Heather Dickinson-Anson

Neurogenesis in the dentate gyrus of the adult rat: age-related decrease of neuronal progenitor proliferation

The hippocampus is one of the few areas of the rodent brain that continues to produce neurons postnatally. Neurogenesis reportedly persists in rats up to 11 months of age. Using bromodeoxyuridine (BrdU) labeling, the present study confirms that in the adult rat brain, neuronal progenitor cells divide at the border between the hilus and the granule cell layer (GCL). In adult rats, the progeny of these cells migrate into the GCL and express the neuronal markers NeuN and calbindin-D28k. However, neurogenesis was drastically reduced in aged rats. Six-to 27-month-old Fischer rats were injected intraperitoneally with BrdU to detect newborn cells in vivo and to follow their fate in the dentate gyrus. When killed 4–6 weeks after BrdU labeling, 12- to 27- month-old rats exhibited a significant decline in the density of BrdU- positive cells in the granule cell layer compared with 6-month-old controls. Decreased neurogenesis in aging rats was accompanied by reduced immunoreactivity for poly-sialylated neural cell adhesion molecule, a molecule that is involved in migration and process elongation of developing neurons. When animals were killed immediately (12 hr) after BrdU injection, significantly fewer labeled cells were observed in the GCL and adjacent subgranular zone of aged rats, indicative of a decrease in mitotic activity of neuronal precursor cells. The reduced proliferation was not attributable to a general aged- related metabolic impairment, because the density of BrdU-positive cells was not altered in other brain regions with known mitotic activity (e.g., hilus and lateral ventricle wall). The decline in neurogenesis that occurs throughout the lifespan of an animal can thus be related to a decreasing proliferation of granule cell precursors.

Altered synaptic physiology and reduced susceptibility to kainate-induced seizures in GluR6-deficient mice

l-glutamate, the neurotransmitter of the majority of excitatory synapses in the brain, acts on three classes of ionotropic receptors: NMDA (N-methyl-d-aspartate), AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid) and kainate receptors. Little is known about the physiological role of kainate receptors because in many experimental situations it is not possible to distinguish them from AMPA receptors,. Mice with disrupted kainate receptor genes enable the study of the specific role of kainate receptors in synaptic transmission as well as in the neurotoxic effects of kainate. We have now generated mutant mice lacking the kainate-receptor subunit GluR6. The hippocampal neurons in the CA3 region of these mutant mice are much less sensitive to kainate. In addition, a postsynaptic kainate current evoked in CA3 neurons by a train of stimulation of the mossy fibre system is absent in the mutant,. We find that GluR6-deficient mice are less susceptible to systemic administration of kainate, as judged by onset of seizures and by the activation of immediate early genes in the hippocampus. Our results indicate that kainate receptors containing the GluR6 subunit are important in synaptic transmission as well as in the epileptogenic effects of kainate.

Behavioral and neuroanatomical investigation of Highly Superior Autobiographical Memory (HSAM)

A single case study recently documented one womans ability to recall accurately vast amounts of autobiographical information, spanning most of her lifetime, without the use of practiced mnemonics (Parker, Cahill, & McGaugh, 2006). The current study reports findings based on eleven participants expressing this same memory ability, now referred to as Highly Superior Autobiographical Memory (HSAM). Participants were identified and subsequently characterized based on screening for memory of public events. They were then tested for personal autobiographical memories as well as for memory assessed by laboratory memory tests. Additionally, whole-brain structural MRI scans were obtained. Results indicated that HSAM participants performed significantly better at recalling public as well as personal autobiographical events as well as the days and dates on which these events occurred. However, their performance was comparable to age- and sex-matched controls on most standard laboratory memory tests. Neuroanatomical results identified nine structures as being morphologically different from those of control participants. The study of HSAM may provide new insights into the neurobiology of autobiographical memory.

Hippocampal grafts of acetylcholine-producing cells are sufficient to improve behavioural performance following a unilateral Fimbria-Fornix lesion

Lesions of the septohippocampal pathway produce cognitive deficits that are partially attenuated by grafts of cholinergic-rich tissue into denervated target regions or by systemic administration of cholinomimetic drugs. In the present study, fibroblasts engineered to produce acetylcholine were used to test the hypothesis that restoration of hippocampal acetylcholine in rats with septohippocampal lesions is sufficient to improve cognitive processing post-damage. Rats received unilateral grafts of acetylcholine-producing or control fibroblasts into the hippocampus immediately prior to an aspirative lesion of the ipsilateral fimbria-fornix. Some rats with fimbria-fornix lesions were implanted with acetylcholine-producing or control fibroblasts into the neocortex, another major target of the basal forebrain cholinergic system, to determine if the site of acetylcholine delivery to the damaged brain is critical for functional recovery. Rats were tested in a hidden platform water maze task, a cued water maze task and activity chambers between one and three weeks post-grafting. Compared to unoperated controls, rats with fimbria fornix lesions only were significantly impaired in hidden platform water maze performance. Hippocampal grafts of acetylcholine-producing cells reduced lesion-induced deficits in the water maze, whereas hippocampal control grafts and cortical grafts of either cell type were without effect. Locomotor activity and cued water maze performance were unaffected by the lesion or the implants. Taken together, these data indicate that water maze deficits produced by fimbria fornix lesions, which disrupt a number of hippocampal neurotransmitter systems, can be attenuated by target specific replacement of acetylcholine in the hippocampus and that this recovery occurs in the absence of circuitry repair.

Bicuculline administered into the amygdala after training blocks benzodiazepine-induced amnesia

Male Sprague-Dawley rats were injected (i.p.) with either midazolam (MDZ, 2.0 mg/kg) or vehicle (1.0 ml/kg) 10 min before they were trained on a multiple-trial inhibitory avoidance task. Immediately following the training, bicuculline methiodide (BMI; 2.0, 5.6, 56.0 or 197.0 pmol/0.5 microl) or vehicle (0.5 microl) was infused bilaterally into the amygdala. On a 48 h retention test the performance of the MDZ-treated animals was significantly poorer than that of controls. The retention of MDZ-treated animals given intra-amygdala injections of the lowest dose of BMI (2.0 pmol) was comparable to that of controls, whereas higher doses of BMI impaired retention. The present results are consistent with other findings indicating that the amygdala mediates the amnestic effects of benzodiazepines on aversive learning. Furthermore, these data suggest that benzodiazepines impair memory by disrupting post-training processes underlying memory consolidation.

Amygdala lesions block the amnestic effects of diazepam

This experiment examined the effects of pre-training systemic injections of the benzodiazepine (BZ) diazepam (DZP) on learning and retention of an inhibitory avoidance response in rats with bilateral lesions of the amygdaloid complex (AC) induced by intra-amygdala injections of the excitotoxin N-methyl-D-aspartic acid (NMDA). Unoperated, sham-operated and AC-lesioned rats received i.p. injections of DZP (1.0 or 2.0 mg/kg) or vehicle 30 min prior to training in a continuous multiple-trial inhibitory avoidance task. Retention was tested 48 h later. The acquisition and retention of the AC-lesioned rats were impaired, relative to that of the unoperated and sham controls. In the unoperated and sham controls, DZP impaired retention but did not affect acquisition. In contrast, in animals with AC lesions, DZP did not affect either acquisition or retention. These findings suggest that the amnestic effects of DZP are mediated, at least in part, through influences involving the AC.

Lesions of the basolateral amygdala complex block propofol-induced amnesia for inhibitory avoidance learning in rats.

BackgroundAs the unitary theory of anesthesia gives way to the “multiple sites, multiple mechanisms” concept, the sites involved in mediating the components of anesthesia must be identified. In the current study, we test the hypothesis that the basolateral amygdala complex (BLAC) is a brain site involved with mediating propofol-induced amnesia. MethodsMale Sprague-Dawley rats were divided into two groups, sham-operated control animals and rats given bilateral excitotoxic N-methyl-d-aspartate lesions of the BLAC. For each group, animals were given intraperitoneal saline or propofol (25 mg/kg) 5 min before inhibitory avoidance learning. Rats were given a foot shock (0.4 mA) upon entering the dark side of a two-sided apparatus. Rats could escape additional shock by returning to and staying in the light side. Training ended after shock avoidance for greater than 60 s. Memory was tested at 24 h. Longer latencies to enter the dark side 24 h after training imply better memory. ResultsSham-saline–treated animals had a robust memory latency (median latency [interquartile range] = 300 [163–567] s). Sham-propofol–treated animals exhibited a significant anterograde amnesia (latency = 63 [14–111] s) (P < 0.05 vs. sham-saline–treated animal). Both the saline-injected and propofol-injected animals with BLAC lesions showed robust memory (latency = 300 [264–485] and 323 [143–480] s, respectively). These latencies did not differ from performance in the sham-saline–treated group and were significantly higher than the latency of the sham-propofol–treated group (both P < 0.05). ConclusionsDiscrete BLAC lesions blocked the amnestic effect of propofol. BLAC activity appears to be a requirement for propofol-induced amnesia. This finding suggests that the BLAC is a key brain site mediating anesthetic-induced amnesia.

Bicuculline administered into the amygdala blocks benzodiazepine-induced amnesia

This experiment investigated the effect of intra-amygdala administration of the GABAergic antagonist bicuculline methiodide on benzodiazepine-induced amnesia. Male Sprague-Dawley rats were implanted bilaterally with cannulae aimed at the amygdala and allowed to recover for 1 week. Ten minutes before training in a continuous multiple trial inhibitory avoidance task a buffer solution or bicuculline methiodide (56 pmol/0.5 microliters) was injected bilaterally into the amygdala and this injection was immediately followed by a systemic injection of saline or midazolam (1.0 mg/kg). In comparison with saline controls, midazolam-treated animals required more trials to reach the acquisition criterion of remaining in the starting chamber for 100 s. The midazolam effect on acquisition was not attenuated by intra-amygdala infusion of bicuculline methiodide, suggesting that the midazolam-induced changes in acquisition behavior do not involve the amygdaloid GABAergic system. On a 48-h retention test the performance of the midazolam-treated animals was significantly poorer than that of the controls. However, the retention performance of animals given intra-amygdala injections of bicuculline methiodide prior to the systemic injection of midazolam was comparable to that of the saline controls. These results suggest that the amygdaloid GABAergic system mediates the impairing effects of midazolam on retention of inhibitory avoidance training.

Midazolam administered into the amygdala impairs retention of an inhibitory avoidance task

This experiment investigated the amnestic effects of injections of the benzodiazepine midazolam administered into the amygdala prior to training on an inhibitory avoidance task. Male Sprague-Dawley rats were implanted bilaterally with cannulae aimed at the amygdala. After 1 week recovery a buffer solution or midazolam (3 or 10 micrograms/0.5 microliters) was injected bilaterally 5 min before a single training trail in a two-compartment inhibitory avoidance apparatus. The pretraining intra-amygdala injections of midazolam did not affect the training step-through latencies. However, on a 48-h retention test the step-through latencies of the midazolam-treated animals were significantly lower than those of the buffer controls. These findings are consistent with other recent evidence indicating that the amygdala is involved in mediating the amnestic effects of benzodiazepines.

Acetylcholine-Secreting Cells Improve Age-Induced Memory Deficits

The present study examined whether aged rats with naturally occurring cognitive deficits in spatial learning and memory would benefit from local chronic supplementation of acetylcholine. Aged impaired and aged unimpaired rats were pretested in the water maze to characterize the extent of age-induced cognitive impairment. Groups were matched for extent of deficits. The animals subsequently received implants of either acetylcholine-releasing cells or control cells into the cortical and hippocampal target regions of the basal forebrain. One week postgrafting, spatial learning and memory were retested using the same behavioral procedure. All aged groups acquired the platform position more slowly than young controls. However, aged impaired rats grafted with acetylcholine-releasing cells performed significantly better than aged impaired rats with control grafts, and they did not differ from aged unimpaired groups. A spatial memory probe test revealed that memory for the escape platform location of the acetylcholine-grafted rats was significantly better than that of rats with control grafts and matched the performance of young controls. In vitro, biochemical and electrophysiological analyses of the engineered cells confirmed choline acetyltransferase activity and showed quantal release of acetylcholine from the transduced cells. In vivo, RT-PCR of microdissected grafts indicated that the engineered cells expressed the choline acetyltransferase transgene for up to 40 days postgrafting. These results indicate that locally restricted supplementation of acetylcholine into the two major target regions of the cholinergic basal forebrain of aged impaired rats ameliorates some age-related cognitive deficits.