Somata Of Basket Cells Research Articles

Axons of cortical basket cells originating from dendrites develop higher local complexity than axons emerging from basket cell somata.

Neuronal differentiation is regulated by neuronal activity. Here, we analyzed dendritic and axonal growth of Basket cells (BCs) and non-Basket cells (non-BCs) using sparse transfection of channelrhodopsin-YFP and repetitive optogenetic stimulation in slice cultures of rat visual cortex. Neocortical interneurons often display axon-carrying dendrites (AcDs). We found that the AcDs of BCs and non-BCs were, on average, the most complex dendrites. Further, the AcD configuration had an influence on BC axonal development. Axons originating from an AcD formed denser arborizations with more terminal endings within the dendritic field of the parent cell. Intriguingly, this occurred already in unstimulated BCs, and complexity was not increased further by optogenetic stimulation. However, optogenetic stimulation exerted a growth-promoting effect on axons emerging from BC somata. The axons of non-BCs neither responded to the AcD configuration nor to the optogenetic stimulation. The results suggest that the formation of locally dense BC plexuses is regulated by spontaneous activity. Moreover, in the AcD configuration, the AcD and the axon it carries mutually support each other's growth.

Hyperpolarization-activated currents in presynaptic terminals of mouse cerebellar basket cells.

Using patch-clamp techniques, a hyperpolarization-activated current (I(h)) was recorded from synaptic terminals of mouse cerebellar basket cells. Ih was blocked quickly and reversibly by 2 mM Cs(+), and subtraction revealed a rapidly activating and deactivating I(h) current. Similar gating and block of presynaptic I(h) were also seen with the more selective inhibitor ZD 7288 (10 microM). The time constant of activation (tau (a))of presynaptic I(h) current became faster with membrane hyperpolarization, being approximately 74 ms at -130 mV, changing e-fold for a 33 mV change in membrane potential. Whole-cell recordings from basket cell somata also revealed an I(h) current, which was similarly sensitive to block by ZD 7288. Inhibition of I(h) by 10 microM ZD 7288 reduced the frequency ( approximately 34 %) and amplitude ( approximately 26 %) of spontaneous IPSCs (sIPSCs) recorded in Purkinje cells, one of the principal synaptic targets of basket neurones. This is the first report of an I(h) current in mammalian inhibitory presynaptic terminals, which may be an important target for neuromodulation in the cerebellum. Comparing the biophysical properties and distribution of cloned hyperpolarization-activated cation channels, we also suggest a molecular candidate underlying I(h) at these synapses.

Ca(2+)-permeable AMPA and NMDA receptor channels in basket cells of rat hippocampal dentate gyrus.

1. Glutamate receptor (GluR) channels were studied in basket cells in the dentate gyrus of rat hippocampal slices. Basket cells were identified by their location, dendritic morphology and high frequency of action potentials generated during sustained current injection. 2. Dual-component currents were activated by fast application of glutamate to outside-out membrane patches isolated from basket cell somata (10 microM glycine, no external Mg2+). The fast component was selectively blocked by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), the slow component by D-2-amino-5-phosphonopentanoic acid (D-AP5). This suggests that the two components were mediated by alpha-amino-3- hydroxy-5-methyl-4-isoxazolepropionate receptor (AMPAR)/kainate receptor and N-methyl-D-aspartate receptor (NMDAR) channels, respectively. The mean ratio of the peak current of the NMDAR component to that of the AMPAR/kainate receptor component was 0.22 (1 ms pulses of 10 mM glutamate). 3. The AMPAR/kainate receptor component, which was studied in isolation in the presence of D-AP5, was identified as AMPAR mediated on the basis of the preferential activation by AMPA as compared with kainate, the weak desensitization of kainate-activated currents, the cross-desensitization between AMPA and kainate, and the reduction of desensitization by cyclothiazide. 4. Deactivation of basket cell AMPARs following 1 ms pulses of glutamate occurred with a time constant (tau) of 1.2 +/- 0.1 ms (mean +/- S.E.M.). During 100 ms glutamate pulses AMPARs desensitized with a tau of 3.7 +/- 0.2ms. 5. The peak current-voltage (I-V) relation of AMPAR-mediated currents in Na(+)-rich extracellular solution showed a reversal potential of -4.0 +/- 2.6 mV and was characterized by a a doubly rectifying shape. The conductance of single AMPAR channels was estimated as 22.6 +/- 1.6 pS using non-stationary fluctuation analysis. AMPARs expressed in hippocampal basket cells were highly Ca2+ permeable (PCa/PK = 1.79). 6. NMDARs in hippocampal basket cells were studied in isolation in the presence of CNQX. Deactivation of NMDARs activated by glutamate pulses occurred bi-exponentially with mean tau values of 266 +/- 23 ms (76%) and 2620 +/- 383 ms (24%). 7. The peak I-V relation of the NMDAR-mediated component in Na(+)-rich extracellular solution showed a reversal potential of 1.5 +/- 0.6 mV and a region of negative slope at negative membrane potentials in the presence of external Mg2+, due to voltage-dependent block by these ions. The conductance of single NMDAR channels in the main open state was 50.2 +/- 1.8 pS.(ABSTRACT TRUNCATED AT 400 WORDS)

Laminar distribution and neuronal targets of GABAergic axon terminals in cat primary auditory cortex (AI)

The form, density, and neuronal targets of presumptive axon terminals (puncta) that were immunoreactive for gamma-aminobutyric acid (GABA) or its synthesizing enzyme, glutamic acid decarboxylase (GAD), were studied in cat primary auditory cortex (AI) in the light microscope. High-resolution, plastic-embedded material and frozen sections were used. The chief results were: 1) There was a three-tiered numerical distribution of puncta, with the highest density in layer Ia, an intermediate number in layers Ib-IVb, and the lowest concentration in layers V and VI, respectively. 2) Each layer had a particular arrangement: layer I puncta were fine and granular (less than 1 micron in diameter), endings in layers II-IV were coarser and more globular (larger than 1 micron), and layer V and VI puncta were mixed in size and predominantly small. 3) The form and density of puncta in every layer were distinctive. 4) Immunonegative neurons received, in general, many more axosomatic puncta than immunopositive cells, with the exception of the large multipolar (presumptive basket) cells, which invariably had many puncta in layers II-VI. 5) The number of puncta on the perikarya of GABAergic neurons was sometimes related to the number of puncta in the layer, and in other instances it was independent of the layer. Thus, while layer V had a proportion of GABAergic neurons similar to layer IV, it had only a fraction of the number of puncta; perhaps the intrinsic projections of supragranular GABAergic cells are directed toward layer IV, as those of infragranular GABAergic neurons may be. Since puncta are believed to be the light microscopic correlate of synaptic terminals, they can suggest how inhibitory circuits are organized. Even within an area, the laminar puncta patterns may reflect different inhibitory arrangements. Thus, in layer I the fine, granular endings could contact preferentially the distal dendrites of pyramidal cells in deeper layers. The remoteness of such terminals from the spike initiation zone contrasts with the many puncta on all pyramidal cell perikarya and the large globular endings on basket cell somata. Basket cells might receive feed-forward disinhibition, pyramidal cells feed-forward inhibition, and GABAergic non-basket cells would be the target of only sparse inhibitory axosomatic input. Such arrangements imply that the actions of GABA on AI neurons are neither singular nor simple and that the architectonic locus, laminar position, and morphological identity of a particular neuron must be integrated for a more refined view of its role in cortical circuitry.

An analysis of GABAergic afferents to basket cell bodies in the cat's cerebellum

An antibody to glutamic acid decar☐ylase (GAD) was used to identify GABAergic terminals around the somata of basket cells in the cat's cerebellar cortex. Two sources for GAD immunoreactive terminals were identified based on size and cytological characteristics including the recurrent collaterals derived from Purkinje cells and the axons of stellate cells. The majority of the unlabeled terminals likely arise from parallel fibers. The present analysis indicates that GAD-positive terminals form 13% of the synaptic contacts on basket cells. The remaining 87% are unlabeled. Thus, GAD-positive terminals represent a small proportion of the synaptic input to basket cell bodies as compared to afferent endings that likely mediate excitation.

Cytology and organization of rat cerebellar organ cultures

Roller tube cultures of parasagittal cerebellar slices were taken from young rats aged 9–11 days, and maintained in vitro for 1–2 weeks. Morphological aspects of cell types and synaptic relationships in such organ cultures were examined at light and electron microscopic levels. Some neurons were marked by intracellular injections of horseradish peroxidase for subsequent identification of their connection patterns. Cytoarchitecture of the cerebellar cortex was largely preserved in the organ cultures. Dendritic trees of Purkinje cells exhibited isoplanar organizations that often resembled their orientation at the time of explanation. Other cerebellar neurons, namely granule cells, Golgi cells, basket cells, stellate cells, all differentiated within the organ cultures. In addition, some neurons of the deep cerebellar nuclei remained viable during the period of culture. Mossy fibers most probably of cerebellar nuclear origin were found terminating on the dendrites of granule cells and Golgi cells. Quite unexpected were certain types of direct synapses of afferent fibers on short necked spines arising from Purkije cell smooth dendrites and somata. Such terminals resembled climbing fibers. They were most likely modified mossy fiber afferents, since the organ cultures did not include neurons of the inferior olive which are well spearated from the cerebellar mass at postnatal stages. These “ascending” mossy fibers presumably occupied postsynaptic surfaces that were either vacated by deafferentation or induced by the afferent fibers themselves. Intracellularly labeled Purkinje cells had widely distributed axonal collateral branches. Labeled axons were distributed within the Purkinje cell layer. Several recurrent Purkinje cell axon collaterals stained with reaction products of horseradish peroxidase tracer were followed at the ultrastructural level. In one case, labeled terminals were examined in an area of approximately 2 mm 2. Terminals of Purkinje cell collaterals formed symmetric synapses with somata of basket cells and dendrites of Golgi cells, but not Purkinje cell somata. Some large boutons of serially traced Purkinje cell axon collaterals formed asymmetric contacts with profiles interpreted as Golgi cell dendrites. In contrast to the apparent axonal sprouting in cerebellar organ cultures, maturation of dendritic processes remained static. Astroglia cells of diverse shapes were observed following immunocytdchemical staining with antisera to glia filament proteins. The distribution patterns of immunoreactive astrocytes changed dramatically in cerebellar slice cultures maintained for 3–6 weeks in vitro.

Thyroid hormone modulates the expression of a neurofilament antigen in the cerebellar cortex: premature induction and overexpression by basket cells in hyperthyroidism and a critical period for the correction of hypothyroidism

Neurofilament expression by basket cells of the cerebellar cortex is suppressed in hypothyrodism. By using a monoclonal antibody (mabN210) that selectively recognizes an epitope associated with the 210-kDa neurofilament subunit, we have explored the relationship between thyroid hormone levels and basket cell maturation. In animals rendered hypothyroid by inclusion of propylthiouracil in the maternal drinking water from embryo age E17, there is a complete absence of mabN210 immunoreactivity in the basket cell axons, while the other immunoreactive axons in the cerebellar cortex, primarily Purkinje cell axons and mossy fibers, are apparently unaffected. This deficit can be corrected by treatment with thyroid hormone but there seems to be a critical period for full recovery, for animals treated from birth recover normally whereas there is a gradual diminution in the efficacy of treatment the later it begins. Thyroid hormone therapy begun after postnatal day 30 (P30) leads only to very minor recovery. By contrast, animals on a hyperthyroid regime show premature mabN210-antigen induction in the basket cells and supranormal levels of expression at P25, despite the severe reduction in the number of basket cell somata. This suggests either abnormal compensatory sprouting of axon collaterals by the remaining basket cells or the occurrence, during normal cerebellar corticogenesis, of competition between basket cell axons for a limited number of Purkinje cell targets followed by the elimination of the excess collaterals.

Five types of basket cell in the hippocampal dentate gyrus: a combined Golgi and electron microscopic study.

Five types of basket cell in the hippocampal denate gyrus of rats were analysed with a combined Golgi and electron microscopic method. Light microscopic observations show that the large somata of these different cell types are located either in the granule cell layer or within 30-50 micron of this layer. The somata of basket cells are pyramidal, horizontal, fusiform or multipolar. Dendrites of basket cells are aspinous or sparsely spinous and are found in all layers of the dentate gyrus. Their axons form an extensive plexus in the granule cell and lower molecular layers. Electron microscopic preparations of Golgi-impregnated, gold-toned basket cells revealed gold-labelled neurons with distinct ultrastructural features. All somata of basket cells displayed an extensive perikaryal cytoplasm with large Nissl bodies and nuclei with infoldings, euchromatin, intranuclear rods and sheets, and large nucleoli. The aspinous dendrites as well as the somata had a mixture of asymmetric and symmetric synapses on their surfaces. Basket cell dendrites located in the hilus were contacted by numerous terminals with characteristics of mossy fibres derived from granule cells. Some of these terminals were identified positively in preparations that also contained impregnated granule cells. The axons of basket cells formed exclusively symmetric synapses. The most common postsynaptic structures to these terminals were the somata and dendrites of granule cells. Dendritic spines were rarely contacted by basket cell axons while the axon hillocks and initial segments of granule cells were never contacted. These findings are consistent with previous immunocytochemical, and physiological data that indicate feedback inhibitory mechanisms in the dentate gyrus are mediated via mossy fibre collaterals which synapse with GABAergic basket cells. In addition, the electron microscopic data for basket cells are similar to those for aspinous stellate cells in the neocortex, another type of cortical, GABAergic local circuit neuron. Thus, the basket cells in the dentate gyrus may have a function similar to other inhibitory, cortical local circuit neurons.

The distribution of recurrent Purkinje collateral synapses in the mouse cerebellar cortex: an electron microscopic study.

AbstractMyelinated fibers found in the lower molecular layer of the mouse cerebellum form synaptic enlargements of a uniform morphological appearance. Similar synapses have been observed to come from myelinated fibers in the upper granularlower Purkinje cell layer. Both have been interpreted as Purkinje collateral boutons. The study of the distribution of Purkinje collateral synapses revealed that one third formed the supraganglionic plexus in the lower molecular layer and two thirds formed an infraganglionic plexus in the lower Purkinje upper granular zone. In the molecular layer, basket cell somata and dendrites received the gratest number of Purkinje collaterals, while Purkinje dendrites and branchlets received relatively few. In the infraganglionic plexus, the Purkinje collaterals terminated on “very low basket” cells and upon a dense dendritic “thicket” belived to be contributed largely by basket cells.