Characteristics and surgical outcomes for medial temporal post-traumatic epilepsy.

A common post-traumatic location of epileptogenesis is the medial temporal lobe despite evidence of associated diffuse or remote cerebral injury. We undertook a review of post-traumatic medial temporal lobe epilepsy (MTLE) patients as part of an overall post-traumatic epilepsy population to assess the extent of cerebral injury sustained by this subpopulation and to establish whether surgical outcome differed from that of a non-traumatically-induced epilepsy population. A retrospective review of 57 patients operated for post-traumatic epilepsy (PTE) over a 10-year period (1993-2003) was undertaken with particular attention to those undergoing medial temporal resection. Preoperative magnetic resonance imaging (MRI) was assessed for the type and location of abnormalities. Postoperative outcomes were compared with those of patients with MTLE of non-traumatic origin operated by the same surgeon. Of the 57 patients operated, 30 cases underwent medial temporal lobe resection. The most common mechanism of injury was blunt trauma attributable to motor vehicle accidents with imaging abnormalities characterized by medial temporal sclerosis (MTS; 16 cases), T2/FLAIR hyperintensities (nine cases), periventricular gliosis (seven cases), diffuse cerebral atrophy (five cases) and focal encephalomalacia (three cases). Six patients had normal MRI studies. No significant differences in postoperative outcomes were found between post- and non-traumatic MTLE epilepsy groups. The presence of histopathological change in the medial temporal lobe varied greatly and provided no indication of a favourable postoperative outcome. Patients with post-traumatic medial temporal lobe epilepsy respond favourably to surgical treatment. In the case of medial temporal sclerosis, there is substantial variation of histopathological findings which correlate poorly with current imaging applications. The favourable outcomes obtained following surgery in this group attest to a commonality with other risk factors in the genesis of epilepsy in this location.

Auditory cortex stimulation for tinnitus.

Functional imaging techniques have demonstrated a relationship between the intensity of tinnitus and the degree of reorganization of the primary auditory cortex. Studies in experimental animals and humans have revealed that tinnitus is associated with a synchronized hyperactivity in the auditory cortex and proposed that the underlying pathophysiological mechanism is thalamocortical dysrhythmia; hence, decreased auditory stimulation results in decreased firing rate, and decreased lateral inhibition. Consequently, the surrounding brain area becomes hyperactive, firing at gamma band rates; this is considered a necessary precondition of auditory consciousness, and also tinnitus. Synchronization of the gamma band activity could possibly induce a topographical reorganization based on Hebbian mechanisms. Therefore, it seems logical to try to suppress tinnitus by modifying the tinnitus-related auditory cortex reorganization and hyperactivity. This can be achieved using neuronavigation-guided transcranial magnetic stimulation (TMS), which is capable of modulating cortical activity. If TMS is capable of suppressing tinnitus, the effect should be maintained by implanting electrodes over the area of electrophysiological signal abnormality on the auditory cortex. The results in the first patients treated by auditory cortex stimulation demonstrate a statistically significant tinnitus suppression in cases of unilateral pure tone tinnitus without suppression of white or narrow band noise. Hence, auditory cortex stimulation could become a physiologically guided treatment for a selected category of patients with severe tinnitus.

In vivo growth of C6 glioma cells transfected with connexin43 cDNA.

In order to examine the possible role of intercellular communication via gap junctions in the control of tumor growth, we have transfected C6 glioma cells with connexin43 cDNA. We obtained several clones with variable expression of connexin43. The growth rate of these clones in culture was inversely related to the degree of expression of the transfected cDNA. To examine the growth of these transfected cells in vivo, cells were grown in spinner culture flasks to form spheroids 250-300 microns in diameter. Spheroids of nontransfected C6 cells produced large gliomas. Immunohistochemical and in situ hybridization analyses revealed relatively high levels of connexin43 protein and mRNA in the host tissue, while little of this protein was detected in the glioma. In contrast, spheroids of connexin43-transfected cells grew more slowly and exhibited elevated levels of connexin43 protein and mRNA. These findings suggest that the expression of connexin43 may be associated with the control of brain tumor growth in vivo.

SPARC: a signal of astrocytic neoplastic transformation and reactive response in human primary and xenograft gliomas.

In an attempt to identify genetic alterations occurring early in astrocytoma progression, we performed subtractive hybridization between astrocytoma and glioblastoma cDNA libraries. We identified secreted protein acidic and rich in cysteine (SPARC), a protein implicated in cell-matrix interactions, as a gene overexpressed early in progression. Northern blot and immunohistochemical analyses indicated that transcript and protein were both elevated in all tumor specimens (grades II-IV) examined when compared with levels in normal brain. The level of SPARC expression was found to be tumor-dependent rather than grade-related. Immunohistochemically, SPARC protein was found to be overexpressed in 1) cells in the less cellularly dense regions within the tumor mass, 2) histomorphologically neoplastic-looking cells in adjacent normal brain at the tumor/brain interface, 3) neovessel endothelial cells in both the tumor and adjacent normal brain, and 4) reactive astrocytes in normal brain adjacent to tumor. Using a combination of DNA in situ hybridization and protein immunohistochemical analyses of the human/rat xenograft, SPARC expression was observed in the human glioma cells within the tumor mass, and in cells that invaded along vascular basement membranes and individually into the rat brain parenchyma, suggesting it may be an invasion-related gene. While it remains to be determined whether SPARC functionally contributes to tumor cell invasion, these data suggest that the early onset of increased SPARC expression, though complex, may serve as a signal indicative of neoplastic astrocytic transformation and reactive response to tumor-induced stress.

Dopaminergic innervation of substance P-containing striatal neurons by fetal nigral grafts: an ultrastructural double-labeling immunocytochemical study.

Evidence for survival and growth of fetal substantia nigra grafts in host striatum and partial reversal of behavioural and biochemical deficits in the host animal is well documented. Afferent synaptic connections arising from the graft and contacting host structures have also been reported; however, the properties of the neurons receiving this input is less clear. The purpose of this study was to determine if substance P-containing neostriatal neurons receive a dopaminergic input from nigral grafts. Fetal substantia nigra cell suspensions were stereotaxically implanted in the deafferented neostriatum of Wistar rats 2 weeks after a unilateral 6-hydroxydopamine (6-OHDA) lesion in the ipsilateral substantia nigra or medial forebrain bundle. The ultrastructural features of the graft-host synaptic interactions were analysed by employing an electron microscope immunocytochemical double-labeling technique. Tyrosine hydroxylase (TH) and substance P-immunoreactive structures were simultaneously demonstrated by means of the peroxidase-antiperoxidase method using two different chromogens with distinct reaction products easily differentiated at the light and electron microscope levels. TH-immunoreactive sites were first demonstrated using 3,3'-diaminobenzidine tetrahydrochloride (DAB); then substance P immunoreactivity was localized using benzidine dihydrochloride (BDHC). TH-immunoreactive terminals of axons originating from the graft made synaptic contacts with substance P-positive cell bodies and dendrites from the host. These results indicate that at least partial restoration of the normal nigrostriatal circuitry can be achieved following nigral grafts. The demonstration of specific synaptic input on host substance P neurons provides an anatomical basis for direct functional modulation of the deafferented host neostriatum by the nigral graft.

Parvalbumin-containing GABAergic neurons in the basal ganglia output system of the rat.

The output of the basal ganglia is directed through the entopeduncular nucleus (EPN) and the substantia nigra pars reticulata (SNR) and pars lateralis (SNL), which provide a gamma-aminobutyric acidergic (GABAergic) projection to various nuclei of the thalamus and brainstem. Although many neurons within the SNR and EPN have been described as modality specific, the morphological and neurochemical similarities preclude their precise identification. In the present study, the immunocytochemical localization of parvalbumin, a calcium-binding protein, is used in combination with axonal tracing to verify neuronal heterogeneity within the SNR, SNL, and EPN. The results reveal that the majority of neurons in all three centers contain parvalbumin. The parvalbumin-containing neurons are distributed in the caudal two-thirds of the EPN, the rostral part of the SNL, and the lateral two-thirds of the entire rostrocaudal extent of the SNR, the areas involved in sensorimotor function of the basal ganglia. Moreover, the nigrothalamic, nigrocollicular, and EPN-thalamic neurons possess parvalbumin immunoreactivity, whereas the EPN-habenular neurons are devoid of parvalbumin immunoreactivity. The results indicate a neurochemical heterogeneity within the GABAergic output neurons of the basal ganglia and suggest that the parvalbumin-containing neurons of the SNR, SNL, and EPN are the tonically active output neurons that form a major link in the disinhibitory neuronal circuit of the basal ganglia, especially that concerned with sensorimotor function.

Compartmental origin of the striato-entopeduncular projection in the rat.

The mammalian neostriatum is divisible into neurochemically and cytoarchitectonically distinct striosome and matrix compartments. This compartmentalization is respected by many afferent and efferent projections of the striatum. The distribution of distinct types of neuroactive substances and receptors and the unique connections of the striosome and matrix suggest a functional segregation between these compartments. The present study examines the organization of efferent projections from each of the striatal compartments to the entopeduncular nucleus (EPN), a major output center of the basal ganglia. The fluorescent retrograde tracer fluorogold, or rhodamine-conjugated dextran, was injected into the lateral habenula or the ventrolateral nucleus of the thalamus of adult Wistar rats to identify the topographical organization of EPN-habenular and EPN-thalamic neurons. Fluorogold was then placed into the rostral or caudal parts of the EPN, identified from the previous experiment as areas containing predominantly EPN-habenular or EPN-thalamic neurons, respectively. Sections containing retrogradely labeled neurons in the neostriatum were simultaneously immunolabeled for calbindin-D28kDa, a calcium-binding protein found exclusively in the projection neurons of the matrix. The results indicate that the striatal projection to the EPN-habenular and EPN-thalamic parts of the EPN originates from striosome and matrix neurons, respectively. The duality of striatal outflow involving the EPN suggests a mechanism whereby the striosome is integrated into subcortical pathways that modulate the activity of the basal ganglia via the ascending serotoninergic projection from the dorsal raphe nucleus, whereas the matrix is involved in a loop that includes the thalamus and the cerebral cortex.

Neurochemical compartmentalization of the globus pallidus in the rat: an immunocytochemical study of calcium-binding proteins.

The globus pallidus external segment forms a major target center of the mammalian striatum which is characterized by neurochemically distinct compartments. The present study was undertaken to determine if a corresponding compartmentalization exists within the globus pallidus external segment in the rat. Immunocytochemical examination of the calcium-binding proteins parvalbumin and calbindin D28kDa, which are present in neurons of the striatal matrix compartment, was employed. The results indicate three neurochemically distinct compartments within the globus pallidus external segment: 1) an area in the medial aspect of the entire length of the globus pallidus that contains dense immunoreactivity for calbindin D28kDa; 2) a narrow rim at the striatopallidal junction in the rostral two-thirds of the globus palidus that contains calbindin D28kDa immunoreactivity designated as the "border zone" of the globus pallidus; and 3) an area between these two zones showing very poor immunoreactivity for calbindin D28kDa but containing parvalbumin immunoreactive neurons. The calbindin D28kDa immunoreactive border zone corresponds to the area of the globus pallidus where striatal inputs converge extensively, whereas the rest of the nucleus is involved in segregated, topographically organized pathways. Parvalbumin-containing neurons are involved in the propagation of striatal output related to striosomal and sensorimotor aspects of basal ganglia function. The present results also indicate that calbindin D28kDa immunoreactivity is completely absent from striosomal neurons and is therefore a useful marker for striatal compartments.

Surgical treatment of intractable epilepsy attributable to multiple sclerosis.

Surgically removing a focus of epileptogenicity attributable to a multiple sclerosis (MS) plaque has not previously been considered a treatment option. Medically intractable partial epilepsy due to a chronically situated MS plaque is uncommon because most cases are self-limiting or managed with antiepileptic medication. We report a case of partial epilepsy resulting from such a plaque situated at the gray-white interface in the anterior parahippocampal gyrus. A favorable outcome was achieved by resection of the epileptogenic area.

Cardiovascular afferent and fastigial nucleus inputs to paramedian reticulospinal neurons.

In chloralose anesthetized, paralyzed and artificially ventilated cats, the region of the paramedian reticular nucleus (PRN) was systematically explored for single units antidromically activated by electrical stimulation of histologically verified sites in the intermediate gray region of the upper thoracic cord (T2). These antidromically identified units were then tested for their orthodromic responses to electrical stimulation of ipsilateral carotid sinus nerve (CSN) and of pressor sites in the contralateral fastigial nucleus (FN). Sixty-two histologically verified single units, located predominantly in the caudal half of the ventral PRN, were antidromically activated with latencies corresponding to a mean conduction velocity of 36.4 +/- 2.1 m/s. Of these units 25 (40%) were excited orthodromically by stimulation of the CSN and/or FN: 5 to stimulation of the CSN only (mean latency, 18.3 +/- 9.9 ms), 6 to stimulation of the FN only (mean latency, 7 +/- 1.7 ms), and 14 to stimulation of both the CSN and FN (mean latencies, 12.3 +/- 2.9 ms and 8.4 +/- 1 ms, respectively). These data provide electrophysiological evidence for the existence of PRN reticulo-spinal neurons that integrate and relay cardiovascular afferent information from the CSN and FN to spinal autonomic neurons.

The pallidostriatal projection in the rat: a recurrent inhibitory loop?

The pallidostriatal projection in the rat was investigated employing the PHA-L tracing technique. Following inotophoretic injections into the lateral aspect of the globus pallidus external segment, the ipsilateral striatum showed patches of dense anterograde labeling separated by areas containing sparse anterograde labeling and isolated retrogradely labeled neurons. The densely labeled patches did not correspond to any known compartments of the striatum. The retrogradely labeled neurons consistently showed similar distribution and morphological features reminiscent of striatal type II projection neurons. As all projection neurons of the striatum and all pallidal neurons are GABAergic, the complementary pattern of anterogradely and retrogradely labeled profiles from the globus pallidus suggest a possible mechanism whereby a horizontal inhibition may be exerted on groups of striatal neurons via the striato-pallido-striatal pathway.

GABAergic synaptic interactions in the substantia nigra.

The substantia nigra receives a strong GABAergic input from the ipsilateral striatum and globus pallidus. Nigral GABAergic synaptic interactions have been described in the pars compacta (SNC) and pars reticulata (SNR) but not in the pars lateralis (SNL). The SNR and particularly the SNL are the nodal points of the GABAergic nigrotectal pathway. The present study analyzes the synaptic connections of GABAergic and dopaminergic neurons in each of the divisions of the substantia nigra by employing a double-labeling immunocytochemical technique at the light and electron microscope levels. Glutamic acid decarboxylase (GAD)-containing terminals make symmetrical synaptic contacts with dopaminergic neurons in the SNC and SNR. Neurons that contain GAD also receive a GABAergic input in the SNR and SNL. The proportion of GAD-GAD contacts appears to be highest in the SNL where virtually all GAD-positive terminals are found to be in synaptic contact with or apposed to GAD positive profiles. This study demonstrates a strong GABAergic input onto nigral dopaminergic neurons and GABAergic neurons in the SNR and SNL. This GABAergic influence which is ostensibly striatal or pallidal in origin is particularly prominent in relation to the SNL-mediated nigro-collicular pathway.

Biotinylated dextran: a versatile anterograde and retrograde neuronal tracer.

A powerful and versatile axonal tracing method using biotinylated dextran, a novel analogue of biotin, is described. Pressure injection of varying volumes of 5% biotinylated dextran into various parts of the brain and spinal cord resulted in Golgi-like retrograde labeling and PHA-L-like anterograde labeling. The tracer filled the finest processes, revealing terminal axonal ramifications, distal dendrites and dendritic spines and excrescences. Extensive anterograde and retrograde labeling occurred in all pathways studied and in animals of all ages. Labeling appeared as early as 48 h and remained unchanged up to 14 days following injection. Biotinylated dextran can be detected easily with any avidin-conjugated marker for light and electron microscopic study. The resultant labeling can be combined readily with other morphological methods, such as tract tracing and/or immunocytochemical demonstration of endogenous substances. Biotinylated dextran is thus an efficient anterograde and retrograde tracer that can be combined with other neuroanatomical techniques to study details of synaptic interaction at all levels of dendritic organization.

Cerebellar, medullary and spinal afferent connections of the paramedian reticular nucleus in the cat.

The topographic organization of afferent projections from the deep cerebellar nuclei, medulla oblongata and spinal cord to the paramedian reticular nucleus (PRN) of the cat was studied using the horseradish peroxidase (HRP) method of retrograde labelling. Discrete placements of HRP within each of the dorsal (dPRN) and ventral (vPRN) regions of the PRN showed some segregation of input. The deep cerebellar nuclei project in a predominantly contralateral fashion upon the PRN. A small but significant ipsilateral fastigial afferent component is also present. The fastigial and dentate nuclei contribute the majority of fibers to the dPRN whereas the interposed nucleus provides very little. The vPRN receives a relatively uniform input from all 3 cerebellar nuclei. Both lateral vestibular nuclei contribute the majority of fibers from the vestibular nuclear complex largely from their dorsal division. Additional input arises from bilateral medial and inferior vestibular nuclei. The vPRN receives relatively more fibers from the inferior vestibular nuclei than does the dPRN while inputs from the medial vestibular nuclei are comparably sparse. The PRN receives bilateral projections from the nucleus intercalatus (of Staderini). A significant projection to the contralateral PRN occurs from the ventrolateral subnucleus of the solitary complex and its immediate vicinity. Additional sources of medullary afferent input include the lateral, gigantocellular and magnocellular tegmental fields, the contralateral PRN and the raphe nuclei. Sites of origin of spinal afferents to the dPRN are bilaterally distributed mainly within Rexed's laminae VII and VIII of the cervical cord whereas those to the vPRN are confined largely to the medial portion of the contralateral lamina VI in the C1 segment. A few labelled cells are found in the thoracolumbar cord with those to the vPRN being more caudal. These data provide the neuroanatomical substrate for a better understanding of the functional role of the PRN in mediating cardiovascular responses appropriate to postural changes.

Effects of injection mechanics, pH of infusate and 6-hydroxydopamine on cerebromicrovascular permeability in rats.

The effects of altering the rate, manner and vehicle used for intracerebral injection upon microvascular permeability were studied in Sprague-Dawley rats employing horseradish peroxidase histochemistry. The volume of vehicle delivered and the site of intracerebral injection were kept constant. In comparison to continuous infusion, vascular permeability was significantly greater following manual (intermittent) injections; however, no differences were found when the infusion rate was decreased 10-fold. Use of a buffered vehicle (Hanks' balanced salt solution) with pH adjusted to 7.4, in contrast to the more commonly used non-buffered vehicle (saline-ascorbate), resulted in significant reductions in permeability. The apparent influence of the agent 6-hydroxydopamine (6-OHDA) on changes in vascular permeability was found to vary depending on the type and pH of the vehicle used for injection. Significantly greater permeability resulted with saline-ascorbate (pH 3.1) as the vehicle when compared to Hanks' balanced salt solution (pH 7.4). Changes in vascular permeability can therefore be produced by varying mechanical and vehicular factors which, in the case of 6-OHDA, far outweigh previously reported permeability changes specifically attributed to this neurotoxin.

Collateral branching in axonal projections to spinal cord from paramedian reticular nucleus neurons.

Experiments were done in cats to identify neurons in the paramedian reticular nucleus (PRN) sending collateral axons to the region of the intermediolateral nucleus (IML) at different levels of the thoracic cord by using lectin-conjugated horseradish peroxidase (HRP) and double-labeling fluorochrome histochemistry to retrogradely label PRN neurons. Injections of Fast blue (FB) into the spinal cord at the T2 level centered in the region of the IML were coupled with injections of Nuclear yellow (NY) into the ipsilateral cord at either the T4 or T7 levels centered in the region of the IML. Neurons in the PRN retrogradely labeled after diffusion of HRP into the region of the IML at the T2 level were observed throughout the rostrocaudal extent of the ventral PRN. In addition, a few labeled neurons were noted in the ventral portion of the dorsal PRN. About 40% of the neurons in the PRN which were labeled with FB after an injection at the T2 level were also labeled with NY injected into the cord in further caudal segments. These data suggest that the PRN may exert its influence on the cardiovascular system partly through collateral axonal branches to widely separated populations of sympathetic preganglionic neurons in different spinal segmental levels.

Vestibular nucleus inputs to paramedian reticulospinal neurons in the cat.

Experiments were done in chloralose-anesthetized cats to identify single units in the paramedian reticular nucleus (PRN) that responded to stimulation of pressor sites in the vestibular nucleus complex (VNC) and that projected directly to the intermediate gray (IG) region of the upper thoracic cord. Forty-seven units responded orthodromically to stimulation of the ipsilateral VNC with a mean latency of 6.3 +/- 0.6 ms: 44 were excited and 3 were inhibited. Of these 47 units, 29 (62%) also were antidromically activated by stimulation of the ipsilateral IG at the level of T2. These data provide electrophysiological evidence for the existence of neurons in PRN that receive VNC inputs and project directly to spinal autonomic areas, and suggest that this pathway may be involved in mediating vestibulosympathetic reflex responses associated with postural adjustments.

Axonal branching in the projections from the paramedian reticular nucleus to the cerebellar cortex.

Injections of fluorescent tracers into cat cerebellar cortex gave evidence of collateral axonal branching of neurons situated in the paramedian reticular nucleus. These branched reticulocerebellar projections were distributed to opposing sides of the cerebellum, in particular the anterior lobe and the ansiform lobule. No topographical organization was observed in the PRN. Less than 30% of ipsilaterally projecting reticulocerebellar fibers had contralaterally directed collateral branches. These results are in keeping with a bilateral fastigial projection to the PRN forming a feedback loop circuit through which orthostatic reflexes may be mediated.

Size estimation and magnification error in radiographic imaging: implications for classification of arteriovenous malformations.

PURPOSE: To assess magnification error in digital subtraction angiography as it pertains to arteriovenous malformation (AVM) size. METHODS: A rectangular grid phantom with equally spaced markers mounted in a stereotactic frame was imaged with digital angiographic equipment. The location and orientation of the grid was altered relative to the central plane of the phantom. Both linear and area measurements were made according to the perceived location of phantom markers using a standard catheter calibration technique and compared with stereotactically derived estimates. Finally, a single case example of an angiographically imaged rolandic AVM was used to compare linear dimensions obtained with both described techniques. RESULTS: The determination of location and size with standard angiographic imaging is subject to error because of the divergent geometry of the incident x-ray beam. The resulting nonconstant geometric magnification causes errors in linear measurements of 10% to 13% at depths of 7 cm from the calibration plane. Errors in area measurements at the same position increase by 20% to 25%. Measurements of maximum diameter or cross-sectional area may have an additional error when nonspherical objects are inclined to the viewing direction (40% at 45 degrees inclination). These errors are reduced to less than 1 mm using the stereotactic technique. Some commercial angiographic systems have internal software to enable a spatial calibration based on known distances in the image or on the diameter of a catheter. The catheter technique was accurate in the calibration direction (perpendicular to the catheter axis) but had a 12% error in the direction parallel to the catheter because of a nonunity aspect ratio in the video system. Measurement of the dimensions of a rolandic AVM using the catheter calibration technique had an error that ranged from -3% to +26% (standard error, 20%) with respect to the stereotactic technique. CONCLUSIONS: Numerous nonstereotactic referential systems for determining linear distances are inherently erroneous by varying degrees compared with the stereotactic technique. Area and volume determinations naturally increase this error further. To the extent that no standardized method for determining linear distances exists, significant variations in estimation of AVM size result. Classification schemes for AVMs have been hampered by this technical error.

Substance P synaptic interactions with GABAergic and dopaminergic neurons in rat substantia nigra: an ultrastructural double-labeling immunocytochemical study.

The distribution of substance P (SP), tyrosine hydroxylase (TH), and glutamic acid decarboxylase (GAD) immunoreactivity in the substantia nigra of the rat was studied by means of an ultrastructural double-labeling immunocytochemical method. Direct synaptic contact between SP-immunoreactive terminals and GAD-positive nigral neurons was more often observed in the pars lateralis than the pars reticularis and was rarely observed in the pars compacta. Substance P-positive terminals also formed synapses with cell bodies and dendrites of TH-positive, dopaminergic neurons in the pars compacta and pars reticulata. Multiple SP-immunoreactive terminals were often observed with symmetrical and, less frequently, asymmetrical synapses on individual TH-containing dendrites. Evidence of SP-containing terminals contacting both GABAergic and dopaminergic neurons in the substantia nigra suggests a direct excitatory action upon nigral projection neurons.