AMPA receptor regulation and LTP in the hippocampus of young and aged apolipoprotein E-deficient mice.

In the present study, modulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors by phosphatidylserine (PS) and synaptic plasticity were investigated in the hippocampus of young (4-month-old) and aged (18-month-old) apolipoprotein E (apoE)-deficient mice. Qualitative as well as quantitative analysis of brain sections in both young and aged apoE-deficient mice did not reveal any substantial changes of AMPA receptor binding in the various hippocampal regions, compared to age-matched controls. Nevertheless, enhancement of AMPA receptor binding elicited by PS treatment was found to be abolished in most hippocampal regions of young apoE-deficient mice, while modulation of AMPA receptors by this phospholipid was not significantly altered in the hippocampal formation of aged apoE-deficient animals. At the electrophysiological level, long-term potentiation (LTP) induced by theta burst stimulation was lower in area CA1 of the hippocampus of young, but not aged, apoE-deficient mice compared to age-matched controls. These results confirm that apoE is important for AMPA receptor regulation and LTP expression in the hippocampal formation. However, the presence of LTP in aged apoE-deficient animals, together with apparent recovery of the PS action on AMPA receptors, suggests that aged apoE-knockout mice possess compensatory mechanisms that reduce biochemical and electrophysiological alterations of glutamatergic neurons.

Impact of apoE deficiency on oxidative insults and antioxidant levels in the brain.

Apolipoprotein E (apoE) is a lipid transport molecule, which has been linked to the pathogenesis of Alzheimer's disease. Recently we have demonstrated that the oxidative insults in hippocampus from AD patients were dependent on the apoE genotype. Interestingly, apoE protein concentration in hippocampus follows a genotype-dependent gradient with the lowest level occurring in varepsilon4 allele carrier. We raised the possibility that, in the hippocampus, the apoE level affects the oxidant/antioxidant balance. Here, we have examined in the apoE-deficient mouse the oxidant/antioxidant status in hippocampus and in frontal cortex from APOE-KO and wild-type mice at 3 and 13 months. We provided evidence that, in the hippocampus, the absence of apoE has a clear impact on the oxidant/antioxidant status. Endogenous level of thiobarbituric acid-reactive substances (TBARS) was found to be markedly elevated whereas level of alpha-tocopherol was decreased in APOE-deficient mice at 3 and 13 months. Superoxide dismutase activities were also lower in APOE-deficient mice at 13 months. Taken together, these data indicate that the steady state level of apoE may influence, to a certain extent, the balance between oxidants and antioxidants in hippocampus.

Phospholipase A2 activity is decreased selectively in the hippocampus of aged apolipoprotein E deficient mice.

Because apolipoprotein E (ApoE) deficient mice have cognitive deficits (Neurosci. Lett., 199 (1995) 1-4; Neuroscience, 92 (1999) 1273-1286; Brain Res., 752 (1997) 189-196) that may involve decreased phospholipase A(2) (PLA(2)) activity (Neuroscience, 92 (1999) 1273-1286), striatal, hippocampal, and parieto-temporal PLA(2) activity was measured in cytosol from 3 and 20-month-old ApoE deficient and control mice. Samples were homogenized and cytosol prepared by ultracentrifugation. PLA(2) activity in each cytosolic fraction was measured in triplicate using a continuous fluorometric assay (J. Neurosci. Methods (2000) in press). In 3-month-old animals, there was a trend for decreased hippocampal PLA(2) activity between groups. In 20-month-old animals, hippocampal PLA(2) activity was significantly (P=0.0304) decreased nearly 20% in ApoE deficient mice as compared to age-matched control mice. No differences were found in other brain regions, although activity in the striatal samples were nearly 65% less than in the other two regions.

Cholinergic systems and long-term potentiation in memory-impaired apolipoprotein E-deficient mice.

Impairments in cholinergic neurotransmitter systems of the basal forebrain are a hallmark of Alzheimer's disease pathophysiology. The presence of the epsilon4 allele of apolipoprotein E was recently implicated as a major risk factor in both familial and sporadic Alzheimer's disease. The present study examined the integrity of cholinergic and non-cholinergic systems in apolipoprotein E-deficient, memory-impaired mice. Choline acetyltransferase activity, hippocampal acetylcholine release, nicotinic and muscarinic (M1 and M2) receptor binding sites and acetylcholinesterase cell or terminal density showed no signs of alteration in either three-month or 9.5-month-old apolipoprotein E-deficient mice compared to controls. In contrast, long-term potentiation was found to be markedly reduced in these mice, but increases in the strength of stimulation induced the same level of long-term potentiation as that observed in controls. These alterations did not appear to be the consequence of modifications in the binding properties of glutamatergic receptors (N-methyl-D-aspartate and [RS]-alpha-amino-3-hydroxy-5-methylisoxazole propionic acid) but from defective regulation of the (RS)-alpha-amino-3-hydroxy-5-methylisoxazole propionic acid receptor by phospholipase A2 activity. These results support the notion that apolipoprotein E plays a fundamental role in neuronal plasticity, which could in turn affect cognitive performance through imbalances in extra- and intracellular lipid homeostasis.

The neurobiology of apolipoproteins and their receptors in the CNS and Alzheimer's disease.

The importance of apolipoproteins in the central nervous system became increasingly clear with the association in 1993 of the epsilon4 allele of apolipoprotein E with familial and sporadic late-onset Alzheimer's disease. Apolipoprotein E is a ligand for several receptors, most of which are found to some extent in the brain. This review summarizes the various apolipoproteins and lipoprotein receptors found in the brain. A growing body of evidence now implicates irregular lipoprotein metabolism in several neurodegenerative disorders. We then focus on research linking apolipoprotein E and Alzheimer's disease, from clinical studies to biochemical models, which may explain some of the complex neurobiology of this disorder.

Synaptic mechanisms and calcium binding proteins in the aged rat brain.

Synaptic mechanisms were studied ex vivo in the aged rat hippocampus, using a slice preparation and intracellular electrophysiological recordings of the CA1 pyramidal neurons. A dramatic depression of the slow cholinergic excitatory postsynaptic potential (EPSP) and of the slow, GABAB-mediated inhibitory postsynaptic potential (IPSP) were observed. These age-related changes were consistently found in three different strains of rats. The mechanisms involve 1) changes in the properties of the postsynaptic muscarinic receptors, and possibly in acetylcholine release (for the postsynaptic muscarinic receptors, and possbily in acetylcholine release (for the cholinergic EPSP), and 2) alterations in the presynaptic GABAergic interneurons, as shown by a loss in calbindin immunoreactivity (for the GABAergic IPSP). The immunoreactivity for three calcium binding proteins (calbindin, parvalbumin and calretinin) was studied in the aged rat brain. Immunoreactivity for calbindin was dramatically reduced in the pyramidal neurons of the CA1 field and in a subpopulation of interneurons in the hippocampus. Immunoreactivity for parvalbumin was reduced in the medial septal area, and in the cingulate cortex, whereas no change was observed for calretinin. These age-related alterations could 1) modify the functions of the hippocampal networks, and possibly contribute to the age-related cognitive deficits, and 2) compromise intraneuronal calcium buffering, and thus make neurons more vulnerable to toxic insults.

Calretinin-containing pathways in the rat forebrain.

The anatomy of pathways containing the calcium binding protein calretinin was investigated in the forebrain of the rat, using a combination of immunohistochemical and retrograde tract tracing techniques. Numerous well identified pathways do contain calretinin, whereas others do not. Pathways arising from the substantia nigra/ventral tegmental area, the dorsal raphe, the lateral mammillary nucleus, the supramammillary nucleus, the triangular septal and septo-fimbrial nuclei, several thalamic nuclei, the parabrachial nucleus, the peripeduncular nucleus, the medial amygdala contain at least some calretinin. The proportion of projection neurons containing calretinin ranged from 2% (dorsal raphe to caudate) to about 75% (triangular septal nucleus to habenula, medial amygdala to the ventromedial hypothalamus). More than 50% of the nigro-striatal neurons contain calretinin immunoreactivity. In contrast, other pathways do not contain any calretinin immunoreactivity (for instance the pathways arising from cerebral cortex, locus coeruleus, cholinergic forebrain nuclei), although calretinin may be present in local neurons in these structures. The present study demonstrates that calretinin is not associated specifically with projection neurons or local neurons, identified transmitter systems or functionally related pathways in the forebrain of the rat.

Age-related changes in parvalbumin- and GABA-immunoreactive cells in the rat septum.

The calcium binding protein parvalbumin is present in GABAergic neurons of the medial septum-diagonal band of Broca (MS-DBB) region that project to the hippocampal formation. We examined the distribution pattern, the number, and the morphological features of the parvalbumin-containing cells (parv+) in the MS-DBB region of 2- to 3-, 8- to 9-, 15- to 16-, and 26- to 27-month-old Sprague-Dawley rats. A significant reduction in the number of parv+ cells was observed as a function of age. The mean somal area of the parv+ cells was significantly reduced in the 26- to 27-month-old rats. A significant reduction in the number of parv+ cells was also observed in the 26- to 27-month-old rats in the cingulate cortex, but not in the striatum or the hippocampal formation. No significant age-related changes were observed in the number of the GABA-immunoreactive cells in the MS-DBB region nor in the cingulate cortex. In conclusion, there is an age-related decrease in the number of parv+ cells, with no change in the number of GABA-immunoreactive cells in the MS-DBB region of the rat. Because GABA and parvalbumin are colocalized in the MS-DBB neurons, the results suggest that the level of parvalbumin is decreased, but that the cells are not lost.

Loss of calbindin-immunoreactivity in CA1 hippocampal stratum radiatum and stratum lacunosum-moleculare interneurons in the aged rat.

Alterations in hippocampal circuitry may underly age-related learning and memory impairment. We showed in a previous study that the GABAB-mediated slow inhibitory postsynaptic potential (IPSP) induced in CA1 pyramidal neurons by electrical stimulation of stratum radiatum, is depressed in the hippocampus of the aged rat. This could be due to alterations in GABAergic interneuron functions. We report in this study that the number of hippocampal calbindin-immunoreactive (CaBP-IR) GABAergic interneurons is decreased in the aged rat. The mean number of CaBP-IR interneurons per slice decreases by 50% in the aged rat. The most severe loss was observed in the stratum radiatum of CA1 (78%), with a less consistent loss of immunoreactivity in CA3 (35%). In contrast, the mean number of interneurons containing parvalbumin (PV), was not significantly decreased in the aged rat. Our results show a loss of CaBP immunoreactivity in a population of GABAergic interneurons, which might be related to an altered function of these interneurons and consequently of GABAergic synaptic transmission in the aged rat. In contrast, PV immunoreactivity in interneurons located close to the pyramidal layer does not decrease in the hippocampus of the aged rat.

The age-related increase in galanin binding sites in the rat brain correlates with behavioral impairment.

The regional distribution of [125I]galanin specific binding sites was determined in young (three- to four-month-old), 14-15-month-old and aged (26-27-month-old) male Sprague-Dawley rats, previously tested for their performances in the Morris water-maze task, using the radioautographic method on brain sections. A significant increase in specific binding was observed in piriform and entorhinal cortex, ventral subiculum, and dorsal dentate gyrus in the aged rats, whereas no significant changes were observed in dorsal subiculum, amygdala, septal area and various subcortical structures. The area-specific regional increase in specific binding density in aged rats was significantly correlated with the impairment of the behavioral performance in the Morris water-maze task. The change in [125I]galanin specific binding was a result of an increase in the number of galanin binding sites, but not of an increase in affinity.

Age-related increase in galanin-binding sites in the rat brain: correlation with behavioral impairment.

The regional distribution of 125I-galanin specific binding sites was determined by radioautography on brain sections in young (3- to 4-month-old) and aged (26- to 27-month-old) male Sprague-Dawley rats, previously tested for their performances in the Morris water maze task. In aged rats, a significant increase in specific binding was observed in piriform, perirhinal and entorhinal cortex, the CA1 field of the ventral hippocampus, ventral subiculum, and dorsal dentate gyrus, whereas no significant change was observed in the ventral dentate gyrus, the dorsal subiculum, the CA3 field of the hippocampus, the amygdala or the septal area. The area-specific regional increase in specific binding density in aged rats was significantly correlated with the impairment of their behavioral performances in the Morris water maze task. The change in 125I-galanin specific binding in the aged rats was a result of an increase in the number of galanin-binding sites, without change in affinity.