Dalia M. Araujo

Induction of immune system mediators in the hippocampal formation in Alzheimer's and Parkinson's diseases: Selective effects on specific interleukins and interleukin receptors

The present study determined whether molecules normally associated with immune signalling processes, specifically the lymphokines interleukin-1 beta, -2, -3 and -6, can be detected in the human hippocampal formation, and whether their levels are altered in neurodegenerative diseases such as Alzheimers and Parkinsons diseases. Interleukin-1 beta, -2, -3 and -6 were measured in post mortem tissues from 14 control neurologically normal subjects, 24 patients with Alzheimers disease and 17 patients with Parkinsons disease. In order to assess the extent of the cholinergic deficit in the Alzheimers disease brains, choline acetyltransferase activity in the hippocampal formation was first determined. In the Alzheimers disease tissues, choline acetyltransferase activity was significantly reduced (by 58%) compared to the control hippocampi, whereas that in the Parkinsons disease hippocampi was not significantly different from control. Using radioimmunoassays with antisera specific for the respective interleukin, marked increases in the content of immunoreactive interleukin-1 beta (99%), interleukin-2 (129%) and interleukin-3 (64%) could be detected in the Alzheimers, but not the Parkinsons disease hippocampi. Interleukin-6 levels were not significantly different in either group, compared to the control hippocampi. Since striking elevations in various interleukins were detected in the Alzheimers disease hippocampi, the possibility that concomitant alterations in interleukin receptor sites also occurred was investigated. Using radioligand binding to hippocampal membranes, low levels of interleukin binding were measured in the control hippocampi. In the Alzheimers tissues, significant elevations in [125I]interleukin-1 beta (by 65%) and [125I]interleukin-2 (by 69%) binding were noted. In contrast, [125I]interleukin-3 binding was not different in the Alzheimers disease compared to the control tissues. In the hippocampal formation of Parkinsons disease brains, only [125I]interleukin-2 binding was significantly increased (by 80%). In summary, the present results indicate that there is pronounced activation of immune system function, particularly specific immune mediators such as the interleukins, in the hippocampal formation in Alzheimers disease, and further suggest that stimulation of immune function may be an integral component of the pathological changes that occur in this disease.

Systemic interleukin-1β decreases brain-derived neurotrophic factor messenger RNA expression in the rat hippocampal formation

Brain-derived neurotrophic factor is selectively expressed at relatively high levels in the rat hippocampal formation (for review, see Ref. 12; see also Refs 8, 13, 19, 20, 27) where it is thought to be involved in mechanisms of neurodegeneration and/or neural protection related to the plasticity of hippocampal neurons. Functional responses to brain-derived neurotrophic factor appear to be mediated by a tyrosine receptor kinase B with the possible involvement of the p75 low-affinity nerve growth factor receptor protein. Among the many characteristics of Alzheimers disease is an upregulation of immune mediators in and around senile plaques in Alzheimers disease. Recently, interleukin-1 has been shown to be detrimental to the long-term survival of embryonic hippocampal neurons in culture. Thus, if the same occurs in vivo, it is possible that the accumulation of interleukin-1 in Alzheimers disease hippocampus may be responsible for altered hippocampal neuron synaptic plasticity. This may occur either by a direct action of interleukin-1 on hippocampal neurons or possibly indirectly by stimulating beta-amyloid production. Other indirect mechanisms may involve growth or survival factors such as the neurotrophin brain-derived neurotrophic factor which is thought to play an important role in the plastic responses of hippocampal neurons. A recent study showed that brain-derived neurotrophic factor mRNA is selectively decreased in the dentate gyrus in Alzheimers disease. The reason(s) for the decrease of brain-derived neurotrophic factor mRNA is not known, but one possibility may be associated with the enhanced expression of interleukin-1 in the hippocampus of Alzheimers disease patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Cholinergic regulation of hippocampal brain-derived neurotrophic factor mRNA expression: Evidence from lesion and chronic cholinergic drug treatment studies

Quantitative in situ hybridization and northern blot analysis techniques were used to determine the effects of removal of the cholinergic input on levels and topographical distribution of brain-derived neurotrophic factor mRNA in the hippocampus of adult rats. First, the effects of partial and full fimbrial transections, which result in partial and near-total cholinergic deafferentation respectively, were compared. Twenty-one days after partial unilateral fimbrial transections, there were significant decreases in brain-derived neurotrophic factor mRNA expression throughout the hippocampal formation. Decreased expression of brain-derived neurotrophic factor mRNA was evident in all areas of localization within the hippocampal formation. The decreases amounted to 22-36% reductions compared with unlesioned control animals. Brain-derived neurotrophic factor mRNA levels were decreased to a greater extent (50-69%) following full unilateral fimbrial transections. Quantitative northern blot analysis indicated that hippocampal BDNF mRNA was decreased by 29 and 68%, three weeks after partial or full unilateral fimbrial transections, respectively. The extent of the reductions in brain-derived neurotrophic factor mRNA levels correlated with reductions in acetylcholinesterase staining density and cholinergic terminal density determined by quantitative autoradiographic analysis of [3H]vesamicol binding sites. Second, we found that chronic treatment with atropine (20 mg/kg per day for 14 days) decreased (by 54%) brain-derived neurotrophic factor mRNA levels in all areas of localization within the hippocampus. In contrast, chronic treatment with nicotine (1.18 mg/kg per day for 14 days), a treatment known to desensitize nicotinic receptors, did not affect brain-derived neurotrophic factor mRNA expression in the hippocampal formation. The findings provide evidence for cholinergic muscarinic regulation of brain-derived neurotrophic factor mRNA expression in the adult rat hippocampal formation and they suggest the existence of a tonic stimulation of brain-derived neurotrophic factor synthesis by the cholinergic afferents.

BDNF and trkB mRNA expression in the hippocampal formation of aging rats.

Quantitative in situ hybridization and northern blot analysis techniques were used to determine the topographical distribution and levels of mRNA coding for brain-derived neurotrophic factor (BDNF) and the tyrosine receptor kinase (trkB) mRNA in the hippocampal formation of two strains of male rat during aging. Age did not change the prevalence or regional distribution patterns of BDNF or trkB mRNA in the hippocampal formation throughout the lifespan of male Sprague-Dawley rats. There also were no significant differences in the prevalence or topographical distribution patterns of trkB mRNA transcripts during aging. Northern blot analysis of hippocampal RNA from male Fischer 344 confirmed that neither BDNF mRNA nor trkB mRNA levels changed with age. These findings suggest that age-related neurodegenerative changes, including the atrophy of the cholinergic septo-hippocampal pathway, are not the result of changes in hippocampal BDNF or trkB mRNA expression. Moreover, the prevalence and distribution of synaptosomal-associated protein 25 (SNAP-25), a neuron-specific protein located in synaptic terminals and a putative marker of synaptic integrity, did not change with age. These findings indicate that altered hippocampal synaptic plasticity which occurs in the aged rat brain is not a reflection of changes in the expression of BDNF or trkB receptor mRNA.

Chronic intranigral administration of brain-derived neurotrophic factor produces striatal dopaminergic hypofunction in unlesioned adult rats and fails to attenuate the decline of striatal dopaminergic function following medial forebrain bundle transection

The present study determined the effects of chronic intranigral injections of recombinant human brain-derived neurotrophic factor (1 micrograms) every second day for 19 days on the functional capacity of dopaminergic neurons of the nigrostriatal pathway of unlesioned adult rats. In animals chronically treated with brain-derived neurotrophic factor, we observed amphetamine (5 mg/kg)-induced circling behavior directed toward the neurotrophin-injected side (33 turns/5 min). The behavioral asymmetry was paralleled by reductions of striatal [3H]dopamine uptake (27%), tyrosine hydroxylase activity (68%), dopamine content (36%) and [3H]mazindol binding site density (35%) on the same side as brain-derived neurotrophic factor treatment. While chronic injections of brain-derived neurotrophic factor produced a modest decrease in the number of tyrosine hydroxylase-positive cell bodies in the vicinity of the injection site, a similar reduction in cell number was observed in animals injected with a control protein, cytochrome c. However, in contrast to the animals treated with brain-derived neurotrophic factor, rats treated with the control protein showed no amphetamine-induced circling behavior, and there were no significant reductions in neurochemical parameters of striatal dopaminergic function. Lastly, we found that in brain-derived neutrophic factor-injected animals there was a 30% decrease of tyrosine hydroxylase messenger RNA levels in the ventral mesencephalon. We also determined the effects of brain-derived neurotrophic factor treatment on animals with transections of the medial forebrain bundle. Medial forebrain bundle-lesioned animals challenged with amphetamine circled (55 turns/5 min) ipsilateral to the lesioned side. The medial forebrain lesions decreased the following markers of striatal dopaminergic function: [3H]opamine uptake (65%), tyrosine hydroxylase activity (79%), dopamine content (80%) and [3H]mazindol binding site density (52%), induced a pronounced loss of tyrosine hydroxylase-positive cell bodies within the substantia nigra and also reduced tyrosine hydroxylase messenger RNA levels. Chronic intranigral brain-derived neurotrophic factor treatment did not attenuate nor did it exacerbate the medial forebrain bundle lesion-induced decreases of dopaminergic parameters in either the substantia nigra or striatum. The results of the present study indicate that chronic intranigral administration of brain-derived neurotrophic factor to normal adult rats induces a dopaminergic hypofunction in the striatum which is manifested behaviorally by amphetamine-induced rotations. The brain-derived neurotrophic factor-induced striatal function is not the result of significant cell loss at the levels of the substantia nigra, but seems to be related to brain-derived neurotrophic factor-induced down-regulation of dopaminergic-specific proteins.(ABSTRACT TRUNCATED AT 400 WORDS)

Distribution of [125I]nerve growth factor in the rat brain following a single intraventricular injection: Correlation with the topographical distribution of trk a messenger RNA-expressing cells

The present study determined the topographical distribution of [125I] nerve growth factor in rat brain at various time points following an intraventricular injection. In addition, we quantified the tissue content of nerve growth factor in various brain tissues following the injection. Autoradiographic analysis of the distribution of [125] nerve growth factor indicated that the neurotrophin is rapidly distributed within the entire ventricular system. However, penetration of nerve growth factor into the brain parenchyma was very limited. At early time points following an injection of nerve growth factor, there was an accumulation of label in the immediate vicinity of the lateral ventricle and third ventricle with predominant labeling around the septum, hypothalamus and cerebellum. By 24 h following nerve growth factor administration, there was discreet labeling of the lateral septum, medial septum, diagonal band, hypothalamus, olfactory tubercle and nucleus of the olfactory tract, and some label was present in the hippocampus and subiculum. Quantitative ELISA of nerve growth factor in brain tissues 1 h following the injection indicated a 446% and 133% increase over basal levels of nerve growth factor in the basal forebrain and hippocampus, respectively. At 24 h nerve growth factor levels measured in brain were not significantly different from endogenous basal levels as determined by ELISA, whereas there were high quantities of 125I present in the thyroid gland, suggesting that the administered [125I] nerve growth factor was rapidly degraded following the intraventricular injection. We observed a similar labeling pattern of the medial septum/diagonal band cholinergic cell body group 24 h following either an intraventricular or intrahippocampal injection of [125I] nerve growth factor. There was a good correlation between the [125I] nerve growth factor labeling pattern and the presence of trkA messenger RNA. This suggested that, at least in the septohippocampal pathway, nerve growth factor accumulated in a region which contained trkA nerve growth factor receptors. Thus, this study shows that after a single unilateral intraventricular injection of nerve growth factor into rat brain there is effective uptake by diagonal band/septal cells on both sides of the brain, and by cells whose positions correlate with the locations of cholinergic and trk A messenger RNA-expressing cells. Significant uptake was also observed in the hypothalamus and cerebellum. The very limited penetration and rapid degradation of intraventricularly administered nerve growth factor suggests that tissue penetration may be a limiting factor when attempting to influence brain neurons by exogenous neurotropic factors.

Effects of Chronic Basic Fibroblast Growth Factor Administration to Rats with Partial Flmbrial Transections on Presynaptic Cholinergic Parameters and Muscarinic Receptors in the Hippocampus: Comparison with Nerve Growth Factor

Abstract: The present study compares the effects of chronic administration of basic fibroblast growth factor (bFGF) and nerve growth factor (NGF) on various hippocampal cholinergic parameters in rats with partial unilateral fimbrial transections. Lesions resulted in marked reductions of several presynaptic cholinergic parameters: choline acetyltransferase (ChAT) activity (by 50%), [3H]‐acetylcholine ([3H]ACh) synthesis (by 59%), basal and ve‐ratridine (1 μM)‐evoked [3H]ACh release (by 44 and 57%, respectively), and [3H]vesamicol binding site densities (by 35%). In addition, [3H]AF‐DX 116/muscarinic M2 binding site densities were also modestly decreased (by 23%). In contrast, [3H]pirenzepine/muscarinic M1 and [3H]AF‐DX 384/muscarinic M2/M4 binding site densities were not altered by the lesions, nor were they affected by any of the treatments. Intracerebroventricular administration of bFGF (10 ng, every other day, for 21 days) partially prevented the lesion‐induced deficit in hippocampal ChAT activity, an effect that was not markedly different from that measured in the NGF‐treated (1 μg intracerebroventricularly, every other day, for 21 days) rats. In rats treated with a combination of bFGF and NGF, ChAT activity was not different from that in rats treated with the individual factors alone. In contrast, the lesion‐induced deficits in the other cholinergic parameters were not attenuated by bFGF treatment, although they were at least partially prevented by NGF administration. To determine whether higher concentrations of bFGF are necessary to affect cholinergic parameters other than hippocampal ChAT activity, rats were treated with 1 μg (every other day, 21 days) of the growth factor. In this group of rats, detrimental effects of bFGF, manifested by an increased death rate (46%), and marked reductions in body weight of the survivors, were observed. In addition, this concentration of bFGF appeared to exacerbate the lesion‐induced reduction in [3H]ACh synthesis by hippocampal slices; [3H]ACh synthesis in lesioned hippocampi represented 36 and 52% of that in contralateral unlesioned hippocampi for the bFGF‐treated and control groups, respectively. In conclusion, although bFGF administration attenuates the deficit in hippocampal ChAT activity induced by partial fimbrial transections, this does not appear to translate into enhanced functional capacity of the cholinergic terminals. This is clearly in contrast to NGF, which enhances not only hippocampal ChAT activity, but also other parameters indicative of increased function in the cholinergic terminals.

Regulation of hippocampal muscarinic receptor function by chronic nerve growth factor treatment in adult rats with fimbrial transections.

Effects of chronic intraventricular administration of recombinant human nerve growth factor on hippocampal muscarinic receptor densities and muscarinic receptor-linked second messenger systems were determined in adult rats 21 days following partial or full unilateral fimbrial transections. First, autoradiographic analysis of muscarinic receptors was carried out using [3H]quinuclidinyl benzilate for total muscarinic receptors, [3H]pirenzepine for M1 receptors and [3H]AF-DX 384 for M2 receptors. Partial fimbrial transections did not significantly alter the density of these muscarinic receptor populations in the dorsal or ventral hippocampus and there was no effect of chronic (1 micrograms every other day, 21 days) recombinant human nerve growth factor treatment. In contrast, in animals receiving full fimbrial transections which by themselves did not alter muscarinic receptor density, recombinant human nerve growth factor treatment increased the density of [3H]quinuclidinyl benzilate binding sites, M1 receptors, and M2 receptors by approximately 40% in the CA1 region. Secondly, we determined the effect of chronic recombinant human nerve growth factor treatment on muscarinic receptor-mediated second messenger production in rats with either partial or full unilateral fimbrial transections. In partially fimbriectomized rats, oxotremorine-induced inositol triphosphate production by hippocampal slices was increased by 81% on the lesioned side of animals treated with a control protein. This lesion-induced supersensitivity of M1 muscarinic receptor function was prevented by chronic recombinant human nerve growth factor treatment. In recombinant human nerve growth factor-treated animals, inositol triphosphate production was similar to values on unlesioned control sides. The muscarinic receptor-mediated increase in cyclic GMP levels was not altered by fimbrial transections or recombinant human nerve growth factor treatment. In animals with full unilateral fimbrial transections, oxotremorine-induced inositol triphosphate production was increased by 99% on the lesioned side of animals treated with a control protein and treatment with recombinant human nerve growth factor did not alter this denervation-induced supersensitivity of muscarinic receptor transduction signal. Chronic recombinant human nerve growth factor treatment did not affect the levels of inositol triphosphate on the contralateral unlesioned side of either partial or full fimbriectomized animals. Earlier studies indicate that chronic nerve growth factor treatment increases the presynaptic function of hippocampal cholinergic neurons surviving partial fimbrial transections. The findings of the present study indicate that these presynaptic effects translate into functional changes at the level of postsynaptic muscarinic receptors in the hippocampus.(ABSTRACT TRUNCATED AT 400 WORDS)

NGF suppression of weight gain in adult female rats correlates with decreased hypothalamic cholecystokinin levels

Effects of chronic intraventricular administration of nerve growth factor (NGF, 1 microgram qod for 21 days) on weight gain, hypothalamic neuropeptide levels and choline acetyltransferase (ChAT) activity were determined in adult female Wistar rats. Rats chronically treated with cytochrome c (cc) gained 163 g over the 21 day treatment schedule, whereas NGF-treated rats only gained 110 g. Thus, NGF-treated rats gained 53 g less; this change in weight gain is equivalent to approximately a 20% decrease of total weight gain compared to the cc-treated control rats. Chronic NGF treatment significantly decreased hypothalamic cholecystokinin (CCK) levels by 24% (P = 0.0070), but did not alter either hypothalamic neuropeptide Y (NPY) or bombesin (BOMB) levels (98% and 105% of cc-treated control levels, respectively). In addition, chronic NGF treatment did not significantly alter hypothalamic ChAT activity (95% of cc-treated control rats). The results of the present study suggest that NGF-induced decreases in weight gain are not the result of alterations of hypothalamic cholinergic function. However, it is possible that NGF-induced alterations of hypothalamic CCK synthesis and release may be involved in the NGF-induced decrease in weight gain.

Effects of chronic nerve growth factor treatment on hippocampal [3H]cytisine/ nicotinic binding sites and presynaptic nicotinic receptor function following fimbrial transections

Recent studies with nerve growth factor (NGF) have identified the pharmacological actions of this neurotrophin in a variety of animal models that mimic some of the neurotransmitter deficits that occur in Alzheimers disease (AD, for reviews see Refs 7, 15, 17, 19). Based upon extensive pharmacological studies, NGF has been characterized as a crucial maintenance factor for adult cholinergic neurons of the septo-hippocampal and basalo-cortical pathways. Among the reported actions of NGF is an attenuation of lesion-induced decrements in presynaptic and postsynaptic cholinergic markers and functions in the hippocampal formation. Thus, in studies that used partial fimbriectomies to parallel the cholinergic neurodegeneration that occurs in AD, intraventricularly administered nerve growth factor prevented the loss of choline acetyltransferase (ChAT) and acetylcholinesterase immunoreactivity in the septum and increased a variety of presynaptic cholinergic markers involved in the synthesis, storage and release of the neurotransmitter acetylcholine (for reviews see Refs 7, 17, 19). More specifically, chronic NGF treatment attenuates lesion-induced reductions in hippocampal ChAT activity and high-affinity choline uptake, the end-result of which is an enhanced capacity to synthesize acetylcholine. This increased acetylcholine synthesis, in turn, appears to translate directly into augmented vesicular storage and release of the neurotransmitter. For instance, not only does NGF treatment reverse lesion-induced reductions in maximal binding densities of the acetylcholine vesicular transport marker [3H]vesamicol, but it also enhances acetylcholine release and turnover rate. NGF treatment also appears to restore the sensitivity of postsynaptic muscarinic receptors to agonist-induced stimulation following partial fimbriectomies.(ABSTRACT TRUNCATED AT 250 WORDS)