Reduced K+ Channel Inactivation, Spike Broadening, and After-Hyperpolarization in Kvβ1.1-Deficient Mice with Impaired Learning

A-type K⁺ channels are known to regulate neuronal firing, but their role in repetitive firing and learning in mammals is not well characterized. To determine the contribution of the auxiliary K⁺ channel subunit Kvβ1.1 to A-type K⁺ currents and to study the physiological role of A-type K⁺ channels in repetitive firing and learning, we deleted the Kvβ1.1 gene in mice. The loss of Kvβ1.1 resulted in a reduced K⁺ current inactivation in hippocampal CA1 pyramidal neurons. Furthermore, in the mutant neurons, frequency-dependent spike broadening and the slow afterhyperpolarization (sAHP) were reduced. This suggests that Kvβ1.1-dependent A-type K⁺ channels contribute to frequency-dependent spike broadening and may regulate the sAHP by controlling Ca²⁺ influx during action potentials. The Kvβ1.1-deficient mice showed normal synaptic plasticity but were impaired in the learning of a water maze test and in the social transmission of food preference task, indicating that the Kvβ1.1 subunit contributes to certain types of learning and memory.     

Differential Induction of Long-Term Synaptic Facilitation by Spaced and Massed Applications of Serotonin at Sensory Neuron Synapses of Aplysia californica

Serotonin (5HT)-induced facilitation of synaptic transmission from tail sensory neurons (SNs) to motor neurons (MNs) in the marine mollusc Aplysia provides a cellular model of short- and long-term memory for behavioral sensitization of the tail withdrawal reflex. Synaptic facilitation at these synapses occurs in three temporal phases: short-term (STF, lasting minutes), intermediate-term (ITF, lasting more than an hour), and long-term (LTF, lasting >24 hr). STF, ITF, and LTF differ in their induction requirements: A single brief exposure of 5HT induces STF, whereas five applications are required for ITF and LTF. Moreover, STF and LTF can be induced independently.     

Overexpression of and RNA interference with the CCAAT enhancer-binding protein on long-term facilitation of Aplysia sensory to motor synapses

In the marine mollusk Aplysia, the CCAAT/enhancer-binding protein, ApC/EBP, serves as an immediate early gene in the consolidation of long-term facilitation in the synaptic connection between the sensory and motor neurons of the gill-withdrawal reflex. To further examine the role of ApC/EBP as a molecular switch of a stable form of long-term memory, we cloned the full-length coding regions of two alternatively spliced forms, the short and long form of ApC/EBP. Overexpression of each isoform by DNA microinjection resulted in a l6-fold increase in the expression of the coinjected luciferase reporter gene driven by an ERE promoter. In addition, when we overexpressed ApC/EBP in Aplysia sensory neurons, we found that the application of a single pulse of 5-HT that normally induced only short-term facilitation now induced long-term facilitation. Conversely, when we attempted to block the synthesis of native ApC/EBP by microinjecting double-strand RNA or antisense RNA, we blocked long-term facilitation in a sequence-specific manner. These data support the idea that ApC/EBP is both necessary and sufficient to consolidate short-term memory into long-term memory. Furthermore, our results suggest that this double-strand RNA interference provides a powerful tool in the study of the genes functioning in learning and memory in Aplysia by specifically inhibiting both the constitutive and induced expression of the genes.     

CA1 Long-Term Potentiation Is Diminished but Present in Hippocampal Slices from α-CaMKII Mutant Mice

Previous work has shown that mice missing the α-isoform of calcium–calmodulin-dependent protein kinase II (α-CaMKII) have a deficiency in CA1 hippocampal long-term potentiation (LTP). Follow-up studies on subsequent generations of these mutant mice in a novel inbred background by our laboratories have shown that whereas a deficiency in CA1 LTP is still present in α-CaMKII mutant mice, it is different both quantitatively and qualitatively from the deficiency first described. Mice of a mixed 129SvOla/SvJ;BALB/c;C57Bl/6 background derived from brother/sister mating of the α-CaMKII mutant line through multiple generations (>10) were produced by use of in vitro fertilization. Although LTP at 60 min post-tetanus was clearly deficient in these (−/−) α-CaMKII mice (42.6%, n=33) compared with (+/+) α-CaMKII control animals (81.7%, n=17), α-CaMKII mutant mice did show a significant level of LTP. The amount of LTP observed in α-CaMKII mutants was normally distributed, blocked by APV (2.7%, n=8), and did not correlate with age. Although this supports a role for α-CaMKII in CA1 LTP, it also suggests that a form of α-CaMKII-independent LTP is present in mice that could be dependent on another kinase, such as the β-isoform of CaMKII. A significant difference in input/output curves was also observed between (−/−) α-CaMKII and (+/+) α-CaMKII animals, suggesting that differences in synaptic transmission may be contributing to the LTP deficit in mutant mice. However, tetani of increasing frequency (50, 100, and 200 Hz) did not reveal a higher threshold for potentiation in (−/−) α-CaMKII mice compared with (+/+) α-CaMKII controls.     

Possible novel features of synaptic regulation during long-term facilitation in Aplysia

Most studies of molecular mechanisms of synaptic plasticity have focused on the sequence of changes either at individual synapses or in the cell nucleus. However, studies of long-term facilitation at Aplysia sensory neuron–motor neuron synapses in isolated cell culture suggest two additional features of facilitation. First, that there is also regulation of the number of synaptic contacts between two neurons, which may occur at the level of cell pair-specific branch points in the neuronal arbor. Branch points contain many molecules that are involved in protein synthesis-dependent long-term facilitation including neurotrophins and the RNA binding protein CPEB. Second, the regulation involves homeostatic feedback and tends to keep the total number of contacts between two neurons at a fairly constant level both at rest and following facilitation. That raises the question of how facilitation and homeostasis can coexist. A possible answer is suggested by the findings that they both involve spontaneous transmission and postsynaptic Ca²⁺, which can have bidirectional effects similar to LTP and LTD in hippocampus. In addition, long-term facilitation can involve a change in the set point of homeostasis, which could be encoded by plasticity molecules such as CPEB and/or PKM. A computational model based on these ideas can qualitatively simulate the basic features of both facilitation and homeostasis of the number of contacts.

Synaptic plasticity is a change in strength of the synaptic connection (postsynaptic potential or PSP) between neurons and includes increases during facilitation and decreases during depression. Plasticity is thought to underlie circuit formation during development and learning and memory in adults, and correspondingly to be defective in neurodevelopmental disorders including autism, ADHD, and schizophrenia as well as learning and memory disorders including Alzheimer''s, age-related memory loss, and drug addiction (Hawkins 2013; Hawkins et al. 2017). Most studies of molecular mechanisms of synaptic plasticity have focused on either changes at individual synapses or gene regulation in the cell nucleus. However, studies of long-term facilitation at Aplysia sensory neuron–motor neuron (SN–MN) synapses in isolated cell culture (Glanzman et al. 1990), sensitization in the intact animal (Wainwright et al. 2004), and long-term potentiation in hippocampal neurons (Antonova et al. 2001, 2009) have shown that there are also changes in the number of contacts between presynaptic varicosities and the postsynaptic neuron. We refer to these as synaptic contacts although not all of them are functional synapses (Kim et al. 2003). The number of contacts is thought to be an important determinant of the strength of the PSP (Zhang et al. 2003) and to be different for different neuron pairs. It also increases during long-term facilitation of the PSP and is thought to be a major determinant of the time course of the facilitation (Bailey and Chen 1989).As in other systems (Antonova et al. 2001, 2009; Holtmaat and Svoboda 2009), the contacts are dynamic and are continually being formed and eliminated, but the total number and the PSP remain fairly constant both at rest and during long-term facilitation (Miniaci et al. 2008; Chen et al. 2014). Furthermore, the number of contacts and the PSP return to baseline when maintenance of the facilitation is blocked, but the individual contacts are not all the same as they were before facilitation. These results have led some to suggest that memories are not stored at individual synaptic contacts, as is often supposed, but rather are stored in the nucleus (Chen et al. 2014). However, most of the previous experiments have involved a single SN and a single MN, so it has not been possible to examine the synapse specificity of the effects. Experiments with one SN and two MNs (Martin et al. 1997) or two SNs and 1 MN (Schacher et al. 1997) have shown that facilitation of the number of synaptic contacts and the PSP is specific to the stimulated synaptic pair (e.g., SN–MN1) and does not occur for the other pair (e.g., SN–MN2). These results should generalize to multiple pre- and postsynaptic partners and suggest two novel features of synaptic regulation during plasticity: (1) that the number of synaptic contacts between two neurons is regulated, and (2) that the regulation is homeostatic. We first describe those features and some of the evidence supporting them, then propose a model that could account for them and present computational modeling to illustrate the plausibility of the model.     

Insights into a molecular switch that gates sensory neuron synapses during habituation in Aplysia

This review focuses on synaptic depression at sensory neuron-to-motor neuron synapses in the defensive withdrawal circuit of Aplysia as a model system for analysis of molecular mechanisms of sensory gating and habituation. We address the following topics:1. Of various possible mechanisms that might underlie depression at these sensory neuron-to-motor neuron synapses in Aplysia, historically the most widely-accepted explanation has been depletion of the readily releasable pool of vesicles. Depletion is also believed to account for synaptic depression at long interstimulus intervals in a variety of other systems.2. Multiple lines of evidence now indicate that vesicle depletion is not an important contributing mechanism to synaptic depression at Aplysia sensory neuron-to-motor neuron synapses. More generally, it appears that vesicle depletion does not contribute substantially to depression that occurs with those stimulus patterns that are typically used in studying behavioral habituation.3. Recent evidence suggests that at these sensory neuron-to-motor neuron synapses in Aplysia, synaptic depression is mediated by an activity-dependent, but release-independent, switching of individual release sites to a silent state. This switching off of release sites is initiated by Ca²⁺ influx during individual action potentials. We discuss signaling proteins that may be regulated by Ca²⁺ during the silencing of release sites that underlies synaptic depression.4. Bursts of 2–4 action potentials in presynaptic sensory neurons in Aplysia prevent the switching off of release sites via a mechanism called “burst-dependent protection” from synaptic depression.5. This molecular switch may explain the sensory gating that allows animals to discriminate which stimuli are innocuous and appropriate to ignore and which stimuli are more important and should continue to elicit responses.     

α7 Nicotinic Receptor Subunits Are Not Necessary for Hippocampal-Dependent Learning or Sensorimotor Gating: A Behavioral Characterization of Acra7-Deficient Mice

The α7 nicotinic acetylcholine receptor (nAChR) subunit is abundantly expressed in the hippocampus and contributes to hippocampal cholinergic synaptic transmission suggesting that it may contribute to learning and memory. There is also evidence for an association between levels of α7 nAChR and in sensorimotor gating impairments. To examine the role of α7 nAChRs in learning and memory and sensorimotor gating, Acra7 homozygous mutant mice and their wild-type littermates were tested in a Pavlovian conditioned fear test, for spatial learning in the Morris water task, and in the prepulse inhibition paradigm. Exploratory activity, motor coordination, and startle habituation were also evaluated. Acra7 mutant mice displayed the same levels of contextual and auditory-cue condition fear as wild-type mice. Similarly, there were no differences in spatial learning performance between mutant and wild-type mice. Finally, Acra7 mutant and wild-type mice displayed similar levels of prepulse inhibition. Other behavioral responses in Acra7 mutant mice were also normal, except for an anxiety-related behavior in the open-field test. The results of this study show that the absence of α7 nAChRs has little impact on normal, base-line behavioral responses. Future studies will examine the contribution of α7 nAChR to the enhancement of learning and sensorimotor gating following nicotine treatments.     

Hyperactivity and Learning Deficits in Transgenic Mice Bearing a Human Mutant Thyroid Hormone β1 Receptor Gene

Resistance to thyroid hormone (RTH) is a human syndrome mapped to the thyroid receptor β (TRβ) gene on chromosome 3, representing a mutation of the ligandbinding domain of the TRβ gene. The syndrome is characterized by reduced tissue responsiveness to thyroid hormone and elevated serum levels of thyroid hormones. A common behavioral phenotype associated with RTH is attention deficit hyperactivity disorder (ADHD). To test the hypothesis that RTH produces attention deficits and/or hyperactivity, transgenic mice expressing a mutant TRβ gene were generated. The present experiment tested RTH transgenic mice from the PV kindred on behavioral tasks relevant to the primary features of ADHD: hyperactivity, sustained attention (vigilance), learning, and impulsivity. Male transgenic mice showed elevated locomotor activity in an open field compared to male wild-type littermate controls. Both male and female transgenic mice exhibited impaired learning of an autoshaping task, compared to wild-type controls. On a vigilance task in an operant chamber, there were no differences between transgenics and controls on the proportion of hits, response latency, or duration of stimulus tolerated. On an operant go/no-go task measuring sustained attention and impulsivity, there were no differences between controls and transgenics. These results indicate that transgenic mice bearing a mutant human TRβ gene demonstrate several behavioral characteristics of ADHD and may serve a valuable heuristic role in elucidating possible candidate genes in converging pathways for other causes of ADHD.     

Reversal of Relative Thresholds for Synaptic Facilitation and Increased Excitability Induced by Serotonin and Tail Nerve Stimulation inAplysiaSensory Neurons

Tail shock induces reflex sensitization inAplysiaand, in parallel, induces a number of modulatory effects in central neurons, such as increased excitability in tail sensory neurons (SNs) and facilitation of synaptic transmission from SNs to motor neurons. Both of these modulatory effects are mimicked by exogenous application of serotonin (5HT) or electrical stimulation of the tail nerve P9. In the present study we examined the activation thresholds for increased excitability and synaptic facilitation induced by either 5HT or P9 stimulation. We found that the concentration of 5HT sufficient to produce a significant increase in excitability produced no significant synaptic facilitation and, conversely, that the intensity of nerve stimulation sufficient to produce significant synaptic facilitation produced no excitability changes. This reversal of relative thresholds for these modulatory effects may reflect the differential access of exogenous 5HT and endogenous 5HT (released by tail nerve stimulation) to the SN cell body and synaptic terminals, respectively.     

Chronic nicotine exposure induces a long-lasting and pathway-specific facilitation of LTP in the amygdala

Nicotine, in the form of tobacco, is the most commonly used drug of abuse. In addition to its rewarding properties, nicotine also affects many cognitive and emotional processes that involve several brain regions, including hippocampus and amygdala. Long-term changes in synaptic strength in these brain regions after drug exposure may be importantly correlated with behavioral changes induced by nicotine. Here, we study the effect of chronic oral administration of nicotine on the long-term synaptic potentiation in the amygdala, a key structure for emotional memory. We find that oral administration of nicotine for 7 d produces a significant enhancement of LTP in the amygdala. This facilitation is pathway specific: Nicotine selectively facilitates LTP in the cortical-lateral amygdala pathway, but not the thalamic-lateral and the lateral-basolateral synaptic pathway. The synaptic facilitation induced by a 7-d exposure to nicotine is long-lasting, it persists for 72 h after cessation of nicotine but decays 8 d after its cessation. In contrast, a shorter exposure of nicotine (24 h) induces only a short-lasting facilitation of synaptic plasticity that dissipates 24 and 72 h after cessation of nicotine. The facilitation of LTP in the amygdala after exposure to nicotine is mediated by removal of GABAergic inhibition, is dependent on the activation NMDA receptors, and can be prevented by blocking either α7 or β2 nACh receptors. Our results indicate that chronic exposure to nicotine can promote the induction of long-lasting modifications of synapses in a specific pathway in the amygdala.These changes in synaptic plasticity may contribute to the complex neural adaptations and behaviors caused by nicotine.     

Inflammation Causes a Long-Term Hyperexcitability in the Nociceptive Sensory Neurons of Aplysia

Nerve injury, tissue damage, and inflammation all cause hyperalgesia. A factor contributing to this increased sensitivity is a long-term (>24 hr) hyperexcitability (LTH) in the sensory neurons that mediate the responses. Using the cluster of nociceptive sensory neurons in Aplysia californica as a model, we are examining how inflammation induces LTH. A general inflammatory response was induced by inserting a gauze pad into the animal. Within 4 days, the gauze is enmeshed in an amorphous material that contains hemocytes, which comprise a cellular immune system. Concurrently, LTH appears in both ipsilateral and contralateral sensory neurons. The LTH is manifest as increased action potential discharge to a normalized stimulus. Immunocytochemistry revealed that hemocytes have antigens recognized by antibodies to TGFβ1, IL-6, and 5HT. When a localized inflammation was elicited on a nerve, hemocytes containing the TGFβ1 antigen were present near axons within the nerve and those containing the IL-6 were on the surface. Western blots of hemocytes, or of gauze that had induced a foreign body response, contained a 28-kD polypeptide recognized by the anti-TGFβ1 antibody. Exposure of the nervous system to recombinant human TGFβ1 elicited increased firing of the nociceptive neurons and a decrease in threshold. The TGFβ1 also caused an activation of protein kinase C (PKC) in axons but did not affect a kinase that is activated in axons after injury. Our findings, in conjunction with previous results, indicate that a TGFβ1-homolog can modulate the activity of neurons that respond to noxious stimuli. This system could also contribute to interactions between the immune and nervous systems via regulation of PKC.     

Consolidation of Memory for Odor–Reward Association: β-Adrenergic Receptor Involvement in the Late Phase

Experimentally naive rats can learn rapidly to discriminate among three odors to obtain food reinforcement. After three massed trials, they show almost errorless performance. This task has proved to be useful in studying time-dependent postacquisition intracellular processes necessary for long-term memory. The present experiments evaluated the temporal dynamics of the role of β-noradrenergic receptors in long-term consolidation. Rats were implanted with intracerebroventricular cannulae and trained in a single session to find reinforcement in a hole in a sponge impregnated with a particular odor. Injections of the β-receptor antagonist timolol were made at 5 min, 1, 2, or 5 hr after training. Memory and relearning ability were evaluated 48 hr later. Rats treated with timolol 2 hr after training showed a memory deficit at the retention test, but were able to relearn the task normally. Injections at the earlier or later time points were ineffective. The results reinforce previous observations with systemic injections that β-noradrenergic receptors are involved in the late phase of memory consolidation and suggest a critical time window during which they are necessary. The time window is compatible with the current view that long-term memory depends on late involvement of the cAMP cascade leading to new protein synthesis necessary for synaptic reorganization.     

Lifelong reductions of PKMζ in ventral hippocampus of nonhuman primates exposed to early-life adversity due to unpredictable maternal care

Protein kinase Mζ (PKMζ) maintains long-term potentiation (LTP) and long-term memory through persistent increases in kinase expression. Early-life adversity is a precursor to adult mood and anxiety disorders, in part, through persistent disruption of emotional memory throughout life. Here we subjected 10- to 16-wk-old male bonnet macaques to adversity by a maternal variable-foraging demand paradigm. We then examined PKMζ expression in their ventral hippocampi as 7- to 12-yr-old adults. Quantitative immunohistochemistry reveals decreased PKMζ in dentate gyrus, CA1, and subiculum of subjects who had experienced early-life adversity due to the unpredictability of maternal care. Adult animals with persistent decrements of PKMζ in ventral hippocampus express timid rather than confrontational responses to a human intruder. Persistent down-regulation of PKMζ in the ventral hippocampus might reduce the capacity for emotional memory maintenance and contribute to the long-lasting emotional effects of early-life adversity.

Early-life adversity is associated with an increased vulnerability to stress-related disorders that is maintained into adulthood, suggesting a very long-lived effect on emotional memory by the early-life event (Coplan et al. 1996). Although several structural and neurochemical sequelae of early-life adversity have been reported (Teicher et al. 2003; Jackowski et al. 2011), the direct effects of early-life adversity on the molecular substrates maintaining long-term memory storage have not been explored.Accumulating evidence supports a crucial role for the autonomously active, atypical protein kinase C (PKC) isoform protein kinase Mζ (PKMζ) in maintaining synaptic long-term potentiation (LTP), a putative physical substrate for memory, and long-term memory storage (Ling et al. 2002; Pastalkova et al. 2006; Glanzman 2013; Sacktor and Fenton 2018). The autonomous activity of PKMζ is due to its unusual structure that differs from other PKC isoforms (Sacktor et al. 1993). Most PKCs consist of two domains: a catalytic domain and an autoinhibitory regulatory domain that suppresses the catalytic domain. Therefore, most PKCs are inactive until second messengers bind to the regulatory domain and induce a conformational change that releases the autoinhibition. Because second messengers that activate PKCs such as Ca²⁺ or diacylglycerol have short half-lives, most PKCs are only transiently activated.PKMζ, in contrast, consists of an independent PKCζ catalytic domain, and the absence of an autoinhibitory regulatory domain results in autonomous and thus persistent activity once the kinase is synthesized. PKMζ mRNA is transcribed from an internal promoter within the PKCζ/PKMζ gene that is active only in neural tissue (Hernandez et al. 2003). The mRNA is translationally repressed and transported to dendrites of neurons (Muslimov et al. 2004). High-frequency afferent synaptic activity during LTP induction or learning derepresses PKMζ mRNA translation, triggering new synthesis of PKMζ protein (Osten et al. 1996; Hernandez et al. 2003; Tsokas et al. 2016; Hsieh et al. 2017).Once increased, the steady-state amount of PKMζ remains elevated during LTP or long-term memory maintenance. Recent work with quantitative immunohistochemistry (IHC) shows that spatial conditioning induces persistent increases of PKMζ in somatic and selective dendritic compartments of dorsal hippocampal CA1 pyramidal cells that can last at least 1 mo (Hsieh et al. 2021). The persistent increases are preferentially expressed in CA1 pyramidal cells that were activated during the formation of the memory, specifically at the termination zone of the Schaffer collateral/commissural inputs from subfield CA3. In contrast, persistent PKMζ increases are not evident in stratum lacunosum-moleculare, the termination zone that originates in entorhinal cortex that nonetheless is capable of expressing PKMζ. Postsynaptic domain-specific PKMζ expression patterns hint at distinct circuit-specific modifications of cortical–hippocampal synaptic function by maturational and experiential factors.Persistent changes in PKMζ expression are also associated with changes in the capacity for learning and memory across the life span of animals. Decreased memory ability in aged rats is associated with decreased training-induced, persistent PKMζ expression in prelimbic cortex, and increases in PKMζ are crucial for the cognition-enhancing effects of environmental enrichment in the aged animals (Chen et al. 2016). Hara et al. extended the connection between PKMζ and cognitive function to nonhuman primates (NHPs), showing that levels of PKMζ expression in dentate gyrus (DG) axospinous synapses correlate with successful performance on cognitive tasks in young and aged monkeys (Hara et al. 2012). These studies suggest that persistent down-regulation of PKMζ may comprise an important pathophysiological mechanism for cognitive impairment.Here we used a validated NHP model of early-life adversity, maternal variable-foraging demand (VFD), to explore the links between adversity in infancy and PKMζ expression in adulthood (Coplan et al. 1996; Jackowski et al. 2011). Previous studies of the VFD paradigm have revealed that both infants and their mothers exposed to VFD show significant cerebrospinal fluid (CSF) elevations of the stress neuropeptide, corticotropin-releasing factor (CRF). Moreover, the magnitude of CRF change in mothers and infants are positively correlated, suggesting synchronization of maternal–infant stress responses to the VFD stressor (Coplan et al. 2005). From a behavioral standpoint, maternal social rank plays a negligible role in determining an aggregate score of maternal–infant proximity, suggesting preferential attention of mothers to their infants. During the VFD condition, maternal social rank predicts >80% of the variance of maternal–infant proximity, suggesting mothering patterns are interrupted by preferential orientations to social rank; the latter determines food accessibility (Coplan et al. 2015). Dominant females show relative increases in maternal–infant proximity, whereas subordinate females show relative reductions in maternal–infant proximity. Neither pattern of attachment ameliorates an abnormal association between CSF oxytocin concentrations and hypothalamic-pituitary-adrenal (HPA) axis activity (Coplan et al. 2015). Offspring exposed to VFD rearing assessed both as juveniles and as full adults demonstrate persistent increases in CSF CRF concentrations in comparison with controls reared under non-VFD conditions (Coplan et al. 1996, 2001).Our prior neurohistological studies pointed to the DG as a region particularly vulnerable to VFD exposure, as shown by reduced trophic signaling and neurogenesis (Jackowski et al. 2011; Perera et al. 2011; Schoenfeld et al. 2021). We therefore hypothesized that early-life adversity due to unpredictable maternal care (for brevity, subsequently referred to as “early-life adversity”) reduces the persistent expression of PKMζ within the DG of ventral intrahippocampal neurocircuitry that mediates affective memory processing (Fanselow and Dong 2010). We used PKMζ antisera validated by the lack of immunostaining in PKMζ-null mice (Hsieh et al. 2021) to examine PKMζ expression in ventral hippocampus (NHP anterior hippocampus) in both DG granule cell layer and the stratum moleculare of the suprapyramidal blade that receives direct input from entorhinal cortex, as well as other regions encompassing the hippocampal formation, including the hilus, CA3, CA1, and subiculum.To assess behavioral correlates of hippocampal PKMζ expression, we used a stress-inducing paradigm designed specifically for singly housed bonnet macaque male NHPs, which we refer to as the “human exposure response” (Jackowski et al. 2011; Hamel et al. 2017), which is a variation of the paradigm used in human exposure studies by Kalin et al. in rhesus macaques (Kalin and Shelton 1989). On exposure to a direct human presence, singly housed adult male bonnet macaques react with a dichotomy of responses—confrontational versus timid (see the Materials and Methods) (Jackowski et al. 2011). In our macaque colony, groups of fully adult males are necessarily housed individually to prevent injury sustained during male agonistic encounters, whereas adult females and/or juveniles are safely housed in social groups. Because group housing of nursing females and/or juveniles of both sexes elicits a range of behaviors intrinsic to the species’ social repertoire (Rosenblum et al. 2001; Coplan et al. 2015) that complicates behavioral analyses to human exposure, we restricted our current study to male macaques.     

Neuronal NT-3 Is not Required For Synaptic Transmission or Long-Term Potentiation in Area CA1 of the Adult Rat Hippocampus

Neurotrophic factors, including BDNF and NT-3, have been implicated in the regulation of synaptic transmission and plasticity. Previous attempts to analyze synaptic transmission and plasticity in mice lacking the NT-3 gene have been hampered by the early death of the NT-3 homozygous knockout animals. We have bypassed this problem by examining synaptic transmission in mice in which the NT-3 gene is deleted in neurons later in development, by crossing animals expressing the CRE recombinase driven by the synapsin I promoter to animals in which the NT-3 gene is floxed. We conducted blind field potential recordings at the Schaffer collateral–CA1 synapse in hippocampal slices from homozygous knockout and wild-type mice. We examined the following indices of synaptic transmission: (1) input-output relationship; (2) paired-pulse facilitation; (3) post-tetanic potentiation; and (4) long-term potentiation: induced by two different protocols: (a) two trains of 100-Hz stimulation and (b) theta burst stimulation. We found no difference between the knockout and wild-type mice in any of the above measurements. These results suggest that neuronal NT-3 does not play an essential role in normal synaptic transmission and some forms of plasticity in the mouse hippocampus.     

Habituation in Aplysia: The Cheshire Cat of neurobiology

The marine snail, Aplysia californica, is a valuable model system for cell biological studies of learning and memory. Aplysia exhibits a reflexive withdrawal of its gill and siphon in response to weak or moderate tactile stimulation of its skin. Repeated tactile stimulation causes this defensive withdrawal reflex to habituate. Both short-term habituation, lasting <30 min, and long-term habituation, which can last >24 h, have been reported in Aplysia. Habituation of the withdrawal reflex correlates with, and is in part due to, depression of transmission at the monosynaptic connection between mechanoreceptive sensory neurons and motor neurons within the abdominal ganglion. Habituation-related short-term depression of the sensorimotor synapse appears to be due exclusively to presynaptic changes. However, changes within the sensory neuron, by themselves, do not account for more persistent depression of the sensorimotor synapse. Recent behavioral work suggests that long-term habituation in Aplysia critically involves postsynaptic processes, specifically, activation of AMPA- and NMDA-type receptors. In addition, long-term habituation requires activity of protein phosphatases, including protein phosphatases 1, 2A, and 2B, as well as activity of voltage-dependent Ca²⁺ channels. Cellular work has succeeded in demonstrating long-term, homosynaptic depression (LTD) of the sensorimotor synapse in dissociated cell culture and, more recently, LTD of the glutamate response of isolated motor neurons in culture (“hemisynaptic” LTD). These in vitro forms of LTD have mechanistic parallels to long-term habituation. In particular, homosynaptic LTD of the sensorimotor synapse requires elevated intracellular Ca²⁺ within the motor neuron, and hemisynaptic LTD requires activity of AMPA- and NMDA-type receptors. In addition, activation of group I and II metabotropic glutamate receptors (mGluRs) can induce hemisynaptic LTD. The demonstration of LTD in vitro opens up a promising new avenue for attempts to relate long-term habituation to cellular changes within the nervous system of Aplysia.     

Synaptic Reliability Correlates with Reduced Susceptibility to Synaptic Potentiation by Brain-Derived Neurotrophic Factor

Recent studies have implicated brain-derived neurotrophic factor (BDNF) in use-dependent modification of hippocampal synapses. BDNF can rapidly potentiate synaptic transmission at glutamatergic synapses by enhancing transmitter release. Using simultaneous perforated patch recording from pairs and triplets of glutamatergic hippocampal neurons, we have examined how the initial state of the glutamatergic synapse determines its susceptibility to synaptic modification by BDNF. We found that the degree of synaptic potentiation by BDNF depends on the initial reliability and strength of the synapse: Relatively weak connections were strongly potentiated, whereas the effect was markedly reduced at stronger synapses. The degree of BDNF-induced potentiation strongly correlated with the initial coefficient of variation (CV) of the amplitude of excitatory postsynaptic currents (EPSCs) and inversely correlated with the initial paired–pulse facilitation, suggesting that synapses with lower release probability (P_r) are more susceptible to the action of BDNF. To determine whether saturation of P_r could have masked the potentiation effect of BDNF in the stronger synapses, we lowered the initial P_r either by reducing the extracellular Ca²⁺ concentration ([Ca²⁺]_o) or by bath application of adenosine. Synapses that were initially strong remained unaffected by BDNF under these conditions of reduced P_r. Thus, the lack of BDNF effect on synaptic efficacy cannot simply be accounted for by saturation of P_r, but rather may be due to intrinsic changes associated with synaptic maturation that might covary with P_r. Finally, the dependence on initial synaptic strength was also found for divergent outputs of the same presynaptic neuron, suggesting that synaptic terminals with different degrees of responsiveness to BDNF can coexist within in the same neuron.     

Transgenic Brain-Derived Neurotrophic Factor Modulates a Developing Cerebellar Inhibitory Synapse

Brain-derived neurotrophic factor (BDNF) has been shown to promote synapse formation and maturation in neurons of many brain regions, including inhibitory synapses. In the cerebellum, the Golgi cell-granule cell GABAergic synaptic responses undergo developmental transition from slow-decaying to fast-decaying kinetics, which parallels a developmental increase of GABA_A receptor α6 subunit expression in the cerebellar granule cells. In culture, BDNF accelerates the expression of GABA_A receptor α6 subunit expression in granule cells. Here we examined synaptic GABA_A response kinetics in BDNF transgenic mice. The mutant mouse, which carries a BDNF transgene driven by a β-actin promoter, overexpresses BDNF (two- to fivefold increase compared with wild types) in all brain regions. Recordings of the spontaneous GABA_A responses indicate that the decay time constant of the GABAergic responses decreases during early postnatal development; this transition is accelerated in the BDNF transgenic mouse. The amplitude of the spontaneous GABA_A responses was also larger in the transgenic mouse than in the wild-type mouse. However, the frequency of the spontaneous GABA_A responses were not different between the two groups. Our results suggest that BDNF may modulate GABAergic synapse maturation in the cerebellum.     

Corticosterone time-dependently modulates beta-adrenergic effects on long-term potentiation in the hippocampal dentate gyrus

Previous experiments in the hippocampal CA1 area have shown that corticosterone can facilitate long-term potentiation (LTP) in a rapid non-genomic fashion, while the same hormone suppresses LTP that is induced several hours after hormone application. Here, we elaborated on this finding by examining whether corticosterone exerts opposite effects on LTP depending on the timing of hormone application in the dentate gyrus as well. Moreover, we tested rapid and delayed actions by corticosterone on β-adrenergic-dependent changes in LTP. Unlike the CA1 region, our in vitro field potential recordings show that rapid effects of corticosterone do not influence LTP induced by mild tetanization in the hippocampal dentate gyrus, unless GABA_A receptors are blocked. In contrast, the β-adrenergic agonist isoproterenol does initiate a slow-onset, limited amount of potentiation. When corticosterone was applied concurrently with isoproterenol, a further enhancement of synaptic strength was identified, especially during the early stage of potentiation. Yet, treatment with corticosterone several hours in advance of isoproterenol fully prevented any effect of isoproterenol on LTP. This emphasizes that corticosterone can regulate β-adrenergic modulation of synaptic plasticity in opposite directions, depending on the timing of hormone application.