Exploring atypical locations of mammalian neural stem cells: the human filum terminale.

Neurogenesis is a multifactorial event determined by local environmental cues, inherent cellular program as well as cellular milieu and may not necessarily be restricted to the SVZ and SGZ. NSCs have been isolated from or neurogenesis has been demonstrated in traditionally non neurogenic regions. This more permissive view of neurogenesis, however, is not widely accepted due to concerns regarding the methodologies used. Furthermore, it is compounded by the fact that the basal levels of increased neurogenesis in such regions has not been completely confirmed and thus precludes a paradigm shift. Were this non limited view of neurogenesis to be generally accepted after thorough investigation, it would open new avenues for regenerative medicine and stem cell therapy.

Predictors of seizure outcome after temporal lobectomy for intractable epilepsy.

OBJECTIVES: To assess predictors of outcome of temporal lobectomy for intractable epilepsy. MATERIAL AND METHODS: In 63 adult patients operated with anterior temporal lobectomy during 198892, we used logistic regression analysis to assess predictors of being seizure-free (Engel's class I) 2 years after surgery. As potential predictors, we included the following variables: gender, age at operation, age at onset of seizures, epilepsy duration, etiology, generalized vs not generalized seizures, seizure frequency, intelligence quotient, ictal electroencephalography, magnetic resonance imaging (MRI), single-photon emission computed tomography (SPECT), side of resection, and extent of the resection. RESULTS: About 44% of the surgery patients were seizure-free (Engel's class I) 2 years after surgery. In multivariate analysis (n = 55), MRI pathology defined as atrophy in the temporal lobe, angioma, tumor or mesial temporal sclerosis (odds ratio, OR 7.4, 95%CI: 1.7-32.9) and extent of the hippocampal resection (increase of 1 cm) (OR 2.2, 95%CI: 1.1-4.6) predicted being seizure-free. CONCLUSION: Focal pathology in preoperative MRI and the extent of the hippocampal resection were the only significant predictors of being seizure-free after 2 years.

[Resection of malignant tumors involving the anterior cranial fossa]. / Reseksjon av maligne svulster med relasjon til fremre skallgrop.

BACKGROUND: The craniofacial approach has greatly facilitated resections of tumours involving the base of the anterior cranial fossa when compared to either the transcranial or transfacial approach alone. MATERIAL AND METHODS: This approach was used in 11 patients with malignant tumours localized to the ethmoid sinus, orbit and bone or soft tissue of the base of the anterior part of the skull. By combining a low frontal or frontolateral craniotomy with resection of the facial skull, en bloc resections were accomplished. A frontogaleal periostal flap or a muscle flap from the temporal muscle was used to replace resected bone and to seal the skull base. RESULTS: There were no peri- or postoperative deaths. One patient died due to local recurrence, one patient is alive with residual tumour six years after surgery, and one is reoperated due to local recurrence. In addition one patient developed recurrence of a previously treated tumour of the maxillary sinus. Two patients developed meningitis and one pneumocephalus postoperatively. One patient has partial loss of vision and two patients underwent dacryocystorhinostomy due to epiphora. INTERPRETATION: The planning and execution of this type of surgery requires close interaction in an interdisciplinary team, in particular between neurosurgeon and head and neck surgeon.

CTA in patients with acute subarachnoid haemorrhage. A comparative study with selective, digital angiography and blinded, independent review.

PURPOSE: Minimal- or non-invasive methods replacing intra-arterial digital subtraction angiography (IA-DSA) would be of great importance in patients suffering from acute subarachnoid haemorrhage (SAH). The aims of this study were to compare CTA with IA-DSA in patients with acute SAH, to compare CTA interpretations with those of blinded, independent reviewers and to evaluate improvement in CTA diagnostics after 1 year of experience with CTA. MATERIAL AND METHOD: During 2 years 162 patients with SAH underwent CTA as well as IA-DSA. Independent blinded review of 77 patients was performed for 1 year. RESULTS: Totally 144 aneurysms were demonstrated in 119 patients at IA-DSA, while 43 patients had normal intracranial arteries. Initially 131 aneurysms were detected at CTA while 2 normal, tortuous arteries were misinterpreted as aneurysms, giving a sensitivity of 91% and a specificity of 95%. At independent blinded review the observer agreement was 87% and the kappa value 0.68. CONCLUSION: CTA in SAH is of great value in demonstrating vascular anatomy and the exact size of an aneurysm. However, IA-DSA is still needed for diagnostic evaluation in aneurysms smaller than 5 mm in diameter, especially in those located near bony structures.

Depolarization induces insulin-like growth factor binding protein-2 expression in vivo via NMDA receptor stimulation.

The effect of depolarization and N-methyl-D-aspartate (NMDA) receptor blockade on insulin-like growth factor-I (IGF-I), IGF binding protein-2 (IGFBP-2) and IGFBP-4 expression was analysed in vivo. Depolarization was induced in adult rat brains by applying 3 M KCl to the exposed cortex for 10 min. A subgroup of animals also received daily injections of MK-801. Four days after KCl exposure, the brains were analysed by in situ hybridization, immunohistochemistry and TUNEL. A significant upregulation of IGFBP-2 mRNA and protein was detected in astrocytes after KCl exposure This upregulation was reduced by MK-801 treatment. No alterations in IGF-I or IGFBP-4 mRNA levels were noted. We did not detect TUNEL positive cells, morphological signs of necrosis or apoptosis, or neuronal loss in the depolarized zone. Taken together, these findings indicate that upregulation of IGFBP-2 by depolarization is mediated by NMDA receptors, and, as no neuronal damage was detected, astrocytic NMDA receptors may be responsible for this upregulation.

A tribute to Helge Nornes.

This supplement of the Acta Neurochirurgica is dedicated to professor Helge Nornes on the occasion of his retirement. Helge Nornes started his neurosurgical training in Oslo in 1965. In 1980 he was offered the neurosurgical chair of Bern, Switzerland, where he stayed until 1983 when his old university called him back to the chair at the National Hospital in Oslo, a position he filled until he retired last year. The present paper briefly reviews examples of his contributions to neurosurgery and to the understanding of intracranial pathophysiology, including the transcranial doppler, the miniature transducer for intracranial pressure monitoring, his observations on intracranial pressure and internal carotid blood flow during subarachnoid haemorrhage, intracranial arterial blood flow in patients undergoing aneurysm surgery, his studies of the pathophysiology of arteriovenous malformations, the introduction of intraoperative Doppler recordings during surgery for aneurysms and arteriovenous malformations, and his methods for evaluating collateral circulation prior to internal carotid artery occlusion.

Surgical treatment of anterior circulation aneurysms.

The purpose of this paper is to present the results, assessed by an independent observer, of surgical treatment of 428 consecutive patients harbouring aneurysms of the anterior circulation, together with a review of relevant anatomy and operative strategy. At follow-up (mean 5.6 years) 89.3% lived at home and were independent, 5.1% lived at home but needed some kind of assistance, 2.0% lived in institution, whereas information was unavailable in 3.6% of living patients. Two hundred and fifty-three patients (64.5%) had unchanged employment status, 0.3% worked in sheltered environment, whereas 30.9% went out of work due to their subarachnoid hemorrhage (SAH). Information about employment status was unavailable in 4.3%. For aneurysms of the internal carotid, anterior communicating and middle cerebral artery, respectively, mortality was 3.2, 3.9 and 5.6%, whereas 92.0, 88.1 and 89.0% of surviving patients lived at home and were independent and 67.0, 63.6 and 63.0% had unchanged employment status. Three-months mortality of all causes was 4.2%. In the postoperative period 53 (12.4%) patients developed clinical signs of vasospasms, 6 (1.4%) had cardiac infarction, 4 (0.9%) lung oedema, 4 (0.9%) deep vein thrombosis, and 7 patients (1.6%) infection. During the follow-up period shunt-dependent hydrocephalus developed in 4.2% and 0.2% had a subsequent SAH from the same aneurysm. Forty-three patients were on anticonvulsive therapy.

Depletion of intracellular Ca2+ stores or lowering extracellular calcium alters intracellular Ca2+ changes during cerebral energy deprivation.

Cytoplasmatic calcium concentrations are elevated two to three fold during cerebral ischemia. In order to determine the role of calcium-release from intracellular stores vs. calcium entry from the extracellular space, intracellular stores were depleted by the use of thapsigargin and calcium was removed from the incubation fluid prior to energy deprivation (ED). CA 1 pyramidal neurons in hippocampal rat slices were filled with a 1:2 mixture of Fluo-3 and Fura Red by intracellular injection. The neurons were visualized in a Confocal Laser Scanning Microscope (CLSM) and the fluorescence ratio from the probe mixture was used to quantify the calcium concentration. Intracellular calcium concentration was monitored before and during ED. The intracellular calcium concentration was 55 nM prior to ED and increased to 25 microM during ED. The resting levels were the same in the experimental groups, but the increase during ED was significantly lower in the intervention groups. The increase in the calcium free group was to 1 microM and in the thapsigargin group to 5 microM. In the last experimental group, thapsigargin treatment and removal of extracellular calcium, the intracellular calcium increased to 630 nM. These results demonstrate that the increased intracellular calcium seen during ED originates from several sources. Calcium-release from intracellular stores may be of major importance in calcium-related neuronal injury during cerebral ischemia.

Effect of isoflurane on release and uptake of gamma-aminobutyric acid from rat cortical synaptosomes.

We have studied the effect of isoflurane on potassium-evoked release and high-affinity uptake of gamma-aminobutyric acid (GABA) in rat cortical synaptosomes. Isoflurane 1.5% and 3% increased calcium-dependent release by 38% and 36% of control values, respectively (P < 0.05). Calcium-independent release was reduced correspondingly by 24% and 26% (P < 0.05). High-affinity uptake of GABA was not affected by isoflurane. The findings of increased synaptic GABA release combined with unaltered uptake suggest that isoflurane increases GABA in the synaptic cleft and thus may enhance inhibition.

The effect of isoflurane on brain amino acid release and tissue content induced by energy deprivation.

This article describes the effect of isoflurane on amino acid release and tissue content induced by energy deprivation in slices of rat hippocampus. Energy deprivation (95% N2 / 5% CO2 and glucose free medium) (ED) induced an increase in the release of all amino acids measured, with the exception of glutamine. The tissue content of all amino acids except gamma-aminobutyric acid (GABA) and arginine was concomitantly reduced. Isoflurane (1.5% and 3.0%) reduced glutamate release during ED by 27% and 28% (p < 0.05 as compared with release without isoflurane), respectively, whereas the tissue content was slightly increased. Similarly, GABA release was reduced by 25% and 25% (p < 0.05 as compared with release without isoflurane) accompanied by an insignificant enhancement in tissue content as compared with ED without isoflurane. Isoflurane reduced the release of taurine and most of the other amino acids. The total amount of all amino acids (both released and retained) was not significantly altered by the anesthetic. These observations demonstrate that isoflurane can modify the changes in amino acid handling induced by energy deprivation.

Confocal laser scanning microscopy used to monitor intracellular Ca2+ changes in hippocampal CA 1 neurons during energy deprivation.

An increase in intracellular calcium during cerebral ischemia has been proposed as a common final pathway underlying the events leading to neuronal death. Intracellular calcium has been measured with ion selective electrodes during energy deprivation (ED) in hippocampal slices and with fluorescent techniques in neuronal cultures. In the present study, we describe a novel method to visualize and quantify changes in intracellular calcium in brain slices using Confocal Laser Scanning Microscopy (CLSM). CA 1 pyramidal neurons in hippocampal slices were filled by intracellular injection with a 1:2 mixture of the fluorescent dyes Fluo 3 and Fura Red. The neurons were then visualized using CLSM, and the ratio of the fluorescence from each probe used to quantify intracellular calcium concentrations before and during ED. The free intracellular calcium concentration was 60 nM prior to ED and increased to 24 microM during ED. These results demonstrates that CLSM and fluorescent probes can be used in functional neuronal networks in addition to cell cultures as previously described.

Isoflurane reduces synaptic glutamate release without changing cytosolic free calcium in isolated nerve terminals.

The molecular mechanism of volatile anaesthetic action on presynaptic glutamate release is not clear. An inhibitory effect on voltage-gated calcium channels has been proposed. The present study examines the effect of isoflurane on cytosolic free calcium and synaptic glutamate release from isolated nerve terminals. Synaptosomes from rat cerebral cortex were used. Glutamate was measured with a continuous fluorometric measurement in a spectrophotometer as the fluorescence of NADPH and calcium as the fluorescence of fura-2. Isoflurane reduced the calcium-dependent glutamate release evoked by membrane depolarization with 4-aminopyridine in an inversely dose-dependent manner. The glutamate release was reduced by 56, 43 and 36% in response to isoflurane 0.5, 1.5 and 3.0%, respectively (for all: P < 0.05). Membrane depolarization evoked a rise in cytosolic free calcium of approximately 34%. Addition of isoflurane (0.5, 1.5 and 3.0%) produced no significant change in cytosolic free calcium. These results indicate that the isoflurane-induced reduction in presynaptic glutamate release is caused by other mechanisms than blocking voltage-gated calcium channels. As the release is inversely dose-dependent, two or more mechanisms could be involved.

Chloride influx during cerebral energy deprivation.

The purpose of the present study was to investigate the possible role of chloride influx and GABA release during cerebral energy deprivation (ED). The functional activity measured by evoked activity (population spike) in hippocampal slices was recorded during nine minutes of ED and 60 minutes recovery. Treatment groups were exposed to ED following administration of the GABAA antagonist penicillin G (pc G) or substitution of extracellular chloride. The release of glutamate and GABA was measured by HPLC. The efflux of 36Cl from preloaded slices was measured during ED with and without blocking the GABAA receptor. The population spike disappeared during ED, and there was a marked release of GABA and glutamate. During recovery the population spike recovered partially. Both application of pc G and substitution of extracellular chloride during ED improved population spike recovery. Uptake of radiolabeled chloride was significantly reduced by pc G. Glutamate release, but not GABA, was significantly reduced by chloride substitution. These results indicate a possible role of chloride mediated injury during ED, and suggest that chloride entry may partly occur through ligand-operated channels. Furthermore there may be an early chloride dependent release of glutamate during cerebral ischemia, whereas later release seems to be chloride independent.

The effect of volatile anaesthetics on synaptic release and uptake of glutamate.

1. Volatile anaesthetics seem to exert their effects on several parts of the neuronal conducting system. 2. The effect on synaptic excitation seems to be quantitatively the most important (Berg-Johnsen and Langmoen, Acta Physiol. Scand. 128, 1986, 613-618) as 1 minimum alveolar concentration (MAC) of isoflurane reduces the activity in thin unmyelinated afferent fibres by 18%, excitatory synapses by 27% and postsynaptic neurones by 24%. 3. The reduction in excitatory synaptic transmission is caused by a decreased amount of transmitter glutamate in the synaptic cleft caused by a reduced release and increased uptake of glutamate in the presynaptic terminals (Larsen et al., Brain Res. 663, 1994, 335-337; Larsen et al., Br. J. Anaesth. 78, 1997, 55-59).

L-alpha-aminoadipate reduces glutamate release from brain tissue exposed to combined oxygen and glucose deprivation.

The effect of the glutamate analogue L-alpha-aminoadipate (alpha AA) on the release of glutamate and gamma-aminobutyric acid (GABA) from rat hippocampal slices was investigated in vitro. Oxygen/glucose deprivation caused a large release of glutamate and GABA. alpha AA added during energy deprivation reduced the glutamate release in a dose-dependent manner (56% reduction at 5 mM), whereas GABA release was unchanged. We speculate that ischemic glutamate release from the brain is mediated by a low affinity transport mechanism that is blocked by alpha AA.

Efflux of gamma-aminobutyric acid caused by changes in ion concentrations and cell swelling simulating the effect of cerebral ischaemia.

The relationships among ischaemic GABA efflux from brain tissue and extracellular and intracellular concentrations of sodium, chloride and potassium ions were investigated by means of 1) transverse hippocampal slices from rat and 2) functional expression of a high affinity GABA transporter in Xenopus oocytes. Brain slices were incubated for 20 min in medium where extracellular sodium and chloride were substituted with impermeant ions. Isethionate (Iseth) substitution for chloride generated a 7-fold increase in GABA efflux. Choline (Chol) but not N-methyl-D-glucamine (NMDG) substitution for sodium likewise increased GABA efflux. Reducing the osmolarity of the medium by decreasing both sodium and chloride concentrations (Hyp) increased GABA efflux 3-fold. This release was blocked by mannitol (Man). Blocking sodium channels with 1 microM of tetrodotoxin (TTX) also increased the release 3-fold. Energy deprivation (ED) increased the GABA release 50-fold. ED/Iseth left the release unchanged, ED/Chol increased the GABA efflux by 23%, whereas ED/NMDG reduced the release by 41%. Adding mannitol did not block the ED-evoked release, whereas TTX reduced it by 52%. Release of preloaded [3H]-GABA from oocytes expressing the GAT-1 GABA transporter was then examined. Depolarisation by current injection or 100 mM extracellular K+ did not increase GABA release. Sodium chloride injection, however, caused membrane depolarisation and a 100-fold increased GABA efflux from the oocytes. This release was blocked when the osmolarity was increased extracellularly by adding mannitol. These results show that 1) TTX releases GABA from brain tissue but blocks release during ED, 2) the high affinity GABA carrier must be altered in order to reverse, 3) ischaemic GABA release is sodium independent, and is modulated by large cations, 4) mannitol blocks the reversal of high affinity carriers in oocytes, but the release from brain slices during ED is unaffected. Taken together, the results suggest that ischaemic release of GABA from brain tissue does not occur by means of reversed high affinity carriers alone, but rather that it is controlled by more complex mechanisms.

Isoflurane increases the uptake of glutamate in synaptosomes from rat cerebral cortex.

We have studied the effect of increasing concentrations of isoflurane on high- and low-affinity uptake of L-glutamate using synaptosomes from rat cerebral cortex. In the high-affinity uptake range, 0.5% isoflurane had no effect on uptake velocity, while 1.5% and 3.0% isoflurane caused an increase in mean Vmax to 131 (SEM 54) and 210 (103)% of control, respectively. There was no significant change in the K(m) value. Vmax and K(m) values for low-affinity uptake of L-glutamate were unchanged by 1.5% isoflurane. These results provide evidence for an isoflurane-induced increase in high-affinity uptake of glutamate into presynaptic terminals. This effect may contribute to a reduction of transmitter in the synaptic cleft and thereby decreased excitatory synaptic transmission.

Cytotoxic effect of Ca++ released from intracellular stores during cerebral energy deprivation.

The increase in cytoplasmatic calcium concentration during cerebral ischemia has been proposed as a key event leading to neuronal death. In order to investigate a possible role of calcium-release from intracellular stores in ischemic neuronal injury, intracellular calcium pools were depleted prior to ischemia by the use of thapsigargin. Evoked activity (population spike) in rat hippocampal slices was monitored during a 30 min control period, 9 min of energy deprivation and 60 min of recovery. The population spike recovered to 27% (17-33) (median and 95% confidence interval) following energy deprivation in normal calcium, to 56% (50-58) in calcium-free incubation fluid and to 83% (75-88) in slices pretreated with 1 microM thapsigargin. Combining calcium removal and thapsigargin pretreatment did not improve recovery further. Both removal of extracellular calcium and emptying intracellular calcium stores prior to energy deprivation thus improved functional recovery following energy deprivation, however the latter was more effective. These results suggest that calcium release from intracellular stores may be of major importance in calcium-related neuronal injury during cerebral ischemia.