Isolectin B4 (IB4)-conjugated streptavidin for the selective knockdown of proteins in IB4-positive (+) nociceptors.

In vivo analysis of protein function in nociceptor subpopulations using antisense oligonucleotides and short interfering RNAs is limited by their non-selective cellular uptake. To address the need for selective transfection methods, we covalently linked isolectin B4 (IB4) to streptavidin and analyzed whether it could be used to study protein function in IB4(+)-nociceptors. Rats treated intrathecally with IB4-conjugated streptavidin complexed with biotinylated antisense oligonucleotides for protein kinase C epsilon (PKCε) mRNA were found to have: a) less PKCε in dorsal root ganglia (DRG), b) reduced PKCε expression in IB4(+) but not IB4(-) DRG neurons, and c) fewer transcripts of the PKCε gene in the DRG. This knockdown in PKCε expression in IB4(+) DRG neurons is sufficient to reverse hyperalgesic priming, a rodent model of chronic pain that is dependent on PKCε in IB4(+)-nociceptors. These results establish that IB4-streptavidin can be used to study protein function in a defined subpopulation of nociceptive C-fiber afferents.

Schwann cells are removed from the spinal cord after effecting recovery from paraplegia.

Remyelination of the CNS is necessary to restore neural function in a number of demyelinating conditions. Schwann cells, the myelinating cells of the periphery, are candidates for this purpose because they have more robust regenerative properties than their central homologs, the oligodendrocytes. Although the ability of Schwann cells to remyelinate the CNS has been demonstrated, their capacity to enter the adult spinal cord in large numbers and effect functional recovery remains uncertain. We used cholera toxin B-subunit conjugated to saporin to demyelinate the rat lumbar spinal cord, remove macroglia, and produce paraplegia. After the removal of oligodendrocyte and astrocyte debris by invading macrophages, there was a spontaneous entry of Schwann cells into the spinal cord, along with axonal remyelination and concomitant functional recovery from paraplegia occurring within 75 d. The Schwann cells appeared to enter the dorsal funiculi via the dorsal root entry zone and the lateral funiculi via rootlets that had become adherent to the lateral spinal cord after the inflammation. In the following weeks, Schwann cell myelin surrounding central axons was progressively replaced by oligodendrocyte myelin without lapse in motor function. Our results show that endogenous Schwann cells can reverse a severe neurological deficit caused by CNS demyelination and enable later oligodendrocyte remyelination.

Quantitative analyses of intracellularly characterized and labeled thalamocortical projection neurons in the ventrobasal complex of primates.

This study describes the architecture of neurons and individual dendritic arbors of thirteen intracellularly labeled thalamocortical projection neurons that respond to non-noxious stimuli from the primate (Macaca fascicularis or Macaca mulatta) ventrobasal complex (VB). The neurons compose a homogeneous morphological class with total dendritic lengths from 10,169 microns to 21,711 microns (mean 17,615 microns +/- 3,705). The labeled neurons were remarkably similar in most measured parameters including the number of dendrites (7.5 +/- 1.2), percentage of dichotomous branching (89.8% +/- 3.4), and contribution of terminal branches to total dendritic length (88.4% +/- 2.0). The individual dendrites ranged in total length from 443 microns to 7,657 microns with a mean of 2,346 microns (+/- 137, n = 98). There was a positive correlation between stem dendrite diameter and total dendrite length, making it possible to estimate the total size of an individual dendrite by measuring the stem dendrite diameter. There was only a small increase in mean path distance with increasing dendritic size at the whole neuron and individual dendritic levels, so that for individual dendrites the mean path distance of a dendrite consisting of only two segments was 199 microns, while the mean path distance for a dendrite with eight segments was only 45 microns longer. Analysis of dendrite diameter, segment order, and path distance shows that dendritic diameter is not reliable for determining the location of synaptic contacts viewed by electron microscopy onto dendritic trees. The small variation of measured parameters between these neurons presents a powerful tool for future developmental, plasticity and comparative studies of VB neurons.

Quantitative analysis of the dendrites of sacral preganglionic neurons in the cat.

Quantitative analyses were performed on the dendrites and somata of 25 electrophysiologically identified preganglionic neurons (PGN) obtained from the sacral spinal cord of the cat by intracellular injection of Neurobiotin or horseradish peroxidase. Total dendritic length and surface area were measured for each dendrite. The sizes of the stem dendrites measured at their base were positively correlated with the sizes of the entire tree and numbers of end branches. Total surface area of somata and dendrites averaged 39,138 microm(2); 90.7% of that was from the dendrites. To obtain measurements of the relative contributions of PGN dendrites to specific regions of the spinal cord, the percentage of each dendrite occupying eight spinal cord regions was recorded. Sixty-three percent of the dendrites projected dorsal to their somata, whereas an average of 33.3% of dendrites were located in the white matter, most of them in the lateral and dorsolateral funiculi. The neurons within this sample formed a continuum with some neurons having a large percentage of dendrites in lamina I but little in the white matter, whereas at the other end of the continuum were cells with the reverse configuration. The intermediate neurons had dendrites in both locations. Taken together, these data indicate a heterogeneous population of PGN in the lateral band of the sacral parasympathetic nucleus.

Dendritic architecture of rat somatosensory thalamocortical projection neurons.

This study examines dendrites from physiologically characterized and intracellularly labelled thalamocortical projection (TCP) neurons from the rat ventrobasal complex (VB) and posterior nucleus (POm). The goals were to provide quantitative descriptions of TCP neuron dendrites, examine underlying design principles of dendritic morphology, and determine correlations between dendritic size parameters. Forty-four dendrites from seven VB neurons and 21 dendrites from three POm TCP neurons that responded to low-threshold mechanical stimuli were reconstructed and quantitatively analyzed at the light microscopic level. The dendritic architecture of the neurons was remarkably similar in most parameters studied, including the percentage of dichotomous branching, contribution of terminal branches to total dendritic length, and branching symmetry. There was a positive correlation between stem dendrite diameter and the length of the entire dendrite arbor, making it possible to estimate the total length of a dendritic arbor by measuring the stem dendrite diameter. The correlations of the VB and POm dendrites had different slopes. The path distance (the distance from the soma to a dendritic end point) of individual dendrites showed only a small variation with large differences in the total dendritic length of an arbor. The constant diameter of distal dendrites shows that dendrite diameter is a poor predictor of synaptic location on the dendritic tree. Although the morphology of neurons and their individual dendrites varied considerably in overall size and qualitative appearance, when examined qualitatively, many aspects of dendritic structure were similar within and between groups. We suggest that the rat somatosensory TCP neurons have a stereotyped dendritic architecture and present data which provide a base for future comparative, developmental, and plasticity studies.

Ultrastructural analysis of GABA-immunoreactive elements in the monkey thalamic ventrobasal complex.

This study describes the ventrobasal complex of the primate by using GABA immunocytochemistry at the electron microscopic level. The primate ventrobasal complex has a similar synaptic organization to sensory thalamic nuclei in other species. Two synaptic profiles within the ventrobasal complex contain flattened or pleomorphic synaptic vesicles and are GABA-immunoreactive. F-boutons (= F1 type, Guillery's classification; Guillery: Z. Zellforsch. 96:1-38, '69) are located principally in the extraglomerular neuropil and contain densely packed flattened synaptic vesicles and several elongate mitochondria and establish symmetric (Gray's type II) synaptic contacts. These boutons are not found postsynaptic to any other element and are presynaptic principally to nonimmunoreactive elements that are thought to be thalamocortical relay cell dendrites. PSD-boutons (= F2 type, Guillery's classification) contain a moderate number of flattened or pleomorphic synaptic vesicles and fewer mitochondria than F-boutons. PSD-boutons are found in glomerular and extraglomerular areas of neuropil and establish symmetric synaptic contacts. These boutons are considered to be appendages of interneuron dendrites and are postsynaptic to RL-, RS (Guillery's classification)-, F-, and other PSD-boutons. PSD-boutons are presynaptic to thalamocortical relay neurons and interneuron dendrites including PSD-boutons. Problems in distinguishing F- from PSD-boutons are addressed by comparing immunostained and nonimmunostained material and by the use of serial sections. The majority of synaptic contacts between pleomorphic vesicle-containing profiles appear to be between PSD-boutons and other components of interneurons. Few contacts between F-boutons and local circuit neurons are seen. These data suggest the principal GABAergic input to interneurons in the primate ventrobasal complex is derived from other interneurons.

Are there connections between the thalamic reticular nucleus and the brainstem reticular formation?

Increasing awareness that the thalamic reticular nucleus (TRN) plays an important role in controlling the output of cortically projecting cells in nuclei of the dorsal thalamus has focused attention on the question of whether there exist ascending projections to the TRN from the mesencephalic or other parts of the brainstem reticular formation (BRF). We have examined this and the related question of whether the neurons of TRN project to the BRF, by anterograde and retrograde tracing experiments with horseradish peroxidase (HRP) and HRP conjugated to wheat germ agglutinin. Injections of tracer were placed stereotaxically in the BRF at various depths and rostrocaudal and mediolateral coordinates, and the TRN and adjacent nuclei were examined in serial coronal sections, using tetramethylbenzidine as the principal chromogen. Retrogradely labelled cell bodies were consistently seen in hypothalamus and zona incerta but never in TRN, suggesting that, in the rat, TRN neurons do not project caudal to the thalamus. After 54 out of 60 injections, no terminal label was detected in any part of the TRN although such label was present in other parts of the thalamus, including the intralaminar nuclei, in the same sections. We therefore conclude that direct projections from the BRF to the TRN must be extremely sparse, and that those effects of BRF stimulation upon thalamocortical transmission that are mediated by the TRN (rather than by direct projections to dorsal thalamic nuclei) probably depend chiefly on indirect polysynaptic pathways.

Architecture of individual dendrites from intracellularly labeled thalamocortical projection neurons in the ventral posterolateral and ventral posteromedial nuclei of cat.

This study provides quantitative descriptions of individual dendrites from electrophysiologically characterized and intracellularly labeled thalamocortical projection (TCP) neurons of the cat ventrobasal complex. One hundred nine dendrites from six ventral posterolateral (VPL) neurons and six ventral posteromedial (VPM) neurons were examined. Measurement of several parameters showed that the individual dendrites were very similar to each other in overall architecture even though they varied greatly in total length and number of dendritic branches. The mean path distance (length from soma to a dendritic tip) was very similar for all dendrites in each group (VPL or VPM) regardless of the number of branches found along the path distance. However, VPL dendrites had a longer mean path distance (VPL = 206 +/- 36 microns; n = 51) than VPM dendrites (VPM = 182 +/- 29 microns; n = 58; P < 0.001). For all dendrites there was a strong correlation between the stem dendrite diameter and the dendritic length, which allows the estimation of dendritic length from dendrite diameter. Analysis of dendritic scaling shows that branches higher than first order do not follow Rall's 3/2 power rule, so these neurons cannot be modeled using the equivalent cylinder approximation. The data add to the qualitative descriptions of cat ventrobasal (VB) TCP dendrites currently available and provide a basis for future comparative, developmental, and plasticity studies. Analysis shows that many parameters of cat VB TCP dendrites fall within a narrow range, suggesting that, regardless of differences in length or superficial appearance, these dendrites share a stable underlying architecture.

Vasoactive intestinal polypeptide in sacral primary sensory pathways in the cat.

Unmyelinated sensory axons in the sacral spinal cord may play a role in bladder reflexes under certain pathological conditions. Previous data suggested vasoactive intestinal polypeptide (VIP) might be contained exclusively in sensory C-fibers, some of which innervate the bladder. This study was undertaken to describe the morphology of these VIP fibers in the sacral cord of the cat. VIP immunoreactivity was confined to unmyelinated axons observed at several levels of the sensory pathway including the dorsal root ganglia, dorsal roots, Lissauer's tract, and the lateral collateral pathway. A combination of light and electron microscopic observations showed VIP-immunoreactive fibers with labeled varicosities and synaptic terminals in laminae I, IIo, V, VII, and X. VIP-immunolabeled varicosities had a mean diameter of 1.6 microm (range = 0.11-7.4 microm, S.D. = 1.01, n = 311) with a small percentage (8%) being relatively large (3-7.4 microm). VIP varicosities contained a mixture of small clear vesicles (CLV) and large dense core vesicles (LDV). Although most varicosities contained a moderate number of LDVs (14.86 LDVs/microm2), some varicosities contained a large number of LDVs, whereas others contained very few. Varicosities that possessed synaptic specializations were classed as terminals and were divided into three morphological classes. Two of these resembled Gray's Type I terminal, whereas a third was similar to the Gray's Type II terminal. There was no consistent relationship between vesicle content of the terminal and the type of synaptic contact it possessed. This study shows that in the sacral spinal cord of the cat, VIP terminals originate only from C-fibers, terminate primarily in laminae I and V, and exhibit a variety of morphologies consistent with heterogeneous origins and functions of the lower urinary tract.

Analysis of gamma-aminobutyric acidergic synaptic contacts in the thalamic reticular nucleus of the monkey.

Gamma-aminobutyric acidergic (GABAergic) neurons in the thalamic reticular nucleus (TRN) spontaneously generate a synchronous bursting rhythm during slow-wave sleep in most mammals. A previous study at the electron microscopic level in cat anterior TRN has suggested that synchronous bursting activity could result from the large number of presumably GABAergic dendrodendritic synaptic contacts. However, little is known about the synaptology of the monkey thalamic reticular nucleus and whether it contains dendrodendritic contacts. To address this issue, we examined tissue obtained from Macaca fascicularis that was prepared for electron microscopy using postembedding techniques to demonstrate GABA immunoreactivity. Examination of the anterior (motor) and posterior (somatosensory) portions of the TRN disclosed the following: The majority of synaptic contacts (87.5% of 958) were formed by axon terminals showing no GABA immunoreactivity and making asymmetric synaptic contacts on dendrites or cell bodies. A further 6.4% of synaptic contacts was composed of GABA-immunoreactive presynaptic terminals making symmetric contacts with the dendrites of TRN neurons. The majority resembled the pleomorphic vesicle containing F-terminals seen in the dorsal thalamus and known to originate from axons of TRN. A subset or possible second class did not resemble any previously described class of GABA-immunoreactive terminals in the TRN. Both classes of these terminals making symmetric contacts may originate wholly or partially within the nucleus. There was one dendrodendritic synaptic contact and only a small number (3.2%) of axodendritic contacts with synaptic vesicles visible both pre- and postsynaptically. We conclude that dendrodendritic contacts are probably not responsible for the synchronized bursting neuronal activity seen in the slow-wave sleep of monkeys, and that, if TRN neurons are coupled synaptically, the most likely mechanism is through the synapses formed by recurrent axon collaterals of TRN neurons onto TRN dendrites.

Transneuronal changes of the inhibitory circuitry in the macaque somatosensory thalamus following lesions of the dorsal column nuclei.

The inhibitory circuitry of the ventroposterolateral nucleus (VPL) of the macaque somatosensory thalamus was analyzed in normal animals and in those surviving for a few days or several weeks following a unilateral lesion of the cuneate nucleus, the source of medial lemniscal (ML) axons carrying information from the contralateral upper extremity. Inhibitory synaptic terminals in the VPL were defined as those that contain flattened or pleomorphic synaptic vesicles and that can be shown to be immunoreactive for gamma-aminobutyric acid (GABA). There are two types of these profiles: F axon terminals that arise from neurons of the thalamic reticular nucleus, and perhaps from VPL local circuit neurons (LCNs); and the dendritic appendages of LCNs that form presynaptic dendrites (PSDs). ML terminals normally have extensive synaptic interactions with PSDs but not with F axon terminals. Electron microscopic analyses revealed that cuneatus lesions resulted in a rapid loss of ML terminals and a statistically significant reduction in both F and PSD synaptic profiles. Confocal scanning microscopy also demonstrated a profound loss of GABA immunoreactivity in the deafferented VPL. These changes persisted for more than 20 weeks, without any evidence of reactive synaptogenesis of surviving sensory afferents or of inhibitory synapses. The changes in GABA circuitry are transneuronal, and the possible mechanisms that may underlie them are discussed. It is suggested that the altered GABAergic circuitry of the VPL in the monkey may serve as a model for understanding changes in somatic sensation in the human following peripheral or central deafferentation.

Noradrenergic input to nociceptive modulatory neurons in the rat rostral ventromedial medulla.

Within the rostral ventromedial medulla (RVM), there are two classes of putative pain modulation neurons: ON cells and OFF cells, which respectively burst or pause prior to withdrawal reflexes elicited by noxious stimulation. Alpha-adrenergic agonists injected into the RVM produce changes in the latency of spinal nocifensive reflexes and, when iontophoretically applied, alter the firing of RVM ON but not OFF cells. To provide further information about the contribution of norepinephrine to RVM neuron function, we analyzed the distribution of tyrosine hydroxylase immunoreactive (TH-ir) appositions upon RVM ON and OFF cells. In the lightly anesthetized rat, seven ON and five OFF cells were identified by changes in their discharge rate in relation to nociceptive withdrawal reflexes and were labeled by intracellular injection of neurobiotin. Sections containing labeled cells were visualized by using avidin conjugated to a Texas Red fluorophore. Tissue with labeled cells was subsequently processed for TH-ir by using a Bodipy fluorophore conjugated secondary antibody. The distribution of the Bodipy-labeled fibers and terminals upon the Texas Red-labeled neurons was mapped using a confocal laser-scanning microscope. All the labeled neurons exhibited close TH-ir appositions. Appositions were of two types: swellings and fibers. Although the numbers and density of appositions varied among the cells, there were no consistent differences that correlated with physiological properties. Thus the overall density of appositions for ON cells (29.0 +/- 22.2 x 10(4) microns2) did not differ significantly from that for OFF cells (25.4 +/- 22.2 x 10(4) microns2). Tyrosine hydroxylase immunoreactive (TH-ir) appositions upon ON and OFF cells varied with their location along the dorso-ventral axis with more ventral neurons having a greater density of TH-ir swelling-type appositions. In a separate study, TH-ir and dopamine-beta-hydroxylase-like immunoreactivity (DBH-ir) were mapped in the same sections by using confocal microscopy. Nearly 97% of the TH-ir profiles co-localized with DBH-ir. These observations provide evidence that both ON and OFF cells in the RVM are targeted by noradrenergic inputs.

GABA puts a stop to pain.

A lack of inhibition, particularly that mediated by gamma-amino butyric acid (GABA), the main inhibitory transmitter of the central nervous system (CNS), is responsible for many pain states. Until recently, few GABA acting drugs were available and were prescribed mostly for relieving muscle spasms, anxiety and epilepsy, but rarely for pain. The basic metabolic pathway of GABA is well known and we are now beginning to understand the function of this neurotransmitter in the complex circuitry underlying pain, especially in the context of nerve injury. Analgesic compounds are now being developed targeting GABA transporters as well as GABA associated enzymes and receptors. Some GABA analogs act by inhibiting ion channels, a property that contributes to their analgesic effects. However, despite considerable progress in developing new compounds, the use of systemically acting GABAergic drugs is limited by unwanted side-effects on systems other than those involved in pain, and by the fact that in certain areas of the brain, GABA can enhance rather than reduce pain. The advent of new drugs targeting subtypes of GABA receptors and transporters and the possibility of using newly developed delivery systems, such as intrathecal pumps and viral vectors, to target specific areas of the nervous system will likely help circumvent these problems.

Fine structure of the dorsal part of the nucleus submedius of the rat thalamus: an anatomical study with reference to possible pain pathways.

The dorsal portion of the nucleus submedius of the rat thalamus receives spinal and trigeminal projections which may convey noxious inputs. The present study was undertaken to analyse the fine structure of the nucleus with particular reference to a possible trigemino-thalamo-prefrontal cortical pathway relaying in nucleus submedius. Presynaptic terminals in the dorsal portion of the nucleus submedius were classified into three broad categories usually observed in thalamic nuclei: "small round", "flat" and "large round" types. Axonal tracing using either anterograde transport of horseradish peroxidase or degeneration methods indicated that some "small round" terminals originate from the pre-frontal cortex. Some "large round" terminals were labelled from the trigeminal subnucleus caudalis. These "large round" terminals exhibited distinct morphological features when compared with trigeminal terminals in other thalamic nuclei. In particular they made synaptic contacts predominantly with dendritic protrusions and were surrounded by multilamellate astrocytic processes. Double-labelling experiments were performed by means of the combined retrograde transport of horseradish peroxidase and Wallerian degradation techniques. Terminals degrading after lesion of the trigeminal subnucleus caudalis contacted submedius neurons labelled retrogradely from the prefrontal cortex. These observations demonstrate the existence of a direct monosynaptically relayed pathway from subnucleus caudalis to prefrontal cortex which relays in the dorsal part of nucleus submedius.

Neural elements containing glutamic acid decarboxylase (GAD) in the dorsal lateral geniculate nucleus of the rat; immunohistochemical studies by light and electron microscopy.

Immunoreactive constituents of the dorsal lateral geniculate nucleus of adult albino rats were examined by light- and electron-microscopy, using the unlabelled antibody enzyme method, following treatment of brain slices with a purified antibody to glutamic acid decarboxylase. The neuropil of the dorsal lateral geniculate nucleus displayed a conspicuous granular immunoreactivity. In addition, the antibody was bound to a class of small neurons of characteristic morphology. These cells possessed few (commonly 2-4) sparsely branched, long dendrites from some of which immunoreactive appendages were traced. Many cells were bipolar in form, and the dendrites of some appeared to be preferentially orientated. The immunoreactive cells closely resembled intrinsic interneurons characterized in previous Golgi studies of this nucleus. By electron-microscopy, immunoreactive presynaptic elements were present both in the extraglomerular neuropil and in the synaptic glomeruli. The former were axon terminals containing flattened synaptic vesicles and making Gray type II axo-dendritic synaptic contact; they appeared to correspond to axon terminals whose origin in the thalamic reticular nucleus has been established in previous studies, but it is possible that some were axon terminals of intrinsic interneurons. The immunoreactive glomerular components also contained flattened vesicles, were presynaptic to presumptive projection cell dendrites, postsynaptic to retinal axon terminals, and participated in triplet (triadic) and other complex synaptic arrangements. They corresponded in all respects to the synaptic portions of the complex dendritic appendages of intrinsic interneurons, identified and characterized in previous studies. The finding that there are high levels of glutamic acid decarboxylase in the cell bodies, dendritic shafts and dendritic appendages of intrinsic interneurons in the dorsal lateral geniculate nucleus of the rat, and in the axon terminals of fibres projecting to this site from the thalamic reticular nucleus, allows us to conclude that the inhibitory inputs to the geniculo-cortical projection cells from both of these sources are probably mediated by gamma-aminobutyric acid.

Long-term intrathecal catheterization in the rat.

We report an intrathecal (i.t.) catheter system that permits repeated administration of volumes of 10 microl or more in the awake rat over many months. A small skin incision is made and a 32 ga polyurethane catheter is inserted in the sacral subarachnoid space using a modified 22 ga needle. The other end of the catheter is tunneled subcutaneously to the flank and exteriorized through a titanium port. The device is well tolerated, does not cause sensory or motor deficits, and does not interfere with behavioral testing. Rats equipped with this device can be housed with other rats. Over the 9 month observation period the function of the catheter was verified by repeated injection of 15 microl of 2% lidocaine that caused temporary paraplegia. Out of 12 implanted rats, the number of fully functional catheters was 10 at 3 months, seven at 6 months, and six at 9 months. At 3 months, i.t. injection of anti-dopamine-beta-hydroxylase antibodies conjugated to saporin (DBH-SAP, 5 microg/10 microl) resulted in noradrenergic denervation of the spinal cord in all rats (n=10). We propose that intrathecal catheterization is well suited for long term behavioral and pharmacological studies.

Convergent projections of trigeminal afferents from the principal nucleus and subnucleus interpolaris upon rat ventral posteromedial thalamic neurons.

Synaptic boutons originating from the trigeminal principal sensory nucleus (PrV) and the subnucleus interpolaris (SpI) which contact rat ventral posteromedial (VPM) neurons are similar in appearance. They are large boutons and contain a moderate packing density of round synaptic vesicles and established asymmetric (Gray type I) synaptic contacts principally on dendrites occasionally on somata. These boutons are similar to RL boutons of dorsal column nuclei and spinothalamic tract origin found in the ventral posterolateral thalamic nucleus. The boutons of PrV and SpI origin have an overlapping distribution on proximal dendrites of VPM neurons. Double labeling using degeneration and WGA-HRP shows that boutons from PrV and SpI contact the same VPM neurons confirming there is convergence of trigeminal afferents in VPM.

Thalamic reticular nucleus: anatomical evidence that cortico-reticular axons establish monosynaptic contact with reticulo-geniculate projection cells.

Cell bodies and dendrites of neurones in the thalamic reticular nucleus projecting to the dorsal lateral geniculate nucleus in adult albino rats, were labelled by retrograde transport of horseradish peroxidase (HRP). In the same animals the terminals of cortico-reticular axons were made identifiable by ipsilateral visual cortex ablation. Degenerating axon terminals established Gray type I synaptic contact with HRP-filled dendrites, indicating a monosynaptic connection between cortico-reticular and reticulo-geniculate projection cells.

Preserved features of thalamocortical projection neuron dendritic architecture in the somatosensory thalamus of the rat, cat and macaque.

A number of studies have shown that the organization of the mammalian somatosensory thalamus varies between species. As differences in cellular and synaptic thalamic organization would be expected to influence neuronal dendritic architecture, we compared somatosensory thalamocortical projection (TCP) neurons from the rat, cat and macaque. The results show that key features of the dendritic branching pattern remain unchanged despite large differences in the size of TCP neurons between the species. The features examined were: (i) ratio of the length of terminal branches to the length of the entire dendritic tree; (ii) the percentage of branch points that gave rise to two daughter branches as opposed to those that gave rise to three or more daughter branches; (iii) the proportional sum of absolute deviations (a measure of branching symmetry), and (iv) the mean branch order of the terminal segments. The present study provides evidence that somatosensory TCP neurons in these species comprise a homogeneous class and share a common dendritic architecture that is conserved across species despite changes in other aspects of thalamic circuitry. This suggests that TCP neuronal form is based on relatively stable genetic blueprint and that epigenetic factors (e.g. synaptic input) resulting from evolutionary changes in thalamic organization have had less influence on dendritic architecture.

Cell body and dendritic tree size of intracellularly labeled thalamocortical projection neurons in the ventrobasal complex of cat.

Fifteen thalamocortical projection (TCP) neurons from the adult cat ventrobasal complex (VB) were intracellularly labeled with horseradish peroxidase or neurobiotin and examined quantitatively. We find that cat TCP neurons share key morphological features and form one neuronal type. Previously reported variations in dendritic appearance cannot be supported by the present quantitative data. The number of dendrites varied between 4 and 13 (mean 9.1; +/- 4.0) and the total dendritic length of adult cat VB neurons varied between 9,421 and 19,646 microns (mean 13,120 microns; +/- 2,605). Linear regression analyses showed that soma diameter or cross-sectional area measurements provide a poor estimate for total dendritic length in TCP neurons. In contrast, the number of first order dendrites or the sum of first order dendrite diameters do provide a good estimate of overall TCP neuron size. This relationship is useful in predicting total dendritic length when it is not possible to reconstruct the entire dendritic tree. The mean dendritic path distance (distance from soma to the dendritic tip measured along the dendrite) was relatively constant for all neurons regardless of differences in total dendritic length or the number of branches that form the path distance.