Clustering of L-type Ca2+ channels at the base of major dendrites in hippocampal pyramidal neurons.

Integration and processing of electrical signals in individual neurons depend critically on the spatial distribution of ion channels on the cell surface. In hippocampal pyramidal neurons, voltage-sensitive calcium channels have important roles in the control of Ca2(+)-dependent cellular processes such as action potential generation, neurotransmitter release, and epileptogenesis. Long-term potentiation of synaptic transmission in the hippocampal pyramidal cell, a form of neuronal plasticity that is thought to represent a cellular correlate of learning and memory, is dependent on Ca2+ entry mediated by synaptic activation of glutamate receptors that have a high affinity for NMDA (N-methyl(-D-aspartate) and are located in distal dendrites. Stimuli causing long-term potentiation at these distal synapses also cause a large local increase in cytosolic Ca2+ in the proximal regions of dendrites. This increase has been proposed to result from activation of voltage-gated Ca2+ channels. At least four types of voltage-gated Ca2+ channels, designated N, L. T and P, may be involved in these processes. Here we show that L-type Ca2+ channels, visualized using a monoclonal antibody, are located in the cell bodies and proximal dendrites of hippocampal pyramidal cells and are clustered in high density at the base of major dendrites. We suggest that these high densities of L-type Ca2+ channels may serve to mediate Ca2+ entry into the pyramidal cell body and proximal dendrites in response to summed excitatory inputs to the distal dendrites and to initiate intracellular regulatory events in the cell body in response to the same synaptic inputs that cause long-term potentiation at distal dendritic synapses.

Identification and localization of ryanodine binding proteins in the avian central nervous system.

Ryanodine binding proteins of the CNS have been identified using monoclonal antibodies against avian skeletal muscle ryanodine binding proteins. These proteins were localized to intracellular membranes of the dendrites, perikarya, and axons of cerebellar Purkinje neurons using laser confocal microscopy and immunoelectron microscopy. Ryanodine binding proteins were not found in dendritic spines. Immunoprecipitation and [3H]epiryanodine binding experiments revealed that the cerebellar ryanodine binding proteins have a native molecular weight of approximately 2000 kd and are composed of two high molecular weight (approximately 500 kd) polypeptide subunits. A comparable protein having a single high molecular weight polypeptide subunit was observed in the remainder of the brain. If the ryanodine binding proteins in muscle and nerve are similar in function, then the neuronal proteins may participate in the release of calcium from intracellular stores that are mechanistically and spatially distinct from those gated by inositol trisphosphate receptors.

Pathological proteins Tau 64 and 69 are specifically expressed in the somatodendritic domain of the degenerating cortical neurons during Alzheimer's disease. Demonstration with a panel of antibodies against Tau proteins.

Bundles of paired helical filaments (PHF) accumulate in the pyramidal neurons that degenerate during Alzheimer's disease. This neurofibrillary degeneration is highly correlated with clinical signs of dementia. During the degenerating process, Tau proteins, which are the major antigenic components of PHF, are abnormally phosphorylated and two pathological isoforms named Tau 64 and 69 are expressed. We have studied their immunoblot distribution in the cortical gray and white matter from different regions of normal and Alzheimer brains, to determine if the degenerating process preferentially affects the somatodendritic or the axonal domain. Two categories of antibodies were used. The first category consisted of anti-human native Tau, anti-Tau proteins from different vertebrates, anti-PHF, monoclonal antibody Alz-50 and an anti-C terminal repeated region of Tau. In control brains, these antibodies strongly detected normal Tau proteins in the gray matter while Tau immunodetection was weak in the white matter. In Alzheimer brain cortices, each antibody detected Tau 64 and 69 in gray matter extracts but not at all in white matter extracts. The second category of anti-Tau consisted of the anti-PHF saturated with normal brain protein extracts. This antiserum only probed the abnormally phosphorylated Tau proteins. It detected Tau 64 and 69 exclusively in the cortical gray matter of Alzheimer brains. Moreover, a 55-kDa Tau protein was also immunolabelled, which might be an intermediary form between normal Tau and Tau 64 and 69. Our results demonstrate that Tau proteins are normal and major components of the somatodendritic domain and that Tau pathology, reflected by the presence of Tau 64 and 69, affects preferentially this domain during Alzheimer's disease.

An 18-kd heparin-binding protein of developing brain that is distinct from fibroblast growth factors.

An 18-kd heparin-binding protein (p18) was isolated from perinatal rat brain. Although the protein closely resembles the fibroblast growth factors in its strong binding to heparin and in its apparent molecular mass, it has a distinct structure. This was concluded from the amino-terminal sequence analysis that identified a unique structure containing a cluster of lysine residues. Antipeptide antibodies were raised in rabbits according to the sequence analysis and affinity purified using a synthetic peptide. The antibodies were shown to bind specifically to p18, which was immunochemically distinct from the basic fibroblast growth factor. The antipeptide antibodies detected p18 in brain but not in liver, kidney, heart or skeletal muscle. The content of the protein was shown to undergo a remarkable developmental change corresponding to the time period of rapid sprouting of axons and dendrites in brain. The content of p18 was rapidly increased at the time of birth until the postnatal age of approximately 1 week, after which it was decreased to values less than 10% in young adults as compared to the content found in perinatal rats. p18 also enhanced neurite outgrowth in brain neurons in vitro. The protein was stained in neurons in cells dispersed from perinatal brain. The properties of p18 suggest that it has a role in the growth and maturation of brain.

Ultrastructure of primary afferent fibres and terminals expressing alpha-galactose extended oligosaccharides in the spinal cord and brainstem of the rat.

The ultrastructural characteristics of primary afferent fibres, which express alpha-galactose extended oligosaccharides recognized by LD2 and LA4 monoclonal antibodies, and the subcellular localization of these oligosaccharides were studied. LD2 and LA4 antibodies both label intensely the plasma membrane of primary afferent fibres, and with LD2 antibody all immunoreactive profiles also possessed strong intracellular staining. In contrast, intracellular staining with LA4 antibody was observed in only a subpopulation of stained profiles. LD2-immunoreactive fibres were detected in trigeminal and Lissauer tracts and in lamina I (LI) and lamina II (LII), and appeared as a mixture of unmyelinated and myelinated fibres. The highest density of LD2-immunoreactive synaptic boutons was found in lamina II outer (LIIo). Many of the terminals were simple dome-shaped terminals, making single asymmetric synapses over small and medium-sized dendritic shafts and dendritic spines. All LA4-immunoreactive fibres were unmyelinated. In addition, some small scalloped central-glomerular terminals contacting two or three dendrites were found. LA4-immunoreactive fibres were found more frequently than terminals and appeared most heavily immunostained in trigeminal and Lissauer tracts. In the neuropil of LI and LII, LA4 profiles were generally very weakly immunostained, although a small sample of immunostained synaptic boutons was detected. All LA4-immunoreactive terminals were found in lamina II inner (LIIi) and made simple asymmetric axodendritic synapses. In addition to axons and terminals, some dendrites exhibited LD2 immunoreactivity and this was most intense in the region of synaptic vesicles. In addition to neurons, some endothelial cells were immunostained with LD2 antibody and astrocytes were immunostained with LA4 antibody.

Ultrastructural localization of thyrotropin-releasing hormone immunoreactivity in the dorsal vagal complex in rat.

Thyrotropin-releasing hormone-like immunoreactivity (TRH-LI) was localized at the ultrastructural level in the dorsal vagal complex (DVC: dorsal motor nucleus of the vagus (DMV) and the nucleus of the solitary tract (NST] in rat. TRH-LI was concentrated in large granular vesicles in axons, presynaptic terminals, and non-synaptic axon varicosities. TRH-LI presynaptic terminals established both asymmetric and symmetric synaptic contacts with dendrites. These observations are consistent with recently described direct inhibitory and facilitatory effects of TRH on the electrical activity of neurons in the DVC.

Co-lamination of cholinergic amacrine cell and displaced ganglion cell dendrites in the chicken retina.

Displaced ganglion cells in the chicken retina were back-labelled with Fast blue injected into the nucleus of the basal optic root, then filled under visual control with Lucifer yellow to reveal the dendritic fields of the cells. In transverse sections, the dendrites of the displaced ganglion cells formed a narrow band in the outer part of the inner plexiform layer. Counter-staining for acetylcholinesterase (AChE) activity showed that the dendrites of the displaced ganglion cells and the type I cholinergic amacrine cells co-laminated in the outer part of the inner plexiform layer.

Alterations in Langerhans cells and Thy-1+ dendritic epidermal cells in murine epidermis during the evolution of ultraviolet radiation-induced skin cancers.

To understand the role of cutaneous immune cells in host resistance to the induction and growth of skin cancer, we investigated the number and morphology of murine dendritic epidermal cells (dEC) during the evolution of ultraviolet (UVA) UV-induced skin cancers. Female C3H/HeN mice were treated topically with 8-methoxypsoralen followed by ultraviolet A (UVA) radiation 3 times/week or irradiated with UVB radiation 3 times/week. In both psoralen plus UVA- and UVB-treated mice, ATPase+ and Ia+ Langerhans cells almost completely disappeared from the treated skin during the early latency period of tumor development (4 weeks) but reappeared in the epidermis late in the latency period (between 15 and 22 weeks). The ATPase+ cells that reappeared in the epidermis had a rounder, less dendritic morphology than normal Langerhans cells. Thy-1+ dEC were totally depleted from the epidermis in both treatment groups at the end of first week of treatment and were nearly absent from the skin during the entire latency period. After tumors appeared (29 weeks), Thy-1+ dEC were still absent or detected only in small numbers in skin surrounding the tumors. ATPase+ and Ia+ cells present in skin around the tumors constituted 60 to 80% of the number in nonirradiated skin. Mice that received UVA radiation alone developed no tumors. ATPase+ and Ia+ Langerhans cells and Thy-1+ dEC were detected in UVA-treated epidermis after 22 weeks and 43 weeks, although the numbers were lower than those in unirradiated mice. Most psoralen plus UVA-induced tumors (81%) were squamous cell carcinomas, whereas only 24% of UVB-induced tumors were of this histological type. Our results demonstrate that UV-induced skin cancers developed in the presence of ATPase+ and Ia+ cells in the epidermis and in the absence of Thy-1+ dEC.

Localization of GABA-like immunoreactivity in the ectostriatum of domestic chicks: GABA immunocytochemistry combined with Golgi impregnation.

The distribution of GABA-like immunoreactivity (GABA-LI) in the ectostriatal core (Ec) of domestic chicks (one to two days old) was investigated using (1) preembedding GABA immunocytochemistry and (2) Golgi impregnation and gold-toning combined with postembedding GABA immunocytochemistry. Two major classes of neurons which display GABA-LI were identified in chick Ec. Firstly, large GABA immunopositive cells which comprise at least two further subtypes: an ovoid or polygonal form of 14-18 microns diameter with no apparent polarity of dendrites and a smaller cell (10-14 microns) with ovoid or basket-shaped soma and often more polarized dendritic ramification. In both subtypes the dendritic surface is smooth or sparsely spiny. Secondly, a small GABA immunopositive cell which is characterized by a round cell body of 5-8 microns diameter and thin and sparsely ramifying dendrites of smooth surface or with irregular protrusions. Based upon comprehensive descriptions of ectostriatal cytoarchitectonics (Tömböl et al., 1988c), and synaptology (Watanabe et al., 1985), we argue that the GABA-immunopositive cell types of chick Ec are likely to represent inhibitory interneurons comparable with GABAergic inhibitory cell types described in mammalian visual cortex.

Gap junction protein in rat hippocampus: correlative light and electron microscope immunohistochemical localization.

Immunohistochemical techniques and an affinity-purified antibody directed against the 27-kD gap-junctional protein (GJP) from rat liver were used to determine the ultrastructural localization of GJP in the rat hippocampus. At the light microscope level, dense GJP immunoreactivity having a stringlike appearance was seen in a very small percentage of medium-sized neuronal somata located in the stratum pyramidale, and diffuse immunostaining was seen in many small cell bodies in the stratum pyramidale, stratum oriens, and the alveus. Abundant GJP-immunoreactive (GJP-IR) varicose fibers were observed in the strata pyramidale, radiatum, and oriens but were less concentrated in the alveus. Numerous punctate GJP-IR elements were observed in all hippocampal layers. Upon EM analysis, GJP-IR neuronal somata in the stratum pyramidale were found to be, without exception, nonpyramidal neurons as judged by such distinguishing features as their fusiform perikarya, indented nucleus, and well-developed rough endoplasmic reticulum (RER). Immunostaining within these cells was largely localized to the Golgi apparatus and associated vesicular components. Small, diffusely GJP-IR cells were identified ultrastructurally as protoplasmic and fibrous astrocytes. Immunostaining within these cells was localized to the Golgi apparatus, RER, and small, ribosomelike bodies 15-25 nm in diameter. Among neuronal processes GJP immunoreactivity was found within dendrites, axons, and axonal terminals. The latter structures contained numerous GJP-IR vesicles having an average diameter of about 40 nm. A frequent observation indicating some degree of specificity of the anti-GJP antibody employed here was immunostaining of typical gap junctions between dendrites and, more commonly, between processes of glial cells. Occasionally, however, GJP-IR dendrodendritic, axodendritic, and axoaxonic contacts were found that could be considered, at best, as being gap-junction-like (gj-L). In these cases, asymmetric immunostaining of adjacent plasma membranes forming gj-L structures was not uncommon. These results confirm the existence of gap junctions between dendrites in the rat hippocampus and demonstrate that GJP immunoreactivity on cytoplasmic membranes is restricted either to typical neuronal and glial gap junctions or to gj-L structures at circumscribed sites of contact between various types of neuronal elements where GJP may contribute to a novel mechanism of neural communication.

Morphologies of rabbit retinal ganglion cells with concentric receptive fields.

Rabbit retinal ganglion cells with concentric receptive fields were intracellularly recorded and stained in the isolated superfused eyecup preparation to relate specific physiological response properties to dendritic morphology. Concentric ganglion cells, as traditionally defined, were those that had On or Off centers with antagonistic surrounds but lacked complex response properties such as direction or orientation selectivity. Concentric cells were classified into different groups by extracellular recordings of their On- or Off-center response sign, excitatory receptive field center size, linearity of spatial summation, and brisk vs. sluggish and transient vs. sustained responses to step changes in light intensity. The cells were then impaled, confirmed in identity during intracellular recording, and iontophoretically injected with horseradish peroxidase for histological analysis. Twenty-three concentric ganglion cells were recovered and morphometrically analyzed. Their physiological response properties were found to be related to a number of underlying two- and three-dimensional attributes of the cell's dendritic branching patterns. The dendrites of all 20 brisk concentric cells and two of the three sluggish cells were found to ramify narrowly in either the proximal or distal half of the inner plexiform layer, corresponding to whether they are On center or Off center, respectively. One of the sluggish concentric cells was found to have a more complex, partially bistratified ramification. Physiologically identified brisk-sustained-linear, brisk-transient-nonlinear, brisk-transient-linear, and at least two classes of sluggish concentric ganglion cells were stained. Each of these physiological classes appears to exhibit a distinct and identifiable dendritic branching pattern.

Morphologies of rabbit retinal ganglion cells with complex receptive fields.

Ganglion cells that had complex receptive field properties, namely, On-Off and On direction-selective cells, orientation-selective cells, local edge detectors, and uniformity detectors (suppressed by contrast cells) were recorded in an isolated superfused rabbit eyecup preparation. Cells were first classified by their characteristic extracellular responses to manually controlled stimuli similar to those which have been used in previous in vivo studies. Ganglion cells were then impaled, confirmed in identity by intracellular recording, and iontophoretically injected with horseradish peroxidase for staining. Twenty-two ganglion cells, which included members of all the major classes mentioned above, were recovered from the visual streak or near periphery. All recovered cells were drawn in camera lucida from flat-mounted retinas and entered into a computer as two-dimensional stick figures; nearly all were three-dimensionally reconstructed to determine the level and manner of dendritic ramification in the inner plexiform layer (IPL). The location of ganglion cell dendrites in sublaminar regions of the IPL was found to be consistent with the hypothesis of a division of the IPL into excitatory On (proximal) and Off (distal) sublaminae, with some qualifications for particular classes. Each of the complex receptive field ganglion cell classes exhibited a distinctive three-dimensional dendritic arborization pattern uniquely associated with that physiological class.

In situ localization of myosin and actin in dendritic spines with the immunogold technique.

The in situ detection of macromolecules by means of immunoelectron microscopy provides information about their ultrastructural localization in cellular compartments. With this technique, we have demonstrated that the contractile proteins actin and myosin are both localized in dendritic spines at densities exceeding those of other neuronal compartments. Myosin was associated with actin filaments, with spine plasma membrane, and with membranes of the spine apparatus. Given the dynamic properties of actin and myosin, these data suggest that these proteins may be involved in the mechanism of synaptic plasticity in general and in morphometric change resulting from intense synaptic activation in particular.

Selective localization of messenger RNA for cytoskeletal protein MAP2 in dendrites.

For nerve cells to develop their highly polarized form, appropriate structural molecules must be targeted to either axons or dendrites. This could be achieved by the synthesis of structural proteins in the cell body and their sorting to either axons or dendrites by specific transport mechanisms. For dendrites, an alternative possibility is that proteins could be synthesized locally in the dendritic cytoplasm. This is an attractive idea because it would allow regulation of the production of structural molecules in response to local demand during dendritic development. The feasibility of dendritic protein synthesis is suggested both by the existence of dendritic polyribosomes and by the recent demonstration that newly synthesized RNA is transported into the dendrites of neurons differentiating in culture. However, to date there has been no demonstration of the selective synthesis of an identified dendrite-specific protein in the dendritic cytoplasm. Here, we use in situ hybridization with specific complementary DNA probes to show that messenger RNA for the dendrite-specific microtubule-associated protein MAP2 (refs 3-5) is present in dendrites in the developing brain. By contrast the mRNA for tubulin, a protein present in both axons and dendrites is located exclusively in neuronal cell bodies.

Postnatal development of cat hind limb motoneurons. III: Changes in size of motoneurons supplying the triceps surae muscle.

The postnatal changes of neuronal dimensions were studied in cat triceps surae motoneurons intracellularly labeled with horseradish peroxidase. Systematic correlations were observed in the analysis of single dendrites at each studied stage, from birth to 44-46 days post natum (d.p.n.) age, between size parameters intrinsic to the dendrites as the diameter of a 1st-order dendrite, the combined dendritic length, the dendritic membrane area, and the degree of branching. Some variability among samples was evident in each studied age group. The correlations were, however, sufficiently close to permit indirect estimations of both combined dendritic length and dendritic membrane area for larger samples of neurons from data on dendritic stem caliber. The total postnatal increase in dendritic membrane area was, on the average, 400%, i.e., from close to 100 X 10(3) microns2 to about 500 X 10(3) microns2. The corresponding increase in soma area amounted to 100%. Analysis revealed that there was a time lag between the increase in somatic and dendritic size. Thus, adult somatic dimensions were attained at age 44-46 d.p.n.; however, at this stage, the mean total dendritic membrane area was only about half of the adult value. The postnatal increase in size appeared to vary among neurons, yielding a wider neuronal size spectrum in the adult cat than that observed in kittens. The measured increase in size corresponded to a calculated average addition of dendritic membrane area of 3700 microns2/day from birth to 22-24 d.p.n. and from that stage to 44-46 d.p.n. of 2700 microns2 per day. Likewise, the increase in combined dendritic length could initially be as large as 1 mm/day down to 0.4 mm/day between 22-24 and 44-46 d.p.n., with a mean growth during the first 44-46 d.p.n. of 0.5 to 0.6 mm/day. The ratios of daughters to parent branch diameters (sigmadd1.5: dp1.5) and the dendritic trunk parameter (sigma d1.5) recorded along the proximodistal dendritic path distance revealed transient changes that might impact on the electrotonic properties of the dendrites during postnatal development. Computations from the measured changes in dendritic branch lengths and calibers indicated that if membrane and internal resistivity remain unaltered during postnatal development, the dendritic domain is electrotonically more compact in the newborn kitten than in the adult cat.(ABSTRACT TRUNCATED AT 400 WORDS)

Postnatal development of lamina III/IV nonpyramidal neurons in rabbit auditory cortex: quantitative and spatial analyses of Golgi-impregnated material.

We have studied the postnatal development of lamina III/IV spine-free nonpyramidal neurons in the auditory cortex of the New Zealand white rabbit. The morphology and dendritic branching pattern of single cells impregnated with a Golgi-Cox variant were analyzed with the aid of camera lucida drawings and three-dimensional reconstructions obtained with a computer microscope. Sample sizes of 20 neurons were obtained at birth (day 0), postnatal day (PD) 3, 6, 9, 12, 15, 21, and 30 days of age. Normative data were also available from PD-60 and young adult rabbits studied previously. At birth, lamina II-IV have not yet emerged from the cortical plate; immature nonpyramidal neurons at the bottom of the cortical plate (presumptive layer IV) are characterized by short, vertically oriented dendrites. Growth-cone-like structures are present along the shafts and at the tips of the dendrites. At birth, soma area and total dendritic length are, respectively, 34 and 10% of adult values. The cortical plate acquires a trilaminar appearance at PD-3. The six-layered cortex is present by PD-6. During the first postnatal week dendritic length increases fourfold and is accompanied by a significant increase in both terminal and preterminal dendritic growth cones. At the onset of hearing at PD-6, there is a significant proliferation of dendrites and branches to 144 and 200% of adult levels, respectively. These supernumerary dendrites are rapidly lost during the second postnatal week, at which time the somata and dendrites become covered with spines. The loss of higher-order dendrites occurs more gradually; the number of dendritic branches is still 116% of adult values at PD-30. Spine density peaks between days PD-12 and PD-15, and then gradually diminishes until the cells are sparsely spined or spine free by PD-30. Total dendritic length increases in a linear fashion up to PD-15, at which time it is 80% of adult values. An analysis of terminal and intermediate branches demonstrated that the increase in total dendritic length after PD-6 is due entirely to the growth of terminal dendrites. Total dendritic length attains adult levels by PD-30. Spatial analyses revealed that a vertical orientation of dendrites is present at birth. Associated with the onset of hearing at PD-6, there is an explosive elaboration of dendrites toward the pial surface.(ABSTRACT TRUNCATED AT 400 WORDS)

A Golgi analysis of the gustatory zone of the nucleus of the solitary tract in the adult hamster.

The somal shapes, dendritic features, and orientations of the neurons within the gustatory zone of the nucleus of the solitary tract were studied with the rapid Golgi method in the adult hamster. These Golgi studies complement previous quantitative morphometric analyses of the distributions of large and small neurons within the gustatory zone. Class 1 neurons are usually fusiform and possess long, relatively unbranched dendrites that often extend beyond the cytoarchitectonic boundaries of the gustatory zone. Class II neurons are multipolar and possess more dendrites that are significantly shorter than those of class I neurons. Both classes of neurons are spine poor. Computer-generated three-dimensional rotational analyses demonstrate that the dendritic arborizations of neurons of the gustatory zone are oriented preferentially in the horizontal plane. Dendrites extend in parallel or perpendicular to the solitary tract, the source of peripheral gustatory inputs, and appear to be positioned spatially to maximize synaptic interactions with these peripheral fibers. These Golgi studies also suggest that individual gustatory neurons may be influenced by incoming gustatory fibers that innervate separate populations of taste buds, a finding that is not predictable from the topographical organization of the gustatory zone.

Electrophysiological characteristics of morphologically identified reticular thalamic neurons from rat slices.

This study is aimed at the investigation of the morphological and electrophysiological characteristics of neurons from the nucleus reticularis thalami in rat thalamic slices incubated in vitro. Ten neurons were recorded in the ventrobasal complex, four of which were successfully injected following horseradish peroxidase injection. Two main types of reticular thalamic neurons were morphologically identified: (1) the small fusiform 'f' cells characterized by a very elongated perikaryon, dendritic arborization prevalent in the rostrocaudal and dorsoventral planes, and an axon without any collaterals branching within the nucleus reticularis thalami; and (2) the large fusiform 'F' neurons with dendrites arborizing mainly in the horizontal plane and with axonal branches within the nucleus reticularis thalami. The electrophysiological properties of the neurons were similar in F and f cells. The reticular neurons showed, in resting conditions, a single spike response followed by a postexcitatory hyperpolarizing potential. The hyperpolarization of these neurons transformed the single spike response into a burst discharge similar to that observed in thalamic relay neurons at resting membrane potential. The same phenomenon was observed when bicuculline was administered by perfusion to the slices and, in this case, a recovery to a single spike response was obtained by a depolarizing d.c. current injection. By contrast, the local administration of GABA induced a depolarization with a pronounced decrease in input resistance. The present data demonstrate the presence of at least two neuronal subtypes within the nucleus reticularis thalami, suggesting that only one is responsible for the phenomenon of auto-inhibition by means of intrinsic axon collaterals. Moreover, it is hypothesized that intranuclear GABAergic collaterals could control neuronal excitability of reticular thalamic cells by both shunting the membrane and shifting the burst firing to a single spike firing mode.