The effects of arousal on apical amplification and conscious state.

Neocortical pyramidal cells can integrate two classes of input separately and use one to modulate response to the other. Their tuft dendrites are electrotonically separated from basal dendrites and soma by the apical dendrite, and apical hyperpolarization-activated currents (Ih) further isolate subthreshold integration of tuft inputs. When apical depolarization exceeds a threshold, however, it can enhance response to the basal inputs that specify the cell's selective sensitivity. This process is referred to as apical amplification (AA). We review evidence suggesting that, by regulating Ih in the apical compartments, adrenergic arousal controls the coupling between apical and somatic integration zones thus modifying cognitive capabilities closely associated with consciousness. Evidence relating AA to schizophrenia, sleep, and anesthesia is reviewed, and we assess theories that emphasize the relevance of AA to consciousness. Implications for theories of neocortical computation that emphasize context-sensitive modulation are summarized. We conclude that the findings concerning AA and its regulation by arousal offer a new perspective on states of consciousness, the function and evolution of neocortex, and psychopathology. Many issues worthy of closer examination arise.

The neurobiology of infant maternal odor learning

Infant rats must learn to identify their mother’s diet-dependent odor. Once learned, maternal odor controls pups’ approach to the mother, their social behavior and nipple attachment. Here we present a review of the research from four different laboratories, which suggests that neural and behavioral responses to the natural maternal odor and neonatal learned odors are similar. Together, these data indicate that pups have a unique learning circuit relying on the olfactory bulb for neural plasticity and on the hyperfunctioning noradrenergic locus coeruleus flooding the olfactory bulb with norepinephrine to support the neural changes. Another important factor making this system unique is the inability of the amygdala to become incorporated into the infant learning circuit. Thus, infant rats appear to be primed in early life to learn odors that will evoke approach responses supporting attachment to the caregiver.

The neurobiology of infant maternal odor learning.

Infant rats must learn to identify their mother's diet-dependent odor. Once learned, maternal odor controls pups' approach to the mother, their social behavior and nipple attachment. Here we present a review of the research from four different laboratories, which suggests that neural and behavioral responses to the natural maternal odor and neonatal learned odors are similar. Together, these data indicate that pups have a unique learning circuit relying on the olfactory bulb for neural plasticity and on the hyperfunctioning noradrenergic locus coeruleus flooding the olfactory bulb with norepinephrine to support the neural changes. Another important factor making this system unique is the inability of the amygdala to become incorporated into the infant learning circuit. Thus, infant rats appear to be primed in early life to learn odors that will evoke approach responses supporting attachment to the caregiver.

Calcineurin inhibition eliminates the normal inverted U curve, enhances acquisition and prolongs memory in a mammalian 3'-5'-cyclic AMP-dependent learning paradigm.

The role protein phosphatase 2B (calcineurin, CaN) plays in learning and memory has received a significant amount of attention due to its promotion of the dephosphorylation of 3'-5'-cyclic AMP response element binding protein (CREB). Researchers have ascertained that overexpression of CaN is associated with memory retention deficits [Foster TC, Sharrow KM, Masse JR, Norris CM, Kumar A (2001) Calcineurin links Ca(2+) dysregulation with brain aging. J Neurosci 21:4066-4073; Mansuy IM, Mayford M, Jacob B, Kandel ER, Bach ME (1998) Restricted and regulated overexpression reveals calcineurin as a key component in the transition from short-term to long-term memory. Cell 92:39-49], while CaN inhibition enhances learning and memory [Gerdjikov TV, Beninger RJ (2005) Differential effects of calcineurin inhibition and protein kinase A activation on nucleus accumbens amphetamine-produced conditioned place preference in rats. Eur J Neurosci 22:697-705; Ikegami S, Inokuchi K (2000) Antisense DNA against calcineurin facilitates memory in contextual fear conditioning by lowering the threshold for hippocampal long-term potentiation induction. Neuroscience 98:637-646]. The present study hypothesized that infusion of a CaN inhibitor (FK506) bilaterally into the olfactory bulbs of postnatal day 6 Sprague Dawley rat pups would prolong the duration of a conditioned odor preference and retard cyclic AMP response element binding protein dephosphorylation. A 2 mg/kg s.c. injection of isoproterenol (ISO, beta-adrenoceptor agonist) was paired with a 10 min exposure to peppermint and subsequently an infusion of FK506. Immunohistochemistry for phosphorylated 3'-5'-cyclic AMP response element binding protein (pCREB) revealed that unilateral infusion of FK506 resulted in an amplification of phosphorylated CREB in the olfactory bulb 40 min after training compared with saline-infused bulbs. Pups infused bilaterally with FK506 maintained a learned preference for peppermint 48, 72 and 96 h after training. CaN inhibition also modified the conventional inverted U curve obtained when ISO is used to replace stroking, as the unconditioned stimulus. When pups were infused with FK506, learning occurred with sub- and supra-optimal doses of ISO indicating that CaN overcomes non-optimal effects ISO may have on learning. We demonstrate that CaN inhibition can extend the duration of conditioned olfactory memory and may provide a target for memory prolongation that is superior to even phosphodiesterase inhibition observed in previous studies.

Potentiation and prolongation of long-term odor memory in neonate rats using a phosphodiesterase inhibitor.

Cyclic AMP has been shown to have a critical role in learning and memory in invertebrates. Here we use the rat pup odor preference learning model in which odor acts as a conditioned stimulus and beta-adrenoceptor stimulation acts as an unconditioned stimulus to test the role of cyclic AMP in an associative mammalian paradigm. A phosphodiesterase inhibitor that prevents cyclic AMP breakdown (cilomilast) makes a low, learning-ineffective dose of a beta-adrenoceptor agonist (isoproterenol, 1mg/kg) an effective unconditioned stimulus in pup odor preference learning. A dose of the phosphodiesterase inhibitor (cilomilast, 1 mg/kg) that induces learning with a weak unconditioned stimulus interferes with learning using a normally optimal unconditioned stimulus (isoproterenol, 2 mg/kg). Cilomilast (3 mg/kg) paired with peppermint odor during learning, prolonged memory at least four times longer than without the drug (24 h vs. 96 h). These data demonstrate a causal role for cyclic AMP in the acquisition and duration of odor preference learning in the rat pup.

Infusion of the metabotropic receptor agonist, DCG-IV, into the main olfactory bulb induces olfactory preference learning in rat pups.

DCG-IV, a type 2 metabotropic glutamate receptor (mGluR2) agonist, was infused into the main olfactory bulb of 1-week-old pups exposed to peppermint odor. A preference for peppermint was demonstrated 24 h later. The data support the proposal that disinhibition at dendrodendritic synapses between granule cells and mitral cells is a critical component of olfactory memory formation in the rat pup olfactory bulb as well as in the accessory olfactory bulb of adult rodents.

Daily variation in the distribution of glycogen phosphorylase in the suprachiasmatic nucleus of Syrian hamsters.

Dynamic changes in astrocytic processes in the Syrian hamster suprachiasmatic nucleus (SCN) have been reported with maximal process extension in the light phase and maximal process retraction in the dark phase of a daily light:dark cycle. In the present study, we asked whether dynamic changes occur in the distribution of an astrocytic metabolic marker, glycogen phosphorylase (GP), using a histochemical assay to reveal the distribution of both active and total GP, in the hamster SCN. Changes in glial acidic fibrillary protein (GFAP) immunoreactivity also were assessed using a relative optical density measure (ROD). We observed changes in the localization and distribution of GP both in the SCN and in the paraventricular nucleus of the hypothalamus (PVN) as a function of time of day. In the light phase, there were concentrated, large, dot-like deposits of GP throughout the SCN and PVN on an empty background. In the dark phase, diffuse, small, granular particles were seen throughout both nuclei. Selectively, in the dark-phase SCN, these granular particles formed a rim of intense GP reactivity on the lateral, ventral, posterior, and medial borders. Significantly higher levels of GP reactivity were seen in anterior sections of the medial optic chiasm in the light phase. GFAP-immunoreactive astrocytic processes had higher ROD levels in the dark phase. In conclusion, the astrocytic metabolic marker, GP, exhibits a significant daily variation in localization in both the SCN and the PVN that correlates with dynamic changes in the distribution of astrocytic processes in the SCN. Increased GP activity also occurs in astrocytes among optic fibers subjacent to the SCN during light input.

Spatial problem solving and hippocampal place cell firing in rats: control by an internal sense of direction carried across environments.

Rats learned to find the baited corner of a box surrounded by a curtain, regardless of whether they had a fixed or random point of entry (POE) through the curtain. On probe trials, rats used an internal direction sense carried from outside the curtain to solve the problem, and only used the visual cue inside the curtain if disoriented and denied access to a view of the room en route. Similar disorientation procedures were required to obtain cue control of hippocampal place fields. The results suggest that: (1) POE effects previously found in the water maze may be task-specific; (2) an undisrupted internal sense of direction carried from one environment to another may provide the preferred solution to spatial problems in the second environment, even when this second environment is a familiar one with stable visual cues; and (3) choice behaviour is sometimes, but not always, representative of the hippocampal representation of space.

The moving fire hydrant experiment: movement of objects to a new location reelicits marking in rats.

Three experiments were carried out to investigate the influence of object relocation on object marking in an open field by hooded and albino rats. Object marking was reelicited when an object was moved to a new location in the second half of an open field test. Control conditions revealed that an object briefly moved and returned to the original location elicited no more marking than a stationary object. The higher level of marking of the relocated object suggests that object marking may provide an index of spatial knowledge. The implication of spatial knowledge in controlling marking behavior is congruent with observations that rats with hippocampal damage show increased marking.

Glycogen phosphorylase reactivity in the amygdala and bed nucleus of the stria terminalis.

The present study examines the reactivity of the glial metabolic enzyme, glycogen phosphorylase, within the amygdala and bed nucleus of the stria terminalis. Reactivity for phosphorylase a, the active form of glycogen phosphorylase, was higher in all parts of the medial amygdaloid nucleus, in the medial division of the central amygdaloid nucleus, in the anterior amygdaloid area and in the bed nucleus of stria terminalis than in all parts of the lateral amygdaloid nucleus, the anterior cortical amygdaloid nucleus, the posteromedial and posterolateral cortical amygdaloid nuclei, the intercalated nucleus of the amygdala, main part and the intercalated nuclei. A greater degree of phosphorylase a reactivity was also observed in the basolateral amygdaloid nucleus, anterior and posterior parts, and in the basomedial amygdaloid nucleus, anterior part, while other parts of these nuclei were less reactive. Reactivity attributed to total glycogen phosphorylase enzyme, phosphorylase a+phosphorylase b activated by AMP, was higher and homogeneous across the amygdala. Phosphorylase a patterns are likely to reflect differences in the contribution of glycogenolysis to the metabolic support of cells in the amygdala and bed nucleus of the stria terminalis. Possible relationships to local neuronal activity and to differences in glycogenolytic neuromodulatory input are discussed.

A two-platform task reveals a deficit in the ability of rats to return to the start location in the water maze.

The ability of rats to return to the start location was examined with a 4-arm radial water maze. The task required rats to find 2 hidden platforms in sequence. Rats were released from 1 of 3 arms and there was a platform located in the fourth arm. Once a rat found this platform, a 2nd platform was raised in another location, which was either the start location, for 1 group, or another fixed location, for a control group. Across 3 experiments, all rats learned the location of the 1st fixed platform in 80 to 120 trials. However, rats had difficulty finding a 2nd platform if it was at the start location. Control groups revealed that rats could learn 2 platform locations and that the difficulty in learning to return to the start location did not seem to be attributable to its aversive nature. In separate groups, exposure to the start location was increased by starting the rats from an initially stable platform. Rats still did not readily learn to return to the start location. The authors suggest that start location, when varied, cannot readily be used to define the location of a hidden platform.

Beta-adrenergic blockade in the dentate gyrus in vivo prevents high frequency-induced long-term potentiation of EPSP slope, but not long-term potentiation of population spike amplitude.

High frequency (HF)-induced and norepinephrine (NE)-induced long-term potentiation have been hypothesized to utilize common mechanisms of induction and expression in the dentate gyrus. In vitro data tend to support this hypothesis, but few studies have been done in vivo. The present study records perforant path-evoked potentials simultaneously on two micropipettes, one filled with saline and the other with the beta-antagonist, timolol. Stimulation of the paragigantocellularis nucleus (PGi) was used as a method of producing NE release in the dentate gyrus, and thus, to assess the efficacy of beta-receptor blockade on the timolol pipette. Beta-blockade by timolol attenuated PGi-induced spike potentiation. HF-induced potentiation of the excitatory post-synaptic potential (EPSP) slope was also blocked by timolol, but HF-induced spike amplitude potentiation was unaffected. These results are consistent with an earlier report examining HF-long-term potentiation (LTP) following 6-OHDA-induced NE depletion, which showed that the EPSP slope LTP depended, for its full expression, on NE, but potentiation of the population spike amplitude component of HF-induced LTP did not. In the present study, PGi-induced potentiation of spike amplitude on the saline pipette was normal after HF-induced saturation of spike amplitude potentiation, suggesting that the mechanisms for expression of spike potentiation, as well as induction of spike potentiation, are separate for HF and NE stimulation.

Isoproterenol increases CREB phosphorylation and olfactory nerve-evoked potentials in normal and 5-HT-depleted olfactory bulbs in rat pups only at doses that produce odor preference learning.

Norepinephrine (NE) and serotonin (5-HT) are important modulators of early odor preference learning. NE can act as an unconditioned stimulus (UCS), whereas 5-HT facilitates noradrenergic actions. In this study, we examined the phosphorylation of an important transcription factor, cAMP response element binding protein (CREB), which has been implicated in long-term-memory formation (McLean et al. 1999) during NE-induced odor preference learning in normal and olfactory bulb 5-HT-depleted rat pups. We also examined NE modulation of olfactory nerve-evoked field potentials (ON-EFPs) in anesthetized normal and bulbar 5-HT depleted pups. Systemic injection of 2 mg/kg isoproterenol (beta-adrenoceptor agonist) induced odor preference learning, enhanced pCREB expression in the olfactory bulbs at 10 min after odor pairing, and increased ON-EFPs in normal rat pups but not in bulbar 5-HT-depleted rat pups. A dose of 6 mg/kg isoproterenol, which was ineffective in modulating these measures in normal rat pups, induced odor preference learning, enhanced phosphorylated CREB (pCREB) expression, and increased ON-EFPs in bulbar 5-HT-depleted pups. These outcomes suggest that NE and 5-HT promote specific biochemical and electrophysiological changes that may critically underlie odor preference learning.

Glutamatergic activation of the medial septum complex: an enhancement of the dentate gyrus population spike and accompanying EEG and unit changes.

A large number of cells from the medial septum complex (MSC) innervate the dentate gyrus of the hippocampus. Electrical prestimulation of the MSC enhances perforant path-dentate gyrus evoked field potentials. Considering the large number of fibres that pass through this region, the effects glutamatergic stimulation of the MSC had on dentate gyrus field potentials, and accompanying changes in units, and EEG, was investigated in urethane-anaesthetized rats. The perforant path was stimulated at a rate of 0.1 Hz, evoking an EPSP and a population spike recorded in the dentate gyrus granule cell layer. L-glutamate was delivered by pressure ejection. Glutamate ejection to the MSC produced a significant enhancement of the population spike. The duration of enhancement ranged from 1 to 49 min ( approximately =10.5 min). A consistent, but relatively short increase in the EPSP slope was also demonstrated. MSC activation induced a theta rhythm in 7 of 10 animals (duration=20-112 s). Theta rhythm induction preceded spike enhancement and occurred for a shorter duration than the enhancement. The effects on spontaneous unit activity were mixed. However, all changes in firing rate preceded spike enhancement, and their duration rarely coincided with the duration of the spike enhancement. The population spike enhancement usually occurred without evidence of a change in paired-pulse inhibition.

Multiple points of entry into a circular enclosure prevent place learning despite normal vestibular orientation and cue arrays: evidence for map resetting.

This study identified sources of map orientation critical for successful spatial problem solving by rats of a plus maze embedded in water. Disorientation slowed, but it did not prevent acquisition of goal location. Use of a circular enclosure with multiple points of entry prevented reliable goal location. A single entry point enabled the rats to locate a fixed goal. A cue array within the enclosure was ineffective in providing orientation. These data suggest that stable map orientation can be derived from entry location when enclosure geometry is uniformative, but is not readily taken from cue arrays. They further suggest that map orientation is reset when rats enter an enclosure.

Testosterone decreases CA1 plasticity in vivo in gonadectomized male rats.

Estrogen has been reported to enhance CA1 functional plasticity in adult rats, as measured by the induction of long-term potentiation (LTP). In the present study, the effects of androgens on CA1 LTP were assessed in adult male Sprague-Dawley rats in vivo following peripubertal castration. Castrated rats with cholesterol implants showed significantly greater plasticity, both in degree and duration of potentiation, than castrated rats given testosterone or dihydrotestosterone implants. An LTP paradigm reported to produce decremental LTP in vitro produced nondecremental LTP in androgen-deprived rats, but decremental LTP in androgen-treated rats. These results suggest that androgens, unlike estrogens, act to reduce CA1 plasticity in the adult rat.

Open field motor patterns and object marking, but not object sniffing, are altered by ibotenate lesions of the hippocampus.

Objectmarking and object sniffing were assessed in an open field during six 5-min trials in unoperated Long-Evans rats and rats with ibotenate lesions of the hippocampus and/or neocortex. Object marking was higher in hippocampally lesioned rats than in unoperated rats. Object marking did not differ between neocortically lesioned rats and unoperated rats. Object sniffing durations and visits did not differ between unoperated and hippocampally lesioned rats nor between unoperated and neocortically lesioned rats. A new object elicited longer sniffing by both unoperated and hippocampally lesioned rats. Neocortically lesioned rats did not show this effect. There were no effects of the new object on marking. Computerized tracing of open field paths revealed a smaller perimeter track for hippocampally lesioned rats than for unoperated rats. This difference reflected distinct ambulatory patterns. Hippocampally lesioned rats made stereotyped hind-limb pivots at each corner, while normal rats used forelimb pivots or reared and reoriented adjacent to the wall. Rearing was lower in hippocampally lesioned rats, and higher in neocortically lesioned rats, than in unoperated rats. These data indicate that investigative object behavior (sniffing) is resistant to the effects of hippocampal damage, while object-elicited marking and motoric output may be profoundly altered. The data on sniffing suggest either that (1) the noticeability of the objects used elicited investigative behaviors in hippocampally damaged rats comparable to those of novelty-induced exploration in normal rats or (2) object exploration is not used to create a spatial map, and, hence, not disturbed by hippocampal loss. Object marking may require spatial locale information to be exercised normally, or may index the mediation of an olfactory-modulated behavioral pattern through the hippocampal system.

Hippocampal volume is related to complexity of nesting habitat in Leach's storm-petrel, a nocturnal procellariiform seabird.

The hippocampal and telencephalon volumes of the nocturnal Leach's storm-petrel (Oceanodroma leucorhoa, n = 15) were compared with published data for food-storing and non-storing Passerines. The hippocampus to telencephalon ratio of Leach's storm-petrels is intermediate between food-storing and non-storing birds. Leach's storm-petrels taken from nesting burrows in wooded habitat had a larger relative hippocampal volume than those taken from burrows in an open meadow. Relative olfactory volume did not differ between woods and open-nesting storm-petrels. The larger relative hippocampal volume of storm-petrels may be associated with increased spatial demands of returning to their nests at night in the darker, more navigationally complex woods. It is not known whether the larger hippocampus in storm-petrels from the woods is due to selection on different subpopulations or whether experience in a more complex environment results in greater hippocampal volume. Hippocampal volume from the brain of one diurnal Procellariiforme, the northern fulmar (Fulmaris glacialis), fell within the range of non-storing species, which supports the view that hippocampal enlargement in the storm-petrel is related to the spatial demand of returning to the nest at night.

pCREB in the neonate rat olfactory bulb is selectively and transiently increased by odor preference-conditioned training.

Early olfactory preference learning in rat pups occurs when novel odors are paired with tactile stimulation, for example stroking. cAMP-triggered phosphorylation of cAMP response element binding protein (pCREB) has been implicated as a mediator of learning and memory changes in various animals (Frank and Greenberg 1994). In the present study we investigate whether CREB is phosphorylated in response to conditioned olfactory training as might be predicted given the proposed role of the phosphorylated protein in learning. On postnatal day 6, pups were trained for 10 min using a standard conditioned olfactory learning paradigm in which a conditioned stimulus, Odor, was either used alone or paired with an unconditioned stimulus, Stroking (using a fine brush to stroke the pup). In some instances stroking only was used. The pups were sacrificed at 0, 10, 30, or 60 min after the training. Using Western blot analysis, we observed that the majority of olfactory bulbs in conditioned pups (Odor + Stroking) had a greater increase in pCREB activation at 10 min after training than pups given nonlearning training (Odor only or Stroking only). The phosphorylated protein levels were low at 0 min and at 60 min after training. This is in keeping with the slightly delayed and short-lived activation period for this protein. The localization of pCREB increases within the olfactory bulb as seen by immunocytochemistry. Naive pups were not exposed to odor or training. There was a significantly higher level of label in mitral cell nuclei within the dorsolateral quadrant of the bulb of pups undergoing odor-stroke pairing. No significant differences were observed among nonlearning groups (Naive, Odor only, or Stroking only) or among any training groups in the granule or periglomerular cells of the dorsolateral region. The localized changes in the nuclear protein are consistent with studies showing localized changes in the bulb in response to a learned familiar odor. The present study demonstrates that selective increases in pCREB occur as an early step following pairing procedures that normally lead to the development of long-term olfactory memories in rat pups. These results support the hypothesized link between pCREB and memory formation.