flattened cortex, where columnar patches of increased zinc staining were apparent in area 17. These columns were approximately 400 μm in diameter, with a centre-to-centre spacing of approximately 900 μm. The distribution of synaptic zinc apparently reflects the process of synaptic maturity of the cat visual cortex and appears to demarcate a particular form of columnar organization in visual cortex. © 1993 Wiley-Liss, Inc.     

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.     

Developmental patterns of cadherin expression and localization in relation to compartmentalized thalamocortical terminations in rat barrel cortex

The wiring of synaptic circuitry during development is remarkably precise, but the molecular interactions that enable such precision remain largely to be defined. Cadherins are cell adhesion molecules hypothesized to play roles in axon growth and synaptic targeting during development. We previously showed that N-cadherin localizes to ventrobasal (VB) thalamocortical synapses in rat somatosensory (barrel) cortex during formation of the whisker-map in layer IV (Huntley and Benson [1999] J. Comp. Neurol. 407:453-471). Such specific association of N-cadherin with one identified afferent pathway raises the prediction that other cadherins are expressed in barrel cortex and that these are, in some combination, also differentially associated with distinct inputs. Here, we first show that N-cadherin and three other classic cadherins (cadherin-6, -8, and -10) are expressed contemporaneously in barrel cortex with relative levels of postnatal expression that are highest during the first 2 weeks, when afferent and intrinsic circuitries are forming and synaptogenesis is maximal. Each displayed distinct, but partly overlapping laminar patterns of expression that changed over time. Cadherin-8 probe hybridization formed a particularly striking pattern of intermittent, columnar patches extending from layer V through layer III, which was first detectable at approximately postnatal day 3. The patches were centered precisely over regions of dysgranular layer IV and, in the whisker barrel field, over barrel septa. This pattern is similar to that formed by the terminal distribution of thalamocortical afferents arising from the posterior nucleus (POm), suggesting cadherin-8 association with the POm thalamocortical synaptic circuit. Consistent with this, cadherin-8 mRNAs were enriched in the POm nucleus, and cadherin-8 immunolabeling in layer IV was enriched in barrel septa and codistributed with labeled POm thalamocortical synaptic-like puncta. The striking molecular parcellation of at least two different cadherins to the two, converging thalamic pathways that terminated in non-overlapping barrel center and septal compartments in layer IV strongly suggested that cadherins provide requisite molecular recognition and targeting that enable precise construction of thalamocortical and other synaptic circuitry.     

Development of laminar distributions of kainate receptors in the somatosensory cortex of mice

Kainate receptors were present at birth in the murine somatosensory cortex as revealed by quantitative in vitro autoradiography. During the first five postnatal days [³H]kainate binding rapidly increased and the maximum density in layer IV was reached at P12. The adult laminar pattern of receptor binding distribution was established by the third postnatal week with the heaviest labeling of infragranular layers. The sharp increase of kainate receptor during the first postnatal week coincides with the critical period for cytoarchitectonic plasticity of the barrels and establishment of functional thalamo-cortical connections in the barrel field.     

gamma-Aminobutyric acid (GABA) immunoreactivity in mouse and rat first somatosensory (SI) cortex: description and comparison

The location and morphological characteristics of gamma-aminobutyric acid (GABA)-immunopositive cells and their processes were studied in rat and mouse first somatosensory (SI) cortex (including 'barrels') in layer IV, and layers above (I-III), and below (V and VI). In coronal sections of SI cortex of both species GABA-immunopositive cells and punctate profiles were found in each of layers I-VI. The cells were of various sizes; the largest, located in layers III and V of each species, resemble the large basket cells seen in Golgi-impregnated material. Most of the immunopositive cells were multipolar and circular or ellipsoidal in shape, but occasionally bipolar cells with fusiform perikarya were also seen. In coronal sections, immunopositive cells did not form a characteristic pattern. GABA-immunopositive cells were observed to be most numerous in the supragranular layers whereas GABA-positive punctate profiles were more numerous in layer IV. In tangential sections from layer IV of SI cortex of both species, GABA-immunopositive cells, processes and punctate profiles were visible throughout the entire barrel field. The pattern of distribution of immunopositive cells was similar (a) in two different morphological groups--i.e. the posteromedial barrel subfield (PMBSF) and the anterolateral barrel subfield (ALBSF) in rat barrel field, and (b) in PMBSF barrels of both rat and mouse (excluding differences due to structural dissimilarities between rat and mouse barrels). GABA-immunopositive neurons were grouped mainly in the barrel side and septum and were visible frequently in small clusters. In barrels of both species GABA-immunopositive cells were of a variety of sizes and ranged in shape from ellipsoidal to circular.     

Expression of two forms of glutamic acid decarboxylase (GAD67 and GAD65) during postnatal development of rat somatosensory barrel cortex

The postnatal development of glutamic acid decarboxylase (GAD; GAD67 and GAD65) expression was studied in the rat somatosensory cortex. Delineation of barrels in layer IV by GAD67 immunoreactivity occurred between postnatal days P3 and P6 and remained evident into adulthood. At birth, a band of GAD67-positive elements was already present in superficial layer V. This band was prominent until P6 and gradually disappeared after P9. In parallel, there was a gradual appearance of GAD67-immunoreactive cells neuropil and puncta, which began in layer VI/subplate at P1 and achieved the adult laminar pattern by about P13. This later GAD67 immunoreactivity was responsible for the demarcation of barrels in layer IV. Development of GAD65 immunoreactivity was delayed relative to GAD67. GAD65 immunoreactivity, which was in little evidence before P6, increased markedly in density and in delineation of cell bodies over the next several weeks. During this prolonged developmental process, GAD65 first showed a negative image of the barrels compared with the septae and the surrounding cortex. Subsequently, there was a filling in of the barrels resulting in rather uniform GAD65 immunoreactivity across the barrel field and surrounding cortex. These results suggest that the development of the γ-aminobutyric acid (GABA) synthetic system in the barrel cortex involves several processes: the disappearance of a precocious GAD67 system in layer V, the temporally overlapping maturation of the mature GAD67 system in an inside-outside manner, and the delayed and prolonged development of the GAD65 system. J. Comp. Neurol. 402:62–74, 1998. © 1998 Wiley-Liss, Inc.     

Ontogeny of non-NMDA glutamate receptors in rat barrel field cortex: I. metabotropic receptors

The ontogeny of metabotropic excitatory amino acid receptors (mGluRs) in rat barrel field cortex was characterized by using receptor autoradiography and immunocytochemistry to test the hypothesis that changes in mGluR expression coincide with the emergence of somatotopic patterns in this region. On postnatal days 1 (P1) and 3, [³H]glutamate binding to mGluRs was not distributed in a somatotopic pattern. By P5, mGluRs exhibited a whisker-related pattern, with higher densities of mGluRs in barrel centers than in surrounding cortex. Between P5 and P14 and at P60, the overall binding density remained higher in barrels than in surrounding cortex. At P60, a somatotopic pattern of binding was not apparent. The majority of mGluR sites in the barrel field were blocked by the metabotropic agonist trans-1-aminocyclopentane-1,3-dicarboxylic acid but were not significantly displaced by quisqualate. Immunocytochemical studies of phosphoinositide-linked mGluRs, mGluR5 and mGluR1α, showed that the developmental expression of mGluR5 mirrored that of the pattern of autoradiographically labeled mGluRs. The immature barrel field (ages P5–P14) was enriched in mGluR5, with greater concentrations of mGluR5 immunoreactivity in barrels than in surrounding cortex. Within barrel centers, mGluR5 was localized within the neuropil, on the surfaces of cell bodies and dendrites in layer IV. A somatotopic pattern of mGluR5 immunoreactivity persisted into adulthood, although the pattern was less pronounced after P14. In contrast, mGluR1α was never localized in a somatotopic pattern in barrel field cortex. We conclude from the developmental localization of mGluRs that the spatiotemporal regulated expression of these receptors may influence barrel maturation and plasticity. J. Comp. Neurol. 386:16–28, 1997. © 1997 Wiley-Liss, Inc.     

Postnatal development of zinc-rich terminal fields in the brain of the rat

The appearance and distribution of zinc-rich terminal fields in the rat forebrain was analyzed at 12 stages of postnatal development using the selenium method. Zinc stain was detected in neonates in piriform, cingulate, and motor cortices, septal area, and hippocampal formation. In the neocortex, a laminar pattern appeared progressively following an inside-out gradient: layer VI at postnatal day 0 (P0), layer V at P1, layers Va and Vb at P5, layer II-III at P9, and layer IV at P12. In the hippocampal formation the layered pattern in the dentate molecular layer appeared at P1-P3, and in the hilus and mossy fibers the stain was observed at P5. Patches in the caudate-putamen were sharply delimited at P1-P3. At these ages, staining was observed in the amygdaloid complex. In the thalamic and hypothalamic nuclei, stain appeared at P5-P7. Thus, a general increase in vesicular zinc over different telencephalic areas was determined until P15-P21, which was followed by a slight decrease thereafter (at P41). The increased stain in zinc-rich terminal fields is consistent with the development of telencephalic circuits. The rise in zinc might be relevant for the establishment and maturation of these circuits. On the other hand, the decrease in staining for zinc at later stages might be due to methodological problems but it might also reflect pruning of supernumerary connections and programmed cell death affecting zinc-rich circuits.     

Spatiotemporal distribution of proteoglycans in the developing rat's barrel field and the effects of early deafferentation

The isolectin Vicia villosa B(4) (VV) selectively recognizes N-acetyl-galactosamine-terminal glycoconjugates that form perineuronal nets (PNNs) around a subset of neurons in the cerebral cortex. PNNs are thought to participate in the guidance of incoming thalamic axons and in the posterior stabilization and maintenance of synaptic contacts. Here we examine the spatial and temporal distribution of biotinylated VV in tangential sections through layer IV of the posteromedial barrel subfield in the primary somatosensory cortex (PMBSF) of rats ranging from postnatal day (P)3 to P60, which underwent unilateral deafferentation of whiskers at birth. In the afferented hemisphere, labeling first appears at P5, with a diffuse distribution, probably associated with neuropil, inside PMBSF barrels. VV distribution remains diffuse during the following week, and declines around P17. From P24 onward, however, proteoglycans form PNNs around cell bodies preferentially localized in septal regions of the PMBSF. In the contralateral, deafferented PMBSF the diffuse labeling also appears on P5, but first develops into elongated, homogeneous stripes, which disappear after P24, leaving only scattered cell bodies along layer IV. Our results indicate that proteoglycans appear simultaneous to barrel formation in the developing rat while segregation of PNNs to septal cells might be driven by afferent activity.     

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.     

Experience-dependent alteration of zinc-containing circuits in somatosensory cortex of the mouse.

Histochemical staining was used to localize zinc-sequestering terminals in somatosensory barrel cortex of normal mice and mice subjected to tactile deprivation by simple whisker trimming from birth. In normal mice, density of synaptic zinc was highest in laminae I, II and V, intermediate in laminae III and VI, and lowest in lamina IV barrel hollows. Whisker trimming from birth led to increased density of synaptic zinc specifically within deprived barrel hollows.     

Developmental distribution of calretinin in mouse barrel cortex

We describe the postnatal development of calretinin expression in the mouse barrel cortex by immunohistochemistry. A densely staining neuropil and numerous cell bodies appeared throughout layer V, but only within barrel septa of layer IV, at postnatal day 4. This staining pattern became most robust at postnatal day 8. Thereafter, calretinin expression became reduced until the third postnatal week when it attained its mature levels, and the barrel-specific staining was no longer apparent.     

Septal columns in rodent barrel cortex: functional circuits for modulating whisking behavior

In rodents, each mystacial whisker is represented in the granular layer of primary somatosensory (SI) cortex by a compact cluster of cells known as a barrel, and barrels are separated from each other by domains that are called septa. Vertical columns of neurons aligned with each barrel act as a functional assembly to process information from a "principal" whisker, but a functional role has not been identified for vertical columns of neurons that are aligned with the septa. To determine whether these septal columns provide the main source of projections to primary motor (MI) cortex, we placed retrograde tracers in MI cortex and analyzed the location of the retrogradely labeled neurons with respect to the septal and barrel compartments of SI barrel cortex. In cases in which SI barrel cortex was sectioned tangentially, retrogradely labeled neurons in the extragranular layers of SI were plotted and superimposed onto reconstructions of the layer IV barrel field. In each of these cases, most labeled neurons were located above or below the septal regions of layer IV. When SI barrel cortex was sectioned coronally, we observed multiple columns of labeled SI neurons that were vertically aligned with the septal zones of layer IV. These results indicate that columns of neurons that are vertically aligned with the septa, or septal columns, are functionally linked by virtue of their projections to MI cortex. We hypothesize that these septal columns represent an interconnected and functionally distinct circuit that transmits information to MI and other brain regions involved in motor control.     

Sensitive period for lesion-induced reorganization of intracortical projections within the vibrissae representation of rat's primary somatosensory cortex

Previous experiments from this laboratory demonstrated that intracortical connections in lamina IV of the rat primary somatosensory cortex (SI) are most dense outside the patches of cytochrome oxidase (CO) staining that correspond to the mystacial vibrissae. This pattern of intracortical connections becomes apparent on postnatal day 4 (P-4), at least 2 days after the appearance of the vibrissae-related pattern of thalamocortical afferents. Transection of the infraorbital nerve (ION) on the day of birth (P-0) disrupts both the CO and intracortical projection patterns. This series of experiments was undertaken to determine whether the patterning of either thalamocortical afferents or intracortical projections defines the end of the period over which peripheral damage can alter intracortical projections in lamina IV of SI. The infraorbital nerve (ION) was transected in different cohorts of rats on P-1 through P-5, and animals were allowed to survive ≥45 days, at which time biotinylated dextran amine (BDA) injections were made into the SI. After 7 days, animals were killed, and alternate cortical sections were processed for the demonstration of BDA or CO. Transection of the ION on P-1 or P-2 altered the patterning of both CO and intracortical connections in the SI. In contrast, cutting the ION on P-3 left the pattern of CO densities in the SI intact, but significantly altered the patterning of intracortical connections. Transection of the nerve on P-5 resulted in qualitatively and quantitatively normal patterns of both CO densities and BDA-labelled intracortical projections. These results indicate that the establishment of a stable barrel pattern in layer IV of the SI is not sufficient for normal adult patterning of intracortical projections in this lamina. However, once the mature pattern of intracortical projections in layer IV is established, ION lesions can no longer alter it. J. Comp. Neurol. 389:185–192, 1997. © 1997 Wiley-Liss, Inc.     

Developmental expression of GABAA receptor subunit and GAD genes in mouse somatosensory barrel cortex

In situ hybridization histochemistry with radioactive cRNA probes was used to study patterns of gene expression for α1, α2, α4, α5, β1, β2, and γ2 subunit mRNAs of type A gamma aminobutyric acid (GABA_A) receptors and for 67-kDa glutamic acid decarboxylase (GAD₆₇) mRNA in mouse barrel cortex during the period (postnatal days 1-12; P1-P12) when thalamocortical innervation of layer IV barrels is occurring. The α1, β2, and γ2 subunit mRNAs increased substantially with age, especially in layers V and VI, and throughout the period studied, invariably had the same laminar-specific patterns of expression. All three mRNAs were highly expressed in the dense cortical plate at P1. In layer IV after differentiation of barrels, they were expressed in cells of both barrel walls and hollows but especially in the walls. The α2, α4, α5, and β1 subunit mRNAs were expressed at lower levels and had different laminar patterns of distribution; α2 and α4 showed switches between layers over time; α5 was invariably associated with the subplate or its derivative, β1 with layer IV. Levels of α2 mRNA did not change over time; α4 and β1 mRNAs increased and α5 decreased. GAD₆₇ mRNA was highest in layer I at P1 and progressively increased in other layers. These results suggest that postnatal development of GABA_A receptors is mainly directed at the production of receptors assembled from α1, β2, and γ2 subunits, with β1 contributing in layer IV. Other subunits may be associated with receptors involved in trophic actions of GABA during development and may give GABA_A receptor-mediated responses in the developing cortex their particular physiological profile. J. Comp. Neurol. 383:199-219, 1997. © 1997 Wiley-Liss, Inc.     

Altered zincergic innervation of the developing primary somatosensory cortex in monoamine oxidase-A knockout mice

Genetic inactivation of monoamine oxidase-A (MAO-A) significantly elevates levels of serotonin (5-HT) during early development and causes a disruption in the compartmented organization of thalamocortical axon terminals in layer 4 of the somatosensory cortex. In order to determine whether corticocortical innervation of the primary somatosensory cortex is also affected by this mutation, we examined the distribution of zinc-containing axon terminals (terminals known to originate from within the cortex) in the developing somatosensory cortex of MAO-A knockout mice, at postnatal days (PD) 3, 5, 6, 8, 10, 12, 15, 28, and 60. In layer 4 of wild-type mice, histochemical staining for zinc respected barrel-specific compartments at all ages beyond PD 5. By contrast, zinc staining in MAO-A knockout mice did not exhibit signs of barrel compartmentation at any age. Across cortical layers, substantial developmental changes in the distribution of zinc-containing terminals were observed in wild-type mice up until PD 12, at which time the mature lamina-specific pattern of zinc staining was achieved. Similar changes were observed in the somatosensory cortex of MAO-A knockout mice, except that its developmental time course was significantly compressed, with zincergic innervation achieving a mature appearance by PD 8. These results provide evidence that an excess of monoamines, most likely 5-HT, dramatically perturbs the columnar organization of intracortical zincergic afferents in layer 4 and significantly accelerates the appearance of a mature laminar pattern of zinc-containing corticocortical terminals.     

Primary auditory cortex in the rat: transient expression of acetylcholinesterase activity in developing geniculocortical projections

A characteristic pattern of acetylcholinesterase (AChE) activity is expressed transiently in primary auditory cortex (cortical area 41) of developing laboratory rats during early postnatal life. This AChE activity occurs as a dense plexus in cortical layer IV and the deep part of layer III. This transient band of AChE activity is first detected by histochemical techniques on postnatal day (P) 3, reaches peak intensity at approximately P8-10, and declines to form the adult pattern by P23. The ventral nucleus of the medial geniculate body of the thalamus also displays prominent, and transient, staining for AChE. This intense staining for AChE, found within neuronal somata and neuropil, is detected at the time of birth, reaches peak intensity around P8, and declines to adult levels by P16. The areal and laminar patterns of the transient band of AChE activity in temporal cortex correspond to the patterns of anterograde transneuronal labeling of geniculocortical terminals following injection of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) into the inferior colliculus. Placement of lesions that include the medial geniculate nucleus or the geniculocortical axons results in a marked decrease in AChE staining in thalamorecipient layers of auditory cortex. Placement of lesions that include the medial globus pallidus reduce AChE staining of some axons in temporal cortex of developing rats, but the dense band of AChE in layers III and IV remains. Placement of lesions in the inferior colliculus in newborn animals results in marked decrease in AChE staining in cells of the ipsilateral ventral medial geniculate nucleus and in ipsilateral auditory cortex of developing pups. These data indicate that transiently expressed AChE activity is characteristic of geniculocortical neurons, including their somata in the medial geniculate body and their terminal axons in primary auditory cortex. This AChE activity is expressed early in postnatal development, probably during the time when thalamocortical axons are proliferating in cortical layer IV and forming synaptic contacts with cortical neurons.     

Distribution of GABAergic Elements Postsynaptic to Ventroposteromedial Thalamic Projections in Layer IV of Rat Barrel Cortex

The spatial synaptic pattern formed by boutons, originating in the ventroposteromedial thalamic nucleus, with GABAergic neurons in the rat barrel cortex was mapped. The aim was to shed light on the structural basis by which inhibitory circuits may be activated at the first stage of cortical information processing. The thalamic afferent projection was labelled by anterograde transport of Phaseolus vulgaris leucoagglutinin (PHA-L), whereas the GABAergic targets in layer IV of the rat barrel cortex were visualized by postembedding GABA immunogold-labelling or by pre-embedding parvalbumin immunocytochemistry. In the first set of experiments, we mapped barrels, contained in single ultrathin sections, by means of a computer-controlled electron microscope stage in their entire layer IV representation. From a total of 1199 asymmetric PHA-L-labelled synapses, only 98 were on GABAergic elements, mainly on dendritic shafts. This corresponded to 8.2% of all synapses counted. These synapses on GABAergic targets were essentially homogeneously distributed without a reliable relationship to barrel subdivisions, i.e. hollow versus wall; or layer IVa versus layer IVb. In the second part of the study, we demonstrated that parvalbumin-containing neurons represent the major GABAergic cell type targetted by thalamic afferents in layer IV of the barrel cortex, since all parvalbumin-positive cells investigated received multiple synaptic contacts (up to eight synapses per neuron) from the ventroposteromedial thalamic nucleus. These results imply that interneurons responsible for perisomatic inhibition (basket and chandelier cells known to contain parvalbumin) are likely to be strongly excited by thalamic afferents, despite the relatively low proportion of thalamic synapses on GABAergic elements compared to spines of principal cells, and participate in the early stages of cortical sensory information processing.     

Long-term potentiation in the neonatal rat barrel cortex in vivo

Long-term potentiation (LTP) is important for the activity-dependent formation of early cortical circuits. In the neonatal rodent barrel cortex, LTP has been studied only in vitro. We combined voltage-sensitive dye imaging with extracellular multielectrode recordings to study whisker stimulation-induced LTP in the whisker-to-barrel cortex pathway of the neonatal rat barrel cortex in vivo. Single whisker stimulation at 2 Hz for 10 min induced an age-dependent expression of LTP in postnatal day (P) 0 to P14 rats, with the strongest expression of LTP at P3-P5. The magnitude of LTP was largest in the activated barrel-related column, smaller in the surrounding septal region, and no LTP could be observed in the neighboring barrel. Current source density analyses revealed an LTP-associated increase of synaptic current sinks in layer IV/lower layer II/III at P3-P5 and in the cortical plate/upper layer V at P0-P1. Our study demonstrates for the first time an age-dependent and spatially confined LTP in the barrel cortex of the newborn rat in vivo.