Does astrocyte gap junction protein expression differ during development in absence epileptic rats?

Intercellular communication via gap junctions (GJs) has a wide variety of complex and essential functions in the CNS. In the present developmental study, we aimed to quantify the number of astrocytic GJs protein connexin 30 (Cx30) of genetic model of absence epilepsy rats from Strasbourg (GAERS) at postnatal P10, P30, and P60 days in the epileptic focal areas involved in the cortico-thalamic circuit. We compared the results with Wistar rats using immunohistochemistry and western blotting. The number of Cx30 immunopositive astrocytes per unit area were quantified for the somatosensory cortex (SSCx), ventrobasal (VB), and lateral geniculate (LGN) thalamic nuclei of the two strains and Cx30 western blot was applied to the tissue samples from the same regions. Both immunohistochemical and western blot results revealed the presence of Cx30 in all regions studied at P10 in both Wistar and GAERS animals. The SSCx, VB, and LGN of Wistar animals showed progressive increase in the number of Cx30 immunopositive labeled astrocytes from P10 to P30 and reached a peak at P30; then a significant decline was observed from P30 to P60 for the SSCx and VB. However, in GAERS Cx30 immunopositive labeled astrocytes showed a progressive increase from P10 to P60 for all brain regions studied. The immunohistochemical data highly corresponded with western blotting results. We conclude that the developmental disproportional expression of Cx30 in the epileptic focal areas in GAERS may be related to the onset of absence seizures or may be related to the neurogenesis of absence epilepsy.

Anatomic variations of the human falx cerebelli and its association with occipital venous sinuses.

PURPOSE: Human falx cerebelli is an important anatomical structure in regard to its relations with venous structures during infratentorial approach to reach cerebellar tumors, vascular malformations, traumatic hemorrhage and Chiari malformations. The present study aim to describe the different types of variations of the falx cerebelli, its morphological features and its association with occipital venous sinuses. METHOD: In this study 49 dura mater was obtained from the Institution of Forensic Medicine. The length, width and the depth of the falx cerebelli were measured using a digital compass. The data obtained were statistically analyzed in relation to age and gender. The relations of the falx cerebelli with the occipital sinus was documented. Histological sections from the falx cerebelli were stained with Hematoxylin Eosin to evaluate the fine structure. RESULTS: Among the 49 falx cerebelli examined 36 (73.5%) were classified as normal. The average length, width and depth of the normal falx cerebelli was 3.7, 1.0 and 0.4 cm respectively. Of the 49 falx cerebelli in 1 (2%) case it was absent, in 5 cases (10.2%) duplicate, in 5 cases (10.2%) triplicate, in 1 (2%) case quadruplets and in 1 case (2%) it was five-folded. The proximal and the distal attachments of the falx cerebelli showed 3 types of variations; both attachments triangular, the proximal attachments triangular and the distal ramified and distal attachments triangular and the proximal attachments ramified. The drainage of the occipital sinus of falx cerebelli with variations were evaluated. The increased number of falx cerebelli highly corresponded with the increased number of occipital sinus. CONCLUSIONS: The dural-venous variation in the posterior cranial fossa can be problematic in various diagnostic and operative procedures of this region. Neurosurgeons should be aware of such variations, as these could be potential sources of haemorrhage during the midline suboccipital and infratentorial approaches.

The Cerebello-Hypothalamic and Hypothalamo-Cerebellar Pathways via Superior and Middle Cerebellar Peduncle in the Rat.

The connections between the cerebellum and the hypothalamus have been well documented. However, the specific cerebellar peduncle through which the hypothalamo-cerebellar and cerebello-hypothalamic connections pass has not been demonstrated. The present study aims to define the specific cerebellar peduncle through which connects the cerebellum to specific hypothalamic nuclei. Seventeen male albino rats received 20-50-nl pressure injections of either Fluoro-Gold (FG) or biotinylated dextran amine (BDA) tracer into the superior (SCP), middle (MCP), and inferior (ICP) cerebellar peduncle. Following 7-10 days of survival period, the animals were processed according to the appropriate protocol for the two tracers used. Labeled cells and axons were documented using light or fluorescence microscopy. The present study showed connections between the hypothalamus and the cerebellum via both the SCP and the MCP but not the ICP. The hypothalamo-cerebellar connections via the SCP were from the lateral, dorsomedial, paraventricular, and posterior hypothalamic nuclei, and cerebello-hypothalamic connections were to the preoptic and lateral hypothalamic nuclei. The hypothalamo-cerebellar connections via the MCP were from the lateral, dorsomedial, ventromedial, and mammillary hypothalamic nuclei; and cerebello-hypothalamic connections were to the posterior, arcuate, and ventromedial hypothalamic nuclei. The hypothlamo-cerebellar connections were denser compared to the cerebello-hypothlamic connections via both the SCP and the MCP. The connection between the cerebellum and the hypothalamus was more prominent via the SCP than MCP. Both the hypothlamo-cerebellar and cerebello-hypothalamic connections were bilateral, with ipsilateral preponderance. Reciprocal connections were with the lateral hypothalamic nucleus via the SCP and the ventromedial nucleus via the MCP were observed. Cerebellum takes part in the higher order brain functions via its extensive connections. The knowledge of hypothalamo-cerebellar and cerebello-hypothalamic connections conveyed within the SCP and MCP can be important for the lesions involving the MCP and SCP. These connections can also change the conceptual architecture of the cerebellar circuitry and deepen current understanding.

Cortical, subcortical and brain stem connections of the cerebellum via the superior and middle cerebellar peduncle in the rat.

The role of cerebellum in coordination of somatic motor activity has been studied in detailed in various species. However, experimental and clinical studies have shown the involvement of the cerebellum with various visceral and cognitive functions via its vast connections with the central nervous system. The present study aims to define the cortical and subcortical and brain stem connections of the cerebellum via the superior (SCP) and middle (MCP) cerebellar peduncle using biotinylated dextran amine (BDA) and Fluoro-Gold (FG) tracer in Wistar albino rats. 14 male albino rats received 20-50-nl pressure injections of either FG or BDA tracer into the SCP and MCP. Following 7-10 days of survival period, the animals were processed according to the related protocol for two tracers. Labelled cells and axons were documented using light and fluorescence microscope. The SCP connects cerebellum to the insular and infralimbic cortices whereas, MCP addition to the insular cortex, it also connects cerebellum to the rhinal, primary sensory, piriform and auditory cortices. Both SCP and MCP connected the cerebellum to the ventral, lateral, posterior and central, thalamic nuclei. Additionally, SCP also connects parafasicular thalamic nucleus to the cerebellum. The SCP connects cerebellum to basal ganglia (ventral pallidum and clastrum) and limbic structures (amygdaloidal nuclei and bed nucleus of stria terminalis), however, the MCP have no connections with basal ganglia or limbic structures. Both the SCP and MCP densely connects cerebellum to various brainstem structures. Attaining the knowledge of the connections of the SCP and MCP is important for the diagnosis of lesions in the MCP and SCP and would deepen current understanding of the neuronal circuit of various diseases or lesions involving the SCP and MCP.

The contributions to the human dorsal column tracts from the spinal cord laminae.

The dorsal column tracts (fasciculus gracilis and fasciculus cuneatus) are concerned with discriminative qualities of sensation. There are controversial descriptions related to the relations of dorsal column tracts with the dorsal horn laminae in text-books. The present study aims to define the laminae of the dorsal horn of the spinal cord that contribute fibers to the dorsal column tracts in the cervical, thoracic and lumbar spinal level. Series paraffin spinal cords sections of six formalin-embalmed adult human cadavers were evaluated. The present study shows that dorsal column tracts receive fiber contributions from laminae III and V and from Clarke's dorsal nucleus at varying spinal levels. At upper cervical levels (C1-C4) fiber contributions were from lamina V and few from lamina III, and at lower cervical levels (C5-C8) there were, in addition to these laminae, also contributions from the Clarke's dorsal nucleus. At upper thoracic levels (T1-T4) fiber contributions were from lamina V and few from Clarke's dorsal nucleus. At lower thoracic (T5-T12) and lumbar levels (L1-L5), in contrast, fiber contributions were only from Clarke's dorsal nucleus. The detailed knowledge of organization of the dorsal column tracts of the spinal cord may pave the way for future treatments of the spinal cord injuries.

Intradural communication between dorsal rootlets of spinal nerves: their clinical significance.

BACKGROUND: Anatomical and surgical textbooks give almost no attention to the intradural communications between dorsal rootlets of adjacent spinal nerves. These communications can be of significance in various neurosurgical procedures and clinical conditions of the region. METHODS: The spinal cord of six formaldehyde-fixed cadavers was dissected from C1-S5. The dorsal rootlets of the spinal nerves were exposed via a posterior approach and communications between adjacent spinal nerves were documented. RESULTS: The frequency of communication between adjacent dorsal rootlets of the spinal nerves showed variations among spinal levels. Thirty-eight dorsal rootlet communications were observed in six cadavers (12 sides) and 20 (52.6%) were at cervical levels, 14 (36.8%) at thoracic levels, and four (10.5%) at lumbar levels. The majority of communications were observed on the left side (65.8%). Communications were most frequently observed at cervical (C4-C5, C5-C6) and upper thoracic (T1-T2) levels and seen least frequently at lower thoracic and lumbar levels. No communications were observed at sacral levels. Five types of communication were observed: I. oblique ascending, II. oblique descending III. short Y, IV. long Y and V shaped. None of the communication extended beyond one segment at any spinal level. The occurrence of such dorsal rootlet communications ranged from 3 to 7 for each cadaver and the mean was 4.8 ± 1.3. Histological sections from various levels of the dorsal rootlet communications showed that all consisted of myelinated fibers of varying diameters. CONCLUSIONS: Such communications may lead to misinterpretation of the pathology on the basis of clinical signs and symptoms and also should be considered in rhizotomy.

The Supraorbital Keyhole Approach.

AIM: The major aim of the present anatomical study was to demonstrate the anatomical structures that can be visualized using the supraorbital keyhole approach, both endoscopically and microscopically, from an eyebrow incision to intracranial structures. Furthermore, it defines an optimal craniotomy for surgery. METHODS: Fine dissection was performed on each side of 5 formalin-fixed adult cadavers according to the surgical procedures of the supraorbital keyhole approach, and each step was documented both endoscopically and microscopically. Furthermore, the distance between the superior temporal line and the supraorbital notch/foramen was measured from the 10 total sides of the 5 cadavers and from the 118 sides of the 59 autopsies. RESULTS: Tumors and aneurysms of the anterior cranial fossa can be visualized during the supraorbital keyhole approach. The average distance between the superior temporal line and the supraorbital notch/foramen was measured. The distance obtained from the autopsies on the 25 females was 31.56 ±â€Š4.03  mm on the right side and 31.04 ±â€Š5.40  mm on the left side. The average distance obtained from the autopsies on the 34 males was 34.00 ±â€Š4.59  mm on the right side and 33.59 ±â€Š5.41  mm on the left side. There was no statistically significant difference between right and left in the female and male autopsies or between sexes. CONCLUSIONS: This anatomical study showed that structures in the anterior and middle cranial fossa can be reached via the supraorbital keyhole craniotomy approach with minimal brain retraction and adequate exposure and with minimal craniotomy size.

Anatomy of the spinal dorsal root entry zone: its clinical significance.

BACKGROUND: The posterolateral sulcus (PLS) is an important surgical landmark, especially for DREZ (dorsal root entry zone) operations. METHODS: The present study aimed to show the variations of the PLS using human spinal cord histological sections and report the variability in the number of dorsal rootlets of the spinal nerves in each the spinal cord segment. Further, measure the height and width of the dorsal horn on histological sections for cervical, thoracic, and lumbar levels. RESULTS: The results of the present study showed various patterns of PLS 1.clearly present PLS, 2. short PLS, 3. absent PLS or 4. irregular PLS. Height and width measurements of the dorsal horn showed that the average width was greatest at lower cervical (0.48 ± 0.04 mm) and least at lower thoracic levels (0.41 ± 0.04 mm), whereas the average height was greatest at upper cervical (3.0 ± 0.06 mm) and smallest at lower lumbar levels (1.8 ± 0.08 mm). The average number of rootlets varied considerably, at cervical level it was 7.6 ± 1.4 mm, at thoracic 6.6 ± 0.8 mm and at lumbar 6.1 ± 0.4 mm. CONCLUSIONS: The detailed anatomy of the variations of the PLS and the average number of rootlets at each spinal level can increase the success of regional surgery. Further, fine measurements on histological sections can give detailed knowledge on the size necessary for lesioning in DREZ operations.

Motor afferents from the cerebellum, zona incerta and substantia nigra to the mediodorsal thalamic nucleus in the rat.

The mediodorsal (MD) thalamic nucleus provides information from subcortical structures to the prefrontal cortex. The human MD thalamic nucleus has been implicated in a great variety of different clinical conditions and normal functions ranging from schizophrenia, Parkinsonism and epilepsy to many cognitive functions. In the rat the MD thalamic nucleus is divided into three cytoarchitectonic sectors whereas in the primates it is divided into two; medial one-third (magnocellular) and lateral two-thirds further the lateral sector is divided into pars parvocellularis pars multiformis, pars fasciculosa and pars caudalis. In this study we used a retrograde tracer, fluoro-gold (FG) to evaluate some of the afferents reaching the lateral sector of the MD (MDl) thalamic nucleus. The results of the present study have shown that MDl receives afferent connections from the lateral cerebellar nucleus (dentate nucleus), substantia nigra pars reticulata (SNR) and zona incerta (ZI). Subsequent to FG injections into the MDl, labeled cells were observed mainly bilaterally but were sparser on the contralateral side than ipsilaterally from each of the three structures listed. All three afferents showed a topographical organization. The labeled neurons were localized at the dorsomedial aspect of the lateral cerebellar nucleus, the dorsoventral aspect of the SNR and in the dorsal sector of the ZI. The lateral cerebellar nucleus reached the MDl via the superior cerebellar peduncle. No other deep cerebellar nuclei showed labeled cells. There were no labeled cells in the substantia nigra pars compacta (SNC). Although the three regions identified here are recognized as having motor functions, the connections to MD suggest that their outputs also play a role in cognitive or other higher cortical functions.

Extracranial transport of brain lymphatics via cranial nerve in human.

Extracranial waste transport from the brain interstitial fluid to the deep cervical lymph node (dCLN) is not extensively understood. The present study aims to show the cranial nerves that have a role in the transport of brain lymphatics vessels (LVs), their localization, diameter, and number using podoplanin (PDPN) and CD31 immunohistochemistry (IHC) and Western blotting. Cranial nerve samples from 6 human cases (3 cadavers, and 3 autopsies) were evaluated for IHC and 3 autopsies for Western blotting. The IHC staining showed LVs along the optic, olfactory, oculomotor, trigeminal, facial, glossopharyngeal, accessory, and vagus nerves. However, no LVs present along the trochlear, abducens, vestibulocochlear, and hypoglossal nerves. The LVs were predominantly localized at the endoneurium of the cranial nerve that has motor components, and LVs in the cranial nerves that had sensory components were present in all 3 layers. The number of LVs accompanying the olfactory, optic, and trigeminal nerves was classified as numerous; oculomotor, glossopharyngeal, vagus, and accessory was moderate; and facial nerves was few. The largest diameter of LVs was in the epineurium and the smallest one was in the endoneurium. The majority of Western blotting results correlated with the IHC. The present findings suggest that specific cranial nerves with variable quantities provide a pathway for the transport of wastes from the brain to dCLN. Thus, the knowledge of the transport of brain lymphatics along cranial nerves may help understand the pathophysiology of various neurological diseases.

Comparing GABAergic cell populations in the thalamic reticular nucleus of normal and genetic absence epilepsy rats from Strasbourg (GAERS).

The GABAergic neurons of the thalamic reticular nucleus (TRN) play a critical role in the generation and control of spike-and-wave discharges (SWDs) in absence epilepsy. We have used the disector method to count the GABA+ve and GABA-ve neurons in the intermediate TRN sector of genetic absence epilepsy rats from Strasbourg (GAERS) and of Wistar rats during postnatal (P) development at P10, P20, P30, and P60 days. The same part of TRN was removed from each animal, the GABAergic neurons were labelled using light-microscopical GABA immunohistochemistry and the data were statistically analysed. Both the GAERS and Wistar animals showed an increase in the density of GABA+ve and GABA-ve cells from P10 to P20. From P20 to P60, Wistar animals showed no significant differences for either cell type, but in the GAERS a progressive decrease from P20 to P60 was observed in both GABA+ve and GABA-ve cells. The decrease of the GABA-ve cells was more pronounced than that of the GABA+ve cells. There were no significant differences between cell sizes for GAERS and Wistar rats at any developmental age. The lower density GABA+ve and GABA-ve neurons at P30 and P60 of GAERS compared to Wistar animals may contribute to the generation of SWDs in absence epilepsy.

Falcine venous plexus within the falx cerebri: anatomical and scanning electron microscopic findings and clinical significance.

BACKGROUND: Only limited attention has been paid to the anatomy and clinical importance of the falcine venous plexus. The aim of this study was to evaluate the falcine venous plexus anatomically using scanning electron microscopy (SEM), and to provide guidance for neurosurgical approaches. METHODS: Latex or ink was injected into the superior and inferior sagittal sinus. The falcine venous plexus lying within the connective tissue of the falx cerebri was observed by dividing the falx into thirds (anterior, middle and posterior). Further, the SEM appearance of the falcine venous plexus was evaluated. RESULTS: The anterior third of the falx cerebri consisted of small diameter falcine venous vessels. These vessels were localized close to either the superior or inferior sagittal sinus, and none extended as far as mid-falx cerebri levels in any of the 16 cases. They communicated with either superior or inferior sagittal sinuses, but not with both of these sinuses. In the middle third of the falx cerebri, the majority of the vessels of the falcine venous plexus had larger diameter compared to those of the anterior third. These vessels extended the length of the falx cerebri levels. They communicated with both superior and inferior sagittal sinuses. In the posterior third of the falx cerebri, the vessels of the falcine venous plexuses had the largest diameter and were located at the junction of the inferior sagittal sinus and the straight sinus. They were localized at the lower two-thirds of the falx cerebri. In all cases, the dense venous networks communicated with the inferior sagittal sinus but not with the superior sagittal sinus. The falcine venous plexus observed in the posterior third of the falx cerebri was denser than in the anterior and middle portions. The SEM revealed small vessels whose diameter ranged between 42 and 138 µm. The vessels of the falcine venous plexus in the anterior third had a mean diameter of 0.42 ± 0.1 mm, in the middle third a mean diameter of 0.87 ± 0.17 mm, and in the posterior third, 1.38 ± 0.21 mm. CONCLUSION: The falcine venous plexus is a network of venous channels that exists within the connective tissue of the falx; the sizes and patterns of communication of these structures showed regional differences. Neurosurgeons should be aware of the regional differences when making an incision or puncturing the falx during a surgical approach.

Regional connections of the mediodorsal thalamic nucleus in the rat.

Thalamic nuclei are classified as first- and higher-order relays. The first-order relays receive their driving afferents from ascending pathways and transmit messages to cortex that cortex has not seen before. The higher-order relays receive driver messages from layer-5 cortical cells for transmission from one cortical area to another. The present study used the retrograde tracer, fluoro-gold, to define the afferents to the three regions of the mediodorsal thalamic nucleus, to distinguish which parts contain first- or higher-order relays. The results show that the main inputs to the medial region of the nucleus come from olfactory and visceral structures, those to the central region come from limbic structures and those to the lateral region come from motor centers of the central nervous system. The medial and central regions receive both modulatory (layer 6) and driver (layer 5) afferent inputs from the orbitofrontal and medial frontal areas of the prefrontal cortex whereas the lateral region receives no layer-5 inputs from its cortical connections. Further, the inhibitory modulation of the mediodorsal thalamic nucleus shows regional differences. The medial region receives inhibitory afferents from the striatum (globus pallidus, caudate-putamen), the lateral region from the substantia nigra pars reticulata and the zona incerta, and all segments of the mediodorsal thalamic nucleus receive inhibitory afferents from the thalamic reticular nucleus. The results of the present study show that each region of the mediodorsal thalamic nucleus has distinct afferent connections allowing each region of mediodorsal thalamic nucleus to be considered relatively independent subnuclei that may subserve independent functions.

The brainstem connections of the supplementary motor area and its relations to the corticospinal tract: Experimental rat and human 3-tesla tractography study.

Although the supplementary motor area (SMA) is a large region on the medial surface of the frontal lobe of the brain, little is known about its function. The current study uses 3-tesla high-resolution diffusion tensor tractography (DTI) in healthy individuals and biotinylated dextran amine (BDA) and fluoro-gold (FG) tracer in rats to demonstrate the afferent and efferent connections of the SMA with brainstem structures. It also aims to clarify how SMA fibers relate to the corticospinal tract (CST). The BDA (n = 6) and FG (n = 8) tracers were pressure-injected into the SMA of 14 Wistar albino rats. Light and fluorescence microscopy was used to capture images of the FG and BDA-labeled cells and axons. High-resolution 3-tesla DTI data were acquired from the Human Connectome Project database. Tracts between the SMA and brainstem structures were analyzed using diffusion spectrum imaging (DSI) studio software. The FG injections into the SMA showed afferent projections from mesencephalic (periaqueductal gray matter, substantia nigra pars reticulata, ventral tegmental area, inferior colliculus, mesencephalic reticular, tegmental, and raphe nuclei), pontine (locus coeruleus, pontine reticular and vestibular nuclei), and medullary (area postrema, parabrachial, and medullary reticular nuclei) structures. The anterograde tracer BDA injections into the SMA showed efferent connections with mesencephalic (periaqueductal gray, substantia nigra pars compacta, dorsal raphe, trigeminal motor mesencephalic, and mesencephalic reticular nuclei), pontine (locus coeruleus, nucleus of the lateral lemniscus, vestibular, cochlear, and pontine reticular nuclei), and medullary (area postrema, medullary reticular, olivary, and parabrachial nuclei) structures. The SMA had efferent but no afferent connections with the cerebellar nuclei. The DTI results in healthy human subjects highly corresponded with the experimental results. Further, the DTI results showed a distinct bundle that descended to spinal levels closely related to the CST. Understanding SMA's afferent and efferent connections will enrich our knowledge of its contribution to various brainstem networks and may provide new perspectives for understanding its motor and non-motor functions.

Do the quantitative relationships of synaptic junctions and terminals in the thalamus of genetic absence epilepsy rats from Strasbourg (GAERS) differ from those in normal control Wistar rats.

Abnormal functional properties of the thalamocortical connections were reported in the absence of epilepsy. The present study compares the ratios of terminals ('RL'-round vesicles, large terminals, 'RS'-round vesicles, small terminals and 'F'-flattened vesicles) and synapse in three first-order (ventrobasal, lateral geniculate and anteroventral) and in three higher-order (posterior, lateral posterior and mediodorsal) thalamic nuclei of genetic absence epilepsy rats from Strasbourg (GAERS) with our earlier quantitative studies of normal Wistar rats to show whether quantitative differences were present in GAERS as compared to Wistar rat. Rats were perfused transcardially, the brains were removed and cut as 300 µm coronal sections. Parts of the six thalamic nuclei were removed for routine electron microscopy and GABA immunocytochemistry. Twenty photographs from each section at 20,000× magnification were taken, and the terminals were identified as RL, RS or F. (1) In normal Wistar rats (as in cats), the proportion of driver terminals (RL) and synapses is lower in higher-order than in first-order thalamic nuclei, but this difference is not present in GAERS animals. (2) The proportions of RS terminals and synapses for each thalamic nucleus showed no significant differences between GAERS and Wistar rats for any of the thalamic nuclei. (3) In GAERS, the proportion of inhibitory F terminals and synapses was significantly high in the VB and low in the LP thalamic nucleus. These abnormal ratios in the GAERS may be the cause of the spike-and-wave discharges of absence seizures or may represent a compensatory response of the thalamocortical circuitry to the absence seizures.

The denticulate ligament: anatomical properties, functional and clinical significance.

BACKGROUND: It is widely believed that the main function of denticulate ligaments (DLs) is to stabilize the spinal cord within the vertebral canal. The aim of this study was to assess the anatomical and histological structure of the DLs and to document any regional differences. METHODS: Five formalin-fixed adult cadavers were used. The DLs were exposed via the posterior approach, and detailed anatomy and histology of these structures were documented. RESULTS: The main findings were: (1) each DL is composed of a single narrow fibrous strip that extends from the craniovertebral junction to T12, and each also features 18-20 triangular extensions that attach to the dura at their apices; (2) the triangular extensions are smaller and more numerous at the cervical levels, and are larger and less numerous at the thoracic levels; (3) the apices of the extensions attach to the dura via fibrous bands at cervical levels (each band 3-5 mm long) and lower thoracic levels (21-26 mm long), whereas they attach directly to the dura at upper thoracic levels; (4) the narrow fibrous strip of the DL features longitudinally oriented collagen fibers, whereas the triangular extensions are composed of transverse and obliquely oriented collagen fibers. The collagen fibers are thicker and more abundant at the cervical than at the thoracic levels. CONCLUSION: DL histology and anatomy are strongly correlated with the function of this structure at different spinal levels. It is important to have accurate knowledge about DLs as these structures are relevant for clinical procedures that involve the spinal cord or craniovertebral junction.

Connections of the Dentate Nucleus with the Amygdala: Experimental Rat and Human 3-Tesla Tractography Study.

Background: The role of the cerebellum in motor function is well recognized. However, its role in higher nervous system activities such as cognition, emotion, endocrine, and autonomic activities is less known. The present study aims to show direct dento-amygdala projections using a biotinylated dextran amine (BDA) tracer in rats and 3-tesla (T) high-resolution diffusion tensor imaging (DTI)-based tractography in humans. Materials and Methods: The BDA tracer was pressure injected into the dentate nucleus of the cerebellum of Wistar albino rats. Labeled cells and axons were documented. High-resolution 3-T tractography data were obtained from the Human Connectome Project database. Dento-amygdala tracts were analyzed using diffusion spectrum imaging (DSI) Studio software. Results: The experimental study showed bilateral projections between the dentate nucleus and the central and basal nuclei and ipsilateral projections between lateral nuclei of the amygdala. The fibers from the dentate nucleus reached the amygdala through the superior cerebellar peduncle (SCP), and the contralateral fibers crossed in the decussation of SCP at the midbrain. The dento-amygdala results of the experimental study corresponded with the 3-T tractography findings on humans. Additionally, DTI findings showed that most of the dentate fibers passed through the hypothalamus before reaching the amygdala, and the amygdalae of the two sides are connected through the anterior commissure. Discussion: The 3-T DTI data of adult humans showed both direct dento-amygdala and indirect dento-hypothalamo-amygdala projections. Thus, this may indicate cerebellar contribution in modulation of emotional and autonomic functions. Furthermore, this can explain the emotional and cognitive deficits that occur in patients with cerebellar or SCP damage. Impact statement The present study showed direct dento-amygdala connections in the rat brain and human brain, which may provide evidence for cerebellar contribution in modulation of emotional and autonomic functions.

The Effects of Optogenetic Activation of Astrocytes on Spike-and-Wave Discharges in Genetic Absence Epileptic Rats.

Background: Absence seizures (petit mal seizures) are characterized by a brief loss of consciousness without loss of postural tone. The disease is diagnosed by an electroencephalogram (EEG) showing spike-wave discharges (SWD) caused by hypersynchronous thalamocortical (TC) oscillations. There has been an explosion of research highlighting the role of astrocytes in supporting and modulating neuronal activity. Despite established in vitro evidence, astrocytes' influence on the TC network remains to be elucidated in vivo in the absence epilepsy (AE). Purpose: In this study, we investigated the role of astrocytes in the generation and modulation of SWDs. We hypothesize that disturbances in astrocytes' function may affect the pathomechanism of AE. Methods: To direct the expression of channelrhodopsin-2 (ChR2) rAAV8-GFAP-ChR2(H134R)-EYFP or to control the effect of surgical intervention, AAV-CaMKIIa-EYFP was injected into the ventrobasal nucleus (VB) of the thalamus of 18 animals. After four weeks following the injection, rats were stimulated using blue light (~473 nm) and, simultaneously, the electrophysiological activity of the frontal cortical neurons was recorded for three consecutive days. The animals were then perfused, and the brain tissue was analyzed by confocal microscopy. Results: A significant increase in the duration of SWD without affecting the number of SWD in genetic absence epileptic rats from Strasbourg (GAERS) compared to control injections was observed. The duration of the SWD was increased from 12.50 ± 4.41 s to 17.44 ± 6.07 following optogenetic stimulation in GAERS. The excitation of the astrocytes in Wistar Albino Glaxo Rijswijk (WAG-Rij) did not change the duration of SWD; however, stimulation resulted in a significant increase in the number of SWD from 18.52 ± 11.46 bursts/30 min to 30.17 ± 18.43 bursts/30 min. Whereas in control injection, the duration and the number of SWDs were similar at pre- and poststimulus. Both the background and poststimulus average firing rates of the SWD in WAG-Rij were significantly higher than the firing recorded in GAERS. Conclusion: These findings suggest that VB astrocytes play a role in modulating the SWD generation in both rat models with distinct mechanisms and can present an essential target for the possible therapeutic approach for AE.

Comparison of astrocytes and gap junction proteins in the white matter of genetic absence epileptic and control rats: an experimental study.

OBJECTIVES: The role of white matter astrocytes in absence epilepsy is unknown. The present study aims to quantify astrocytic markers glial fibrillary acidic protein (GFAP), gap junction's proteins connexin 30 (Cx30) and connexin 43 (Cx43) in the corpus callosum (CC) of genetic absence epileptic rats from Strasbourg (GAERS), Wistar albino glaxo rats from Rijswijk (WAG/Rij)and compare the results with control animals. METHODS: -The density of GFAP, Cx30 and Cx43 positive astrocytes in per unite area were quantified in the CC of GAERS, WAG/Rij and control animals using immunohistochemistry and real-time quantitative polymerase chain reaction (RT-qPCR). The quantifications were made from three regions of CC; below the primary somatosensory (S1BF), below the motor (M1) and below the retrosplenial (RSG) cortices. RESULTS: oThe number GFAP, Cx30 and Cx43 immunopositive astrocytes showed heterogeneous distribution within the CC. The GFAP immunopositive astrocytes was significantly high in the S1BF region of the three strains. The immunopositive GFAP and Cx43 showed significant decrease in the S1BF and M1 regions in GAERS and WAG/Rij compared to control animals, however, an increase in the immunopositive Cx30 was observed in the same regions in both GAERS and WAG/Rij compared to control Wistar animals but the increase was significant for GAERS but not for WAG/Rij. The RT-qPCR analysis was corroborated by GFAP immunohistochemistry results. CONCLUSION: The different expression pattern of the two Cx's in the CC of the epileptic strains compared to control animals may indicate a compensatory response or maybe the cause of generalization of absence seizures.