Biomedical subjects

E T MacKenzie

Publications and source records attributed to E T MacKenzie.

At least 37 records · Page 2Linked to original sources

PubMed1999

Transforming growth factor-beta1 as a regulator of the serpins/t-PA axis in cerebral ischemia.

The tissue type plasminogen activator (t-PA) is a serine protease that is involved in neuronal plasticity and cell death induced by excitotoxins and ischemia in the brain. t-PA activity in the central nervous system is regulated through the activation of serine protease inhibitors (serpins) such as the plasminogen activator inhibitor (PAI-1), the protease nexin-1 (PN-1), and neuroserpin (NSP). Recently we demonstrated in vitro that PAI-1 produced by astrocytes mediates the neuroprotective effect of the transforming growth factor-beta1 (TGF-beta1) in NMDA-induced neuronal cell death. To investigate whether serpins may be involved in neuronal cell death after cerebral ischemia, we determined, by using semiquantitative RT-PCR and in situ hybridization, that focal cerebral ischemia in mice induced a dramatic overexpression of PAI-1 without any effect on PN-1, NSP, or t-PA. Then we showed that although the expression of PAI-1 is restricted to astrocytes, PN-1, NSP, and t-PA are expressed in both neurons and astrocytes. Moreover, by using semiquantitative RT-PCR and Western blotting, we observed that only the expression of PAI-1 was modulated by TGF-beta1 treatment via a TGF-beta-inducible element contained in the PAI-1 promoter (CAGA box). Finally, we compared the specificity of TGF-beta1 action with other members of the TGF-beta family by using luciferase reporter genes. These data show that TGF-beta and activin were able to induce the overexpression of PAI-1 in astrocytes, but that bone morphogenetic proteins, glial cell line-derived neutrophic factor, and neurturin did not. These results provide new insights into the regulation of the serpins/t-PA axis and the mechanism by which TGF-beta may be neuroprotective.

Amyloid beta-Protein Precursor↗

PubMed1999

A potential role for erythropoietin in focal permanent cerebral ischemia in mice.

The present study describes, for the first time, a temporal and spatial cellular expression of erythropoietin (Epo) and Epo receptor (Epo-R) with the evolution of a cerebral infarct after focal permanent ischemia in mice. In addition to a basal expression of Epo in neurons and astrocytes, a postischemic Epo expression has been localized specifically to endothelial cells (1 day), microglia/macrophage-like cells (3 days), and reactive astrocytes (7 days after occlusion). Under these conditions, the Epo-R expression always precedes that of Epo for each cell type. These results support the hypothesis that there is a continuous formation of Epo, with its corresponding receptor, during the active evolution of a focal cerebral infarct and that the Epo/Epo-R system might be implicated in the processes of neuroprotection and restructuring (such as angiogenesis and gliosis) after ischemia. To support this hypothesis, a significant reduction in infarct volume (47%; P < 0.0002) was found in mice treated with recombinant Epo 24 hours before induction of cerebral ischemia. Based on the above, we propose that the Epo/Epo-R system is an endogenous mechanism that protects the brain against damages consequent to a reduction in blood flow, a mechanism that can be amplified by the intracerebroventricular application of exogenous recombinant Epo.

Animals↗

PubMed1999

A transforming growth factor-beta antagonist unmasks the neuroprotective role of this endogenous cytokine in excitotoxic and ischemic brain injury.

Various studies describe increased concentrations of transforming growth factor-beta (TGF-beta) in brain tissue after acute brain injury. However, the role of endogenously produced TGF-beta after brain damage to the CNS remains to be clearly established. Here, the authors examine the influence of TGF-beta produced after an episode of cerebral ischemia by injecting a soluble TGF-beta type II receptor fused with the Fc region of a human immunoglobulin (TbetaRIIs-Fc). First, this molecular construct was characterized as a selective antagonist of TGF-beta. Then, the authors tested its ability to reverse the effect of TGF-beta1 on excitotoxic cell death in murine cortical cell cultures. The addition of 1 microg/mL of TbetaRIIs-Fc to the exposure medium antagonized the neuroprotective activity of TGF-beta1 in N-methyl-D-aspartate (NMDA)-induced excitotoxic cell death. These results are consistent with the hypothesis that TGF-beta1 exerts a negative modulatory action on NMDA receptor-mediated excitotoxicity. To determine the role of TGF-beta1 produced in response to brain damage, the authors used a model of an excitotoxic lesion induced by the intrastriatal injection of 75 nmol of NMDA in the presence of 1.5 microg of TbetaRIIs-Fc. The intrastriatal injection of NMDA was demonstrated to induce an early upregulation of the expression of TGF-beta1 mRNA. Furthermore, when added to the excitotoxin, TbetaRIIs-Fc increased (by 2.2-fold, P < 0.05) the lesion size. These observations were strengthened by the fact that an intracortical injection of TbetaRIIs-Fc in rats subjected to a 30-minute reversible cerebral focal ischemia aggravated the volume of infarction. In the group injected with the TGF-beta1 antagonist, a 3.5-fold increase was measured in the infarction size (43.3 +/- 9.5 versus 152.8 +/- 46.3 mm3; P < 0.05). In conclusion, by antagonizing the influence of TGF-beta in brain tissue subjected to excitotoxic or ischemic lesion, the authors markedly exacerbated the resulting extent of necrosis. These results suggest that, in response to such insults, brain tissue responds by the synthesis of a neuroprotective cytokine, TGF-beta1, which is involved in the limitation of the extent of the injury. The pharmacologic potentiation of this endogenous defensive mechanism might represent an alternative and novel strategy for the therapy of hypoxic-ischemic cerebral injury.

Animals↗

PubMed1999

Differential time-course decreases in nonselective, mu-, delta-, and kappa-opioid receptors after focal cerebral ischemia in mice.

BACKGROUND AND PURPOSE: Neuroprotection studies have demonstrated the involvement of opioids in ischemia, and we have previously demonstrated alterations in Bmax of opioidergic receptors after 2 post-MCAO time points in mice. METHODS: In the present study, we have investigated in a detailed manner the postischemic time course of variations in [3H]diprenorphine (nonselective), [3H]DAMGO (mu), [3H]DADLE (delta), and [3H]U69593 (kappa) relative binding densities after focal cerebral ischemia (0 to 48 hours) in mice. RESULTS: In frontoparietal cortices, our results demonstrate decreases in (1) delta receptor densities at 1 to 3 hours after MCAO, (2) mu and nonselective binding sites at 6 to 12 hours after MCAO, and (3) kappa receptor densities between 6 and 24 hours after MCAO. In the rostral part of the infarct border zone, a decrease in delta-receptors was found concomitant with the extension of the infarct core; conversely, the decrease in delta-receptors appeared before (6 to 12 hours) macroscopic histological damage, which occurred between 12 hours and 24 hours after MCAO in the caudal part of this area. In this frontier, mu- and especially kappa-binding sites were decreased later (12 to 48 hours after MCAO). CONCLUSIONS: These differential alterations in opioidergic receptors could be due to the selective sublocalization of receptors, postsynaptically on cortical interneurons for mu- and delta-receptors versus presynaptically on cortical afferent pathways for the kappa subtype. Further, our results suggest that delta- and mu-opioidergic receptors could be markers of infarct extension and neuronal death; the study of [3H]diprenorphine and selective binding sites argues in favor of the use of receptor-specific ligands. Finally, the relative preservation of kappa-receptors might be correlated with the neuroprotective role of kappa-agonists, as previously reported.

Animals↗

PubMed1998

Autoradiographic mapping of cerebral blood flow responses to cholinergic stimulation of the rat substantia innominata: modulatory effect of galanin.

In order to analyze the precise cerebrovascular effects of a specific cholinergic stimulation of the rat substantia innominata and their modulation by galanin, cerebral blood flow was measured by the [14C]-iodoantipyrine autoradiographic method in anesthetized (urethane and alpha-chloralose), artificially ventilated male Sprague-Dawley rats that received a microinjection into the substantia innominata of saline (n = 7), or 63 pmol of galanin (n = 8), or 50 nmoles of carbachol (n = 6) or a coinjection of carbachol and galanin (n = 8). Significant carbachol-induced cerebral blood flow increases were noted in ipsilateral cortices (+36%, p < 0.01 in the cingulate to +82%, p < 0.01 in the parietal somatosensory cortices), but also in ipsilateral hippocampus and ipsilateral thalamus. These cerebral blood flow increases were abolished by the coinjection of carbachol and galanin, while infusions of galanin alone failed to affect cerebral blood flow. Cholinergic stimulation of the substantia innominata represents thus a good model for the analysis of the detailed pharmacological properties of the cholinergic vasodilatatory basalocortical system. The existence of an inhibitory galaninergic modulation of this system could be of particular interest, in terms of cerebrovascular reactivity, in various neurodegenerative states.

Animals↗

PubMed1998

Maximal densities of mu, delta, and kappa receptors are differentially altered by focal cerebral ischaemia in the mouse.

Though opioids are known to have neuroprotective properties, little information is available on the functional state of opioidergic receptors following focal cerebral ischaemia. The present study investigated the evolution of the Bmax and Kd for [3H]DAMGO, [3H]DADLE, and [3H]U69,593, respectively, for the mu, delta, and kappa opioidergic receptors after permanent focal cerebral ischaemia in mice. While the various Kd were unchanged, mu and delta Bmax values were precociously decreased in frontoparietal cortices, earlier than kappa receptors, reflecting infarct extension with time. The Bmax values for mu and delta receptors were also altered in non-infarcted tissues, such as tissues at risk (e.g., temporal auditory cortex) and exofocal (e.g., contralateral and non-infarcted) cortices. These results suggest that, in non-infarcted areas, the observed changes reflect functional modifications to focal ischaemia.

Animals↗

PubMed1998

Selective neuronal vulnerability and specific glial reactions in hippocampal and neocortical organotypic cultures submitted to ischemia.

Neurons from cerebral neocortex and hippocampus exhibit a striking difference in vulnerability to transient global ischemia. In order to study the contribution of neuronal connections and neuron-glia interactions to this variation in neuronal vulnerability, we used hippocampal and neocortical cultures submitted to various periods of histotoxic ischemia. Organotypic cultures were exposed at 37 degrees C for 0, 7, 30 and 60 min to a glucose-free NaCN-containing medium. Histological analysis using thionin staining and MAP2 immunostaining showed differences in the temporal profile of neuronal damage in hippocampal and neocortical structures, i.e., in decreasing order, CA1 (7 min) > CA3 and neocortical layers II, III, V, VI (30 min) > DG and neocortical layer IV (60 min). In parallel to the neurodegeneration study, the time course and the regional pattern of microglial and astroglial changes were also examined using GS-B4 isolectin and GFAP as immunohistochemical markers, respectively. The GS-B4 isolectin staining revealed an early (at 7 min for the hippocampus) and a specific microglial activation located in areas undergoing neuronal damage. For both organotypic cultures, astrogliosis occurred later (after 30 min of stress) with no specific regional distribution. Both hippocampal and neocortical cultures submitted to histotoxic ischemia allowed the replication of many of the cellular events observed in response to global ischemia in vivo. These findings support the hypothesis that neuron-neuron connections as well as interactions between neurons and glial cells are essential to reproduce in vitro the selective neuronal vulnerability described in vivo.

Adaptation, Physiological↗

PubMed1998

Evidence of type I and type II transforming growth factor-beta receptors in central nervous tissues: changes induced by focal cerebral ischemia.

The peptides of the transforming growth factor-beta (TGF-beta) family transduce their signal through ligand-induced heteromeric complexes that consist of type I and type II serine/threonine kinases. Both TGF-beta receptors are abundant in many peripheral tissues, but clear evidence of their expression in cortical astrocytes and neurons has not been published so far. In this study, we investigated the expression of type I and type II TGF-beta receptors and their potential ligands (TGF-beta1, TGF-beta2, and TGF-beta3) in the CNS by using RT-PCR and immunohistochemistry. Moreover, to further the study of those cell types that exhibit TGF-beta isoforms and related receptors, we examined through the use of RT-PCR whether cortical neurons and astrocytes in culture express the mRNAs for TGF-betas and their receptors. We show that the three TGF-beta isoform mRNAs are present in the CNS. However, although astrocytes in culture display all three isoforms, neurons in culture express only TGF-beta2. We have demonstrated that both type I and type II TGF-beta receptor mRNAs and proteins are present in the CNS and in cultures of cortical neurons and astrocytes. Thus, TGF-betas may act as autocrine and paracrine signals in the CNS between both neurons and astrocytes via the same receptor systems as those found in peripheral tissues. TGF-beta1 has been shown to be induced following hypoxic-ischemic brain injury and may play a critical role in the pathophysiology of degenerative processes in the CNS. In the present investigation, we confirmed that the expression of TGF-beta1 was increased markedly up until 24 h and thereafter was stable over the first 3 days following permanent occlusion of the middle cerebral artery in mice. However, whereas the expression of the type I TGF-beta receptor was not altered by the ischemic insult, the pattern of the type II TGF-beta receptors was modified dramatically in the ischemic area 3 days after the occlusion. These data show that, even if ligands are present, they may not be able to transduce their signal. Finally, the present study clearly demonstrates that a knowledge of the expression of ligand-specific receptors following brain injury is a fundamental step in clarifying the involvement of cytokines in neurodegenerative diseases.

Activin Receptors, Type I↗

PubMed1998

Evaluation of the ratio of cerebral blood flow to cerebral blood volume as an index of local cerebral perfusion pressure.

Local cerebral perfusion pressure (CPP), a crucial parameter that should allow a better assessment of the haemodynamic compromise in cerebrovascular diseases, is not currently measurable by non-invasive means. Experimental and clinical studies have suggested that the regional ratio of cerebral blood flow to cerebral blood volume (CBF:CBV), as measured by PET, represents an index of local CPP in focal ischaemia. The present study was designed to evaluate further the reliability of the CBF:CBV ratio during manipulations of CPP by deliberately varying mean arterial pressure (MAP) in the anaesthetized baboon. Cortical CBF, CBV, cerebral metabolic rate for oxygen (CMRO2) and oxygen extraction fraction were measured by PET using the (15)O steady-state technique in 10 anaesthetized baboons. Five baboons (Group A) underwent four PET examinations at different levels of MAP: base line (101 +/- 6 mmHg) followed by moderate hypotension (58 +/- 3 mmHg) and, in a separate experiment, minor hypotension (72 +/- 3 mmHg) followed by profound hypotension (34 +/- 5 mmHg). Trimetaphan was used to lower MAP to minor and moderate levels while profound hypotension was achieved by the combined effects of trimetaphan and lower-body negative pressure. Five other baboons (Group B) were subjected to hypertension (121 +/- 2 mmHg) induced by metaraminol and were compared with their base line state (81 +/- 10 mmHg). While CBF displayed significant changes with varying MAP, i.e. decrease and increase with hypotension and hypertension, respectively (-11% from base line to moderate hypotension compared with -20%, from minor to profound hypotension and +31% from base line to hypertension), CBV was more variable and did not significantly change, except with profound hypotension when the increase was significant (+13%). The CBF:CBV ratio decreased significantly at all stages of hypotension (-21 and -31%) and was significantly increased during hypertension (+30%). Importantly, the CBF:CBV ratio demonstrated a significant correlation with MAP (rho = 0.78, Spearman's rank correlation coefficient, P < 0.01). No major changes in CMRO2 were noted during either hypotension or hypertension. Our results demonstrate that, under physiological conditions, cortical CBF:CBV is significantly correlated with CPP, itself a function of MAP. In the investigated range of MAP, the relationships between CBF:CBV and MAP appear to be linear. These findings further argue for the reliability of CBF:CBV as an index of CPP in situations where increases or decreases of MAP without superimposed changes in cerebrovascular tone are encountered, and they confirm the potential usefulness of this regional ratio for clinical investigations and management in cerebrovascular diseases.

Animals↗

PubMed1997

Changes in ouabain affinity of Na+, K+-ATPase during focal cerebral ischaemia in the mouse.

We investigated the effect of focal cerebral ischaemia on the activity and the affinity of the ouabain sites of Na+,K+-ATPase in the mouse. The Na+,K+-ATPase activity was decreased by 38% as early as 30 min following ischaemia. In the sham group, the dose-response curves for ouabain disclosed three inhibitory states which contribute, respectively, 24.9 +/- 6.7%, 39.1 +/- 7.5% and 36.0% of the total activity (low affinity, LA; high affinity, HA and very high affinity, VHA, respectively). Their computed IC50 values are, respectively: 1.3 X 10(-3) M, 4.5 X 10(-6) M and 2.9 X 10(-9) M. Surprisingly, in ischaemic cortices, only two sites for ouabain were detected. The first site exhibits a LA (IC50 = 2.0 X 10[-4] M) but its relative contribution to the total activity (46.1 +/- 5.2%) is twice that noted for the LA site in non-ischaemic tissues. The second site presents an affinity intermediate between those of HA and VHA sites of the sham group (IC50 = 1.7 X 10[-7] M) and contributes 53.9% to the total activity. Loss in the specific activity of the second site explains that of the total activity. The most likely explanation in the presence of only two ouabain sites of Na+,K+-ATPase following ischaemia may be a change in ouabain affinity of alpha2 and/or alpha3 isoforms, as the presence of all three alpha isoforms has been observed by Western blotting. These results suggest that ischaemia induces intrinsic modifications in Na+,K+-ATPase which result from perturbations in membrane integrity and/or association of the alpha isoforms of this enzyme.

Animals↗

PubMed1997

Progressive impairment of brain oxidative metabolism reversed by reperfusion following middle cerebral artery occlusion in anaesthetized baboons.

A better understanding of the temporospatial evolution of ischaemic brain tissue towards necrosis would be of crucial value to establish and validate therapeutic strategies for stroke in man. By means of sequential positron emission tomographic (PET) studies performed through the acute to the chronic stages of infarction, we addressed the question of the effect of 6 h temporary occlusion of the middle cerebral artery (MCAO) on the evolution of the volume of severely hypometabolic tissue in anaesthetized baboons and compared it to that reported previously in permanently occluded baboons. Thirteen anaesthetized baboons underwent serial PET (15O steady-state technique) examinations before and 1, 4, 7, 24-48 h and 15-62 days following transorbital MCAO. Reperfusion, at 6 h post-occlusion, was assessed by Doppler sonography and cerebral blood flow (CBF) values after clip removal. In each baboon, the infarct volume was calculated by standard histological procedures 20-91 days after MCAO. The severely hypometabolic tissue volume, as defined by a threshold of oxidative metabolism, showed a progressive increase for up to 24-48 h in a not dissimilar manner to that found in baboons with permanent occlusion. However, these hypometabolic volumes regressed in the chronic stage (p < 0.05). Permanent and temporary occluded baboons, when taken together, showed a highly significant relationship between histological and chronic hypometabolic volumes (r = 0.84; p < 0.001). Moreover, the final hypometabolic volume where cerebral metabolic rate of oxygen (CMRO2) was below 40% of contralateral metabolism corresponded well to that of histological infarction volume. We conclude that, in anaesthetized baboons, restoration of blood flow will reverse (even if not immediately) the progressive derangement of metabolism after MCAO and markedly limit the final volume of consolidated infarction.)

Analysis of Variance↗

PubMed1997

Early reperfusion in the anesthetized baboon reduces brain damage following middle cerebral artery occlusion: a quantitative analysis of infarction volume.

BACKGROUND AND PURPOSE: Because in humans the clinical benefits of reperfusion remain controversial, it is important to determine whether reperfusion per se reduces infarct volume. In the nonhuman primate, mostly semiquantitative assessments of infarction have been performed. When ischemic volumes have been calculated, it has been for the acute or subacute stages of experimental stroke and may thus not adequately reflect the total volume of consolidated infarction. METHODS: Anesthetized baboons were subjected to 6 hours of either reversible or permanent middle cerebral artery occlusion (MCAO). Approximately 4 weeks later, the brains were processed for neuropathological examination to allow assessment of the final infarct volume determined by the difference of healthy tissue between occluded and nonoccluded hemispheres. RESULTS: Reversible MCAO resulted in a small essentially subcortical infarction (mean+/-SD, 0.58+/-0.31 cm3) in 6 of 10 baboons: the infarct (pannecrosis) was restricted to the head of the caudate nucleus, internal capsule, and putamen; 4 of 10 baboons showed no evidence of macroscopic infarction. Permanent MCAO produced a larger subcortical infarct in all 7 baboons studied (2.37+/-1.32 cm3; P=.0006 by Wilcoxon-Mann-Whitney test); the lesion was more extensive and encompassed the external capsule and, in 2 baboons, the adjacent insular cortex. CONCLUSIONS: We conclude that under optimal experimental conditions, an ischemic episode of 6 hours in duration is well tolerated in the anesthetized adolescent baboon, with 4 animals showing no signs of macroscopic brain damage. Thus, early reestablishment of cerebral blood flow after a focal ischemic insult is not detrimental but indeed is beneficial in terms of the final infarct volume (both at the subcortical and cortical levels) produced by occlusion of a major cerebral artery. The data further suggest a feasible time window in which to initiate and continue therapeutic interventions.

Anesthesia↗

PubMed1996

Effects of indomethacin on cerebral blood flow and oxygen metabolism: a positron emission tomographic investigation in the anaesthetized baboon.

The aim of this study was to clarify the controversy about the effects of indomethacin on the coupling of cerebral blood flow (CBF) to cerebral oxygen metabolism (CMRO2). CBF, blood volume (CBV), oxygen extraction fraction (OEF) and CMRO2 were measured by positron emission tomography (PET) in five anaesthetized baboons before and during an i.v. administration of indomethacin (bolus 20 mg/kg followed by perfusion 10 mg/kg.h). Administration of indomethacin resulted in a marked and homogenous decrease of CBF in every region analysed (-28% to -40%) and a moderate reduction in CBV (-8% to -16%). In contrast, CMRO2 displayed a small increase in thalamus and pons (+10% and +13%, respectively). OEF increased greatly in all structures studied (+59% to +96%). These findings show that the potent cerebrovascular effects of indomethacin are not related to a decrease in CMRO2 as measured through the use of PET.

Animals↗

PubMed1996

In vivo binding, pharmacokinetics and metabolism of the selective M2 muscarinic antagonists [3H]AF-DX 116 and [3H]AF-DX 384 in the anesthetized rat.

The pharmacokinetics, in vivo binding and metabolism of two M2 muscarinic receptor antagonists, [3H]AF-DX 116 and [3H]AF-DX 384, were studied in anesthetized rats, which received either the tracer alone or following a saturating injection of atropine. Both radioligands were cleared from the circulation with distribution half-lives of 17 and 14 sec and elimination half-lives of 17 and 40 min for [3H]AF-DX 116 and [3H]AF-DX 384, respectively. A radioactive distribution, predominant in peripheral organs when compared to brain, was found at each time studied after tracer injection. Atropine-displaceable tracer uptake was evidenced at 20-40 min in brain (31%), submandibular glands (26%), spleen (37%) and notably heart (55%) for [3H]AF-DX 116 but only in heart (50%) for [3H]AF-DX 384 at 10-20 min. Regional brain sampling revealed a relatively uniform distribution of [3H]AF-DX 384 and a -45% atropine saturation effect (i.e., specific binding) in the thalamus 20 min after injection. Sequential thin-layer chromatographic studies performed on tissue extracts demonstrated the rapid appearance of labeled metabolites of both radiotracers in brain (but less so in liver) and especially in cardiac tissues, where almost 70% of total radioactivity still corresponded to authentic tracer 40 min after injection. Thus, based on their low blood-brain barrier permeability and the high presence of labeled metabolites in the central nervous system, AF-DX 116 and AF-DX 384 might be more helpful in the study of M2 muscarinic receptors present in heart rather than brain. Labeled with positron emittors, these M2 antagonists might be applicable to the pathophysiological study of disease states, such as cardiomyopathies.

Anesthesia↗

PubMed1996

Unilateral eyeball enucleation differentially alters AMPA-, NMDA- and kainate glutamate receptor binding in the newborn rat brain.

The aim of the present work was to evaluate the neurochemical effects of early unilateral visual deprivation as a model of impaired visual maturation. For this purpose, binding to the different ionotropic glutamate receptor subtypes was quantified in vision-related and vision-unrelated brain structures of control and unilaterally deprived newborn rats. At post-natal (PN) day 10, male Sprague-Dawley rats underwent either unilateral eyeball enucleation (enucleation group, n = 12) or sham operation (control group, n = 12). In each group, brains were obtained either at post-natal day 20 (n = 6) or post-natal day 30 (n = 6) and processed for quantitative in vitro autoradiography selective for NMDA, kainate, and AMPA glutamate-binding sites, as well as for the presynaptic adenosine A1 receptor as a control of the deafferentation efficacy. In control animals, quantitative autoradiography revealed an increase in NMDA (e.g. +45% in superior colliculus) and kainate receptor binding (e.g. +55% in visual cortex, layer IV) from post-natal day 20 to post-natal day 30, associated with stable levels of AMPA receptor binding, in the vision-related structures. In the deafferented visual structures, monocular enucleation induced a marked decrease in A1 site density (e.g. -38 to -52%, in the superficial layer of superior colliculi, at PN day 20 and PN day 30, respectively) in parallel with a mild increase in both NMDA (e.g. +8 to 9%, in superior colliculi and visual cortex, layer IV at PN day 30, respectively) and AMPA (e.g. +16%, in layer IV of the visual cortex at PN day 30). Superimposed on marked bilateral decreases at PN day 30 in the enucleated rats, kainate receptor binding also revealed a slight but significant decrease (-5%) in the deafferented superior colliculus as compared to the non-deafferented side. The present findings (different time-courses of, and differential effects of deafferentation on, the NMDA, kainate and AMPA glutamate receptor subtypes throughout the visual brain structures) further support the involvement of these receptors in distinctive roles during maturation of the visual system.

Animals↗

PubMed1996

Relationships between high oxygen extraction fraction in the acute stage and final infarction in reversible middle cerebral artery occlusion: an investigation in anesthetized baboons with positron emission tomography.

Studies in humans suggest that regions that show maximal increases in brain oxygen extraction fraction (OEF) in the hours following an ischemic episode are those most vulnerable for infarction and are often, although not always, associated with the final site of infarction. To clarify this issue, we followed the hemodynamic and metabolic characteristics of regions with an initially maximally increased OEF and compared them with the ultimately infarcted region in an experimental stroke model. Positron emission tomography (PET) was used to obtain functional images of the brain prior to and following reversible unilateral middle cerebral artery occlusion (MCAO) in 11 anesthetized baboons. To model early reperfusion, the clips were removed 6 h after occlusion. Successive measurements of regional CBF (rCBF), regional CMRO2 (rCMRO2), regional cerebral blood volume, and regional OEF (rOEF) were performed during the acute (up to 2 days) and chronic (> 15 days) stage. Late magnetic resonance imaging (MRI) scans (co-registered with PET) were obtained to identify infarction. Reversible MCAO produced an MRI-measurable infarction in 6 of 11 baboons; the others had no evidence of ischemic damage. Histological analysis confirmed the results of the MRI investigation but failed to show any evidence of cortical ischemic damage. The lesion was restricted to the head of the caudate nucleus, internal capsule, and putamen. The infarct volume obtained was 0.58 +/- 0.31 cm3. The infarcts were situated in the deep MCA territory, while the area of initially maximally increased OEF was within the cortical mantle. The mean absolute rCBF value in the infarct region of interest (ROI) was not significantly lower than in the highest-OEF ROI until 1-2 days post-MCAO. Cerebral metabolism in the deep MCA territory was always significantly lower than that of the cortical mantle; decreases in CMRO2 in the former region were evident as early as 1 h post-MCAO. In the cortical mantle, the rOEF was initially significantly higher than in the infarct-to-be zone. Subsequently, the OEF declined in both regions. The differences in the time course of changes in CMRO2 and OEF between these two regions, with the eventually infarcted area showing earlier metabolic degradation and in turn decline in OEF, presumably underlie their different final outcomes. In conclusion, following MCAO, the region that shows an early maximal increase in the OEF is both topographically and physiologically distinct from the region with final consolidated infarction if reperfusion is allowed at 6 h. This high OEF, although indicative of a threatened condition, is not an indicator of inescapable consolidated infarction and is thus a situation in which therapy could be envisaged. Whether or not it is at risk of infarction and thus constitutes one target for therapy remains to be seen.

Animals↗

PubMed1996

Apoptotic death in cortical neurons of mice subjected to focal ischemia.

Permanent focal cortical ischemia was induced in mice by electrocoagulation of the middle cerebral artery. At different time intervals after the injury, the volume of infarction was assessed together with an analysis of neuronal death. Morphological studies of ischemic brains and detection of nucleosomal DNA ladder within ipsilateral cortices might implicate a component of this neuronal loss to apoptosis as well as necrosis. Furthermore, we used the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling) procedure to detect in situ DNA fragmentation. The localization and the proportion of apoptotic cells in the ischemic mouse brain would indicate that apoptosis contributes largely to the cellular loss induced by cerebral ischemia.

Animals↗

PubMed1995

Galanin inhibits the vasodilatatory basalocortical cholinergic system in the anaesthetized rat.

In order to test the putative interaction between galanin and the vasodilatatory basalocortical cholinergic system, anaesthetized ventilated rats received a microinjection into the substantia innominata of 0.9% NaCl, 50 nmol carbachol, 50 nmol carbachol and 200 ng galanin, or 200 ng galanin. Cerebral blood flow (CBF) was measured with [14C]iodoantipyrine by the tissue sampling technique immediately following the intracerebral infusions. Under coinjection conditions, the flow increases observed after carbachol microinjection in the ipsilateral temporal and frontoparietal cortices were found to be significantly reduced (-37%, p < or = 0.02 and -25%, p < or = 0.05 respectively) compared with carbachol stimulated rats. The infusion of galanin by itself had no effect on CBF. These results demonstrate that galanin inhibits the vasodilatatory basalocortical cholinergic system and thus may possibly influence CBF by indirect mechanisms.

Acetylcholine↗