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Cholinergic innervation of the primate hippocampal formation. I. Distribution of choline acetyltransferase immunoreactivity in the Macaca fascicularis and Macaca mulatta monkeys.

The cholinergic innervation of the hippocampal formation of Macaca fascicularis (cynomolgus) and Macaca mulatta (rhesus) monkeys was investigated by immunohistochemical procedures using a monoclonal antibody directed against choline acetyltransferase. The distribution of choline acetyltransferase in the monkey demonstrated both similarities and differences with the staining patterns observed in the rat or with acetylcholinesterase in the monkey. While both of these latter preparations demonstrated labeled cells, for example, no choline acetyltransferase labeled neurons were observed in the monkey hippocampal formation. Choline acetyltransferase activity was restricted to fibers which varied in thickness and number of varicosities and in their regional and laminar distribution. The highest densities of labeled fibers were observed in the uncal portion of the hippocampus, in the parasubiculum, and in the entorhinal cortex; the lowest densities of labeled fibers were observed in CA1 and in midrostrocaudal levels of the dentate gyrus. In the dentate gyrus, immunoreactive fibers were densely distributed in the molecular layer and in an infragranular plexus. One of the few striking noticeable interspecies differences was observed in the dentate gyrus. In the rhesus monkey, labeled fibers in the molecular layer were divided into a superficial denser and an inner lighter lamina, whereas in M. fascicularis, the cholinergic fibers were distributed more homogeneously throughout the molecular layer. In the hippocampus proper, there was a progressive decrease in the density of ChAT-immunoreactive fibers from CA3/CA2 into CA1. The subiculum also demonstrated modest labeling which was nonetheless higher than in CA1; the border of these fields demonstrated increased fiber labeling. The density of choline acetyltransferase staining was high in the presubiculum and parasubiculum. In the entorhinal cortex, a relatively clear boundary was observed between the more heavily stained superficial layers (I, II, and III) and the more weakly labeled deep layers (V and VI), especially in the intermediate and caudal fields. A transverse decreasing gradient was observed with the densest plexus of cholinergic fibers found in the medially situated olfactory field of the entorhinal cortex and the lowest density in the laterally located caudal and lateral fields.

Animals↗

Phenytoin, phenobarbital, and midazolam fail to stop status epilepticus-like activity induced by low magnesium in rat entorhinal slices, but can prevent its development.

OBJECTIVES: It was shown previously that low-Mg2+-induced epileptiform activity in rat entorhinal cortex slices changes with time from a pattern of serial seizure-like events (SLEs) to a state of continuously recurring epileptiform activity. Valproic acid blocked the early SLEs but not the late activity. It was proposed that the late activity is a model for pharmacoresistant status epilepticus since it was also refractory to phenytoin, carbamazepine, phenobarbital, and midazolam. In the present study, it is demonstrated that phenytoin (50 microM, n=6), phenobarbital (150 microM, n=7), and midazolam (50 microM, n=5) were able to block the early SLEs but not the late activity at the same concentrations. Carbamazepine (50 microM) reduced the duration of the SLEs from 21 +/-5 s to 4+/-3 s (P<0.01), the interictal interval from 123+/-27 s to 27+/-19 s (P<0.01), the SLE-associated rise of [K+]o from 7.7+/-0.5 mM to 5.7+/-0.8 mM (n=4, P<0.05), and the spread of the SLE between entorhinal cortex and neocortex from 4.0+/-0.6 s to 0.8+/-0.1 s (n=4, P<0.05). Lower concentrations of phenytoin (5 and 10 microM, n=5), carbamazepine (10 microM, n =6), and phenobarbital (50 microM, n = 4) had no effect. In conclusion, the hypothesis is supported that low-Mg2+-induced epileptiform activity in rat entorhinal cortex is an in vitro model for the transition from pharmacosensitive to pharmacoresistant status epilepticus.

Animals↗

Exposure to trimethyltin significantly enhances acetylcholinesterase staining in the rat dentate gyrus.

Trimethyltin (TMT) is known to produce substantial damage to the hippocampal formation. It also destroys neurons within the entorhinal cortex, thereby causing degeneration of perforant path afferents that terminate in the outer molecular layer (OML) of the dentate gyrus. Surgical destruction of the entorhinal cortex also causes the perforant path to degenerate. This leads to reactive synaptogenesis (axonal sprouting) of septal afferents to the dentate gyrus. The purpose of the present study was to determine whether administration of 6 mg/kg of TMT by gavage to rats would cause axonal sprouting within the septo-dentate projection. A histochemical stain for acetylcholinesterase (AChE) was used. Compared to control subjects rats given TMT exhibited significantly denser AChE staining in the dentate OML. This is putative indication of reactive synaptogenesis within the cholinergic projection to this layer of the dentate and is somewhat surprising because other neurotoxins, such as lead and ethanol, that affect neurons within the hippocampal formation reduce the capacity for reactive synaptogenesis in response to lesions of the entorhinal cortex.

Acetylcholinesterase↗

Staging of Alzheimer-related cortical destruction.

The gradual intraneuronal accumulation of an insoluble fibrous material which partly consists of abnormally phosphorylated tau protein (neurofibrillary change) represents an important neuropathological hallmark of Alzheimer's disease. Neurofibrillary tangles and neuropil threads formed from this material develop in only a few types of cortical pyramidal cells. The first changes are seen in the entorhinal cortex. The destructive process then spreads into the hippocampal formation and eventually encroaches upon the isocortex. This sequence of events permits the distinction of six stages with a progressive increase in the severity of cortical destruction. The entorhinal region serves as an important interface between the isocortex and hippocampus. This interface function is markedly impaired due to the early deterioration of the entorhinal cortex. Severe entorhinal involvement is considered to represent the morphological counterpart of clinically incipient Alzheimer's disease. Similar changes are found in mentally impaired individuals suffering from Parkinson's disease or progressive supranuclear palsy.

Aged↗

Calretinin immunoreactivity in the monkey hippocampal formation--I. Light and electron microscopic characteristics and co-localization with other calcium-binding proteins.

Calretinin-containing neurons were visualized by immunocytochemistry in the monkey hippocampal formation, subicular complex, and entorhinal cortex. Calretinin-immunoreactivity was present exclusively in non-granule cells of the dentate gyrus and in non-pyramidal cells of Ammon's horn, subiculum and entorhinal cortex. Most frequently, calretinin-positive neurons were found at the hilar border of the dentate granule cell layer and in the stratum radiatum of CA1-3 areas. In the subicular complex, immunoreactive neurons were evenly distributed in all layers, whereas in the entorhinal cortex, they were accumulated in external layers above the lamina dissecans. Distinct bands of calretinin-positive fibers occupied the supragranular zone of the molecular layer in dentate gyrus, the pyramidal cell layer of the CA2 area in Ammon's horn and the upper two layers of presubiculum. The majority of calretinin-immunoreactive neurons were small, bipolar or fusiform neurons with a dendritic tree oriented parallel to the dendrites of principal cells (granule cells in dentate gyrus and pyramidal neurons elsewhere). Dendrites were smooth or sparsely spiny, displaying small spines of conventional type. Co-existence studies showed that these neurons were completely devoid of other calcium-binding proteins, parvalbumin and calbindin. Electron microscopic analysis revealed somata of immunoreactive neurons which contained a large nucleus and a small cytoplasmic rim, which contained only few organelles. The nucleus displayed deep infoldings and intranuclear rods. Input synapses of immunoreactive neurons were rare both on somata and dendrites and large surface areas were frequently apposed by glial processes. This was very prominent in the dentate gyrus and Ammon's horn. Axons of calretinin-positive neurons were thin, arborized in all layers and had small varicosities. Their terminals formed symmetric synaptic contacts mainly with dendrites and less frequently with somata of principal cells. Axon terminals of calretinin-immunoreactive fiber bundles in the supragranular layer, as well as in the pyramidal layer of the CA2 area, formed asymmetric synaptic contacts with dendritic shafts. In addition, they established asymmetric axospinous and axosomatic synaptic contacts with granule cells of the dentate gyrus. In the presubiculum, the calretinin-positive axon bundle included a large number of immunoreactive myelinated axons, as well as axon terminals. The characteristic location and features of synapses suggests that these fibers derive from extra-hippocampal afferents (Nitsch, R. and Leranth C. (1993) Neuroscience 55, 797-812) and not from the calretinin-immunoreactive neurons of the hippocampal formation.

Animals↗

Memory processing by the limbic system: role of specific neurotransmitter systems.

Experiments using localized infusions into selected brain structures of agonists and antagonists of various synaptic receptors, given before or after behavioral training, have led to the following conclusions: (1) Memory is processed shortly after training in the amygdala, medial septum and hippocampus by glutamatergic NMDA and AMPA receptors activated in that sequence. Cholinergic muscarinic receptors are activated concurrently with the former. GABAA receptors modulated by brain benzodiazepines and by beta-noradrenergic receptors inhibit the process. (2) The sequential involvement of NMDA and AMPA receptors suggests that long-term potentiation (LTP) of the synapses activated by the learning experiences in the hippocampus and/or amygdala and medial septum is the crucial event. Expression of this LTP at the time of testing is necessary for retrieval: AMPA receptor blockade in the hippocampus and amygdala at the time of testing hinders retrieval. This suggests that the LTP underlies the memory process itself. (3) The amygdala, medial septum and hippocampus mediate different types of memory and/or different components of memories. The entorhinal cortex, through mechanisms that require intact NMDA receptors and are inhibited by GABAA receptors, intervenes in post-training memory processing 90-180 min after the other limbic regions. The entorhinal cortex integrates consecutively acquired memories; this role could be maintained by the LTP that is generated after training in the amygdala, hippocampus and medial septum. Post-training intervention of the entorhinal cortex does not occur if this region is inhibited at the time of training.

Animals↗

The effect of hypoxia on the epileptiform activities induced by magnesium-free medium in rat brain slices.

In order to understand the mechanisms underlying the seizure generation, the present study has investigated the effect of hypoxia on the transition between seizure and interictal bursting. Bathing rat brain slices of the hippocampus and entorhinal cortex in magnesium-free medium elicits electrographic seizures. However, they are eventually replaced by the interictal bursts. It has previously been shown that the interictal bursts, arising in the hippocampal area CA3, are propagated to and disrupt the seizure generation in the entorhinal cortex. In this report we demonstrate that hypoxia promotes the seizure reappearance in the entorhinal cortex by suppressing the interictal bursts in CA3.

Animals↗

Low glial numbers in the amygdala in major depressive disorder.

BACKGROUND: Functional imaging studies implicate the prefrontal cortex and amygdala in major depressive disorder and bipolar disorder, and glial decreases have been reported in the prefrontal cortex. Here, glia and neurons were counted in the amygdala and entorhinal cortex in major depressive disorder, bipolar disorder, and control cases. METHODS: Tissue blocks from major depressive disorder (7), bipolar disorder (10), and control (12) cases, equally divided between right and left, were cut into 50 microm sections and stained with the Nissl method. One major depressive disorder and all but two bipolar disorder cases had been treated with lithium or valproate. Neurons and glia were counted using stereological methods. RESULTS: Glial density and the glia/neuron ratio were substantially reduced in the amygdala in major depressive disorder cases. The reduction was mainly accounted for by counts in the left hemisphere. No change was found in neurons. Average glia measures were not reduced in bipolar disorder cases; however, bipolar disorder cases not treated with lithium or valproate had significant glial reduction. Similar but smaller changes were found in the entorhinal cortex. CONCLUSIONS: Glia are reduced in the amygdala in major depressive disorder, especially on the left side. The results suggest that lithium and valproate may moderate the glial reduction.

Amygdala↗

Theta-modulated feedforward network generates rate and phase coded firing in the entorhino-hippocampal system.

Principal cells of the hippocampus and of its only cortical input region, the entorhinal cortex exhibit place specific activity in the freely moving rat. While entorhinal cells have widely tuned place fields, hippocampal place fields are more localized and determine not only the rate but also the timing of place cell spikes. Several models have successfully attempted to explain this fine tuning making use of intrahippocampal attractor network dynamics provided by the recurrent collaterals of hippocampal area CA3. Recent experimental evidence shows that CA1 place cells preserve their tuning curves even in the absence of input from CA3. We propose a model in which entorhinal and hippocampal pyramidal cell populations are only connected via feedforward connections. Synaptic transmission in our sytem is gated by a class of interneurons inhibiting specifically the entorhino-hippocampal pathway. Theta rhythm modulates the activity of each component. Our results show that rhythmic shunting inhibition endows entorhinal cells with a novel type of temporal code conveyed by the phase jitter of individual spikes. This converts coarsely tuned place-specific activity in the entorhinal cortex to velocity-dependent postsynaptic excitation and, thus, provides hippocampal place cells with an input that has recently been proposed to account for their rate and phase coded firing. Hippocampal place fields are generated through this mechanism and also shown to be robust against variations in the level of tonic inhibition.

Action Potentials↗

The role of the perforant pathway as a trophic factor for neurotransmission in the rat dentate gyrus.

Changes in electrophysiological function in the hilar associational pathway terminating on dentate granule cells in the rat hippocampal formation were studied following unilateral entorhinal cortex lesions. In rats lesioned as pups (postnatal day 4 [PN 4]) or as adults (PN 60) there was a profound loss of paired-pulse inhibition at 30 days postlesion. Inhibition was unaffected at 10 days postlesion. Entorhinal cortex lesions did not affect population spike amplitude, population excitatory postsynaptic potentials slopes, or long-term potentiation compared to the unlesioned hemisphere. The presence of a complete hippocampal commissurotomy had no effect on excitatory or inhibitory parameters. Laminar analyses of extracellular field potentials from animals lesioned as adults revealed an expansion of functional synapses outward into the dentate molecular layer. This expansion was complete by 10 postlesion days. The changes observed with laminar analyses were not contemporaneous with the changes in paired-pulse inhibition. The loss of inhibition in the hilar associational pathway of entorhinal cortex-lesioned animals thus implies a change in local circuit function rather than an effect from sprouted associational fibers directly onto granule cells. The lack of inhibition in the associational pathway in lesioned animals was not due to a failure of local circuit inhibitory function to develop, since the same findings were obtained when lesions were made neonatally or as adults. Rather, the authors suggest that the present findings arise because of the formation of functional, recurrent, excitatory mossy fiber collateral synapses following entorhinal cortex lesions.

Afferent Pathways↗

Quantitative evaluation of neurotrophin and trk mRNA expression in visual and limbic areas along the occipito-temporo-hippocampal pathway in adult macaque monkeys.

The neurotrophins have been implicated in shaping and remodeling the connectivity of neural circuits. To explore the role of neurotrophins and their receptors, Trks, in cortical neural circuits of adult macaque monkeys, we determined mRNA expression levels of neurotrophins and Trk receptors in various visual and limbic areas along the occipito-temporo-hippocampal pathway by using a quantitative reverse-transcription polymerase chain reaction technique. The expression level of brain-derived neurotrophic factor (BDNF) mRNA was lowest in the primary visual cortex (V1), moderate in the temporal visual association area, and highest in the hippocampus. The expression levels of trkB mRNA isoforms, the full-length form that encodes a receptor tyrosine kinase and the truncated form that encodes a noncatalytic receptor, were also low in V1, moderate in the visual association area, and high in the entorhinal cortex. However, in contrast to their ligand BDNF, the expression levels of both trkB isoforms in the hippocampus were significantly lower than those in the entorhinal cortex. NT-3 mRNA was detectable only in the hippocampus and the entorhinal cortex, whereas both the full-length and the truncated forms of trkC mRNA were widely distributed throughout the neocortex and the limbic cortex. The expression levels of NGF and trkA mRNAs in these cortical areas were too low to determine quantitatively. The present findings suggest that, among neurotrophin/Trk signaling systems, the BDNF/TrkB-mediated signal most likely contributes to stabilization, remodeling, or both, of neural circuits in cortical areas along the occipito-temporo-hippocampal pathway in the adult macaque monkey.

Animals↗

Distribution of [3H]kainic acid and binding sites in the rat brain: in vivo and in vitro receptor autoradiography.

Brain sections incubated in vitro with alpha-[3H]kainic acid (KA; spec. act. 62.5 Ci/mmol), reveal a heterogeneous distribution of low and high affinity KA binding sites in the brain. The highest density of KA binding sites was localised to the hippocampus CA3 region and to superficial layers of the entorhinal cortex (3.8-6.0 mu Ci/g tissue). Intravenous injection of [3H]KA (1 mu Ci/g) reveals limited overall penetration of [3H]KA across the blood-brain barrier. However, a dense labelling of the hippocampus, entorhinal cortex and lateral septal regions (2.5-3.8 mu Ci/g tissue) was observed. Behaviourally, these rats exhibited mild limbic seizure activity possibly as a result of a direct action of KA in the hippocampus or entorhinal cortex.

Animals↗

Regional properties of dorsal and ventral hippocampus in suppression of intralaminar thalamic unit responses.

In chloralose-anaesthetized, Flaxedil-paralysed cats, the suppression of extra-lemniscal thalamic units by dorsal and ventral hippocampus was investigated. Unitary responses to test somatic stimuli, recorded in centrolateral and neighbouring thalamic nuclei, were interacted with conditioning electric stimulation in different regions around the hippocampal arch, including parahippocampal gyrus (entorhinal and retrosplenial areas). Stimulation of dorsal (dhc) and ventral (VHC) hippocampus suppressed roughly equal proportions of responses. However, within each of DHC and VHC, effectiveness depended on the region stimulated. In DHC, fields CA1 and CA3, subiculum (SUB), and retrosplenial area, but not field CA4 or dentate gyrus, usually suppressed extralemniscal units at currents below 1.0 mA. In VHC, the most effective regions were entorhinal cortex, CA3, and CA4 with dentate gryus (FD), while stimulation of CA1 or subiculum was almost ineffective, at currents below 1.0 mA. In VHC, the regions were ranked for effectiveness: Entorhinal cortex=CA3 is greater than FD is greater than SUB is greater than CA1. No topographic relationship was found between hippocampal region and thalamic loci for unit suppression. Lemniscal-type unit responses in ventrobasal thalamus were unaffected by stimulation of the hippocampus or parahippocampal gryus. Interruption of the fornix-fimbria system prevented suppression elicited from CA1 of DHC, or from CA3 but not FD of VHC. It had no effect on suppression elicited from retrosplenial or entorhinal cortex. Hippocampal regional variation of effectiveness in suppressing extralemniscal pathways may contribute to the differential behavioural involvements reported for different hippocampal structures.

Animals↗

Effect of the selective lesion of serotoninergic neurons on the regional distribution of 5-HT1A receptor mRNA in the rat brain.

The effects of the selective lesion of serotoninergic neurons by an intra-raphe administration of 5,7-dihydroxytryptamine on the 5-HT1A receptor protein and the 5-HT1A receptor mRNA were examined in various regions of the rat brain using specific antibodies and an antisense riboprobe, respectively. Twenty one days after the treatment, the 5-HT1A receptor protein was no longer detected within the dorsal raphe nucleus but was still present in the hippocampus and entorhinal cortex. Quantitative in situ hybridization showed an 85% decrease in the levels of 5-HT1A receptor mRNA within the dorsal raphe nucleus, but no significant change in the hippocampus, interpeduncular nucleus and entorhinal cortex of 5,7-dihydroxytryptamine-treated rats. These data demonstrate that 5-HT1A receptors are synthesized by serotoninergic neurons in the dorsal raphe nucleus, and by neurons located postsynaptically with regard to serotoninergic projections in other areas. The unchanged levels of 5-HT1A receptor mRNA in the hippocampus, interpeduncular nucleus and entorhinal cortex three weeks after the extensive lesion of serotoninergic neurons are consistent with the absence of 5-HT1A receptor up regulation already reported under this condition.

5,7-Dihydroxytryptamine↗

Region specific neuron loss in the aged canine hippocampus is reduced by enrichment.

Neuron loss within the hippocampus and entorhinal cortex occurs as a function of age in humans. We first tested the hypothesis that neuron loss occurs in the aged dog. The total unilateral number of neurons in the canine entorhinal cortex and subdivisions of the hippocampus from the left hemisphere were estimated using the optical fractionator. The brains from 5 old (13.0-15.0 years old) and 5 young (3.4-4.5 years old) beagle dogs were analyzed. The hilus of the hippocampus showed a significant loss of neurons (approximately 30%) in the aged dog brain compared to young. Differences were not detected in the remaining hippocampal subfields and entorhinal cortex. We further tested the hypothesis that an antioxidant fortified food or behavioral enrichment would reduce the age-related loss of hilar neurons. Behaviorally enriched aged dogs had more neurons in the hilus (approximately 18%) compared to aged controls. These results suggest that the aged canine hippocampus in the left hemisphere shows selective neuron loss and that behavioral enrichment may reduce this loss.

Age Factors↗

Developmental differences in neural damage following trimethyl-tin as demonstrated with GFAP immunohistochemistry.

Long-Evans rat pups received intraperitoneal (i.p.) injections of trimethyl-tin (TMT) 6 mg/kg hydroxide or saline on postnatal day (PND) 10 or PND 18 and were sacrificed for immunohistochemical staining for glial fibrillary acidic protein (GFAP) on PND 12, 18, 20, or 25. After dosing with TMT on PND 10 there was a transient increase in GFAP immunoreactivity (IR) in the amygdala, piriform, and entorhinal cortex 2 days post-dosing (PND 12) and a persistent increase in GFAP IR in the hippocampus and cingulate cortex up to two weeks post-dosing. Following dosing with TMT on PND 18 there was a delayed (PND 25) increase in GFAP IR in the amygdala, hippocampus, cingulate, piriform, and entorhinal cortex. In addition, increases in GFAP IR were observed in the neocortex 7 days post-dosing, which was not observed following earlier postnatal dosing. The regions in which gliosis and loss of Nissl-staining were consistent for the different time points of TMT treatment were the amygdala, hippocampus, cingulate, piriform, and entorhinal cortex. The present findings indicate the GFAP immunohistochemistry can be used to reveal regional effects of developmental neurotoxicant exposure during early stages of development.

Animals↗

Dopamine D2 receptors in the rat, monkey and the post-mortem human hippocampus. An autoradiographic study using the novel D2-selective ligand 125I-NCQ 298.

The distribution of dopamine D2 receptors in the hippocampal region of the rat, monkey and the postmortem human brain was studied with in vitro receptor autoradiography using the selective salicylamide ligand 125I-NCQ 298. Specific binding was defined in the presence of the D2-selective compound raclopride. In all 3 species, higher densities of specifically bound 125I-NCQ 298 was found in the retrohippocampal structures than in the hippocampus proper. In the rat, layers 1 and 3 of the entorhinal cortex and layer 2 of the presubiculum were found to be rich in specific binding sites. In the monkey, the highest densities were detected in the deep layers (4 through 6) of the entorhinal cortex (EC) and in layer 2 of the presubiculum. Relatively high density of binding was found in the granule cell layer of area dentata. In the human brain, less specific binding was seen as compared to the other two species; the highest densities occurred in the outer layers of the presubiculum and in the hilus of area dentata. These findings show that D2 receptors are present in the hippocampal region and that the retrohippocampal region, including the entorhinal cortex, is enriched in dopamine D2 receptors.

Animals↗

Anomalous expression of microtubule-associated protein 1B in the hippocampus and cortex of aged rats treated with pentylenetetrazole.

The aim of the present study was to assess the age-dependent response of microtubule-associated protein 1B, a plasticity-associated protein deriving from a late gene, following administration of an epileptogenic stimulus. The effect of a single administration of the convulsant pentylenetetrazole on microtubule-associated protein 1B expression in the hippocampal formation and cortex of three-, 18- and 28-month-old rats was assessed using northern blot analysis, in situ hybridization and immunohistochemistry. In three-month-old rats, we detected initial increases in microtubule-associated protein 1B messenger RNA at 15 h following pentylenetetrazole administration in the granule cells of the dentate gyrus, in the CA3 region of the hippocampus and in layers II/III of the entorhinal cortex, and these reached a maximum at 44 h. However, in the hippocampus and cortex of 18-month-old rats, the peak occurred at 15 h, and in the brains of 28-month-old rats a blunted peak was reached at 3 h. Pentylenetetrazole treatment in young rats resulted in a robust induction of microtubule-associated protein 1B immunoreactivity in the granule cells of the dentate gyrus and in layers II/III of the entorhinal cortex, but also produced a large decrease in the retrosplenial cortex. However, following pentylenetetrazole treatment in older rats, the granule cells of the dentate gyrus were nearly devoid of microtubule-associated protein 1B immunoreactivity, whereas the retrosplenial cortex showed no changes at all, and the entorhinal cortex had an expression pattern similar to that of young rats. Aberrant immunolabeling of microtubule-associated protein 1B occurred in cortical layer VI of the aged rats where, unlike in young rats, there was heavy staining of neuronal somata. These results suggest that the regulation of the plasticity-associated protein microtubule-associated protein 1B is altered in the ageing rat brain, with the peak of expression shifted to earlier times in 18-month-old rats and blunted, variable increases at even earlier times in 28-month-old rats.

Aging↗