Sex-dependent differences in the ability of nicotine to modulate discrimination learning and cognitive flexibility in mice.

Nicotine, an addictive compound found in tobacco, functions as an agonist of nicotinic acetylcholine receptors (nAChRs) in the brain. Interestingly, nicotine has been reported to act as a cognitive enhancer in both human subjects and experimental animals. However, its effects in animal studies have not always been consistent, and sex differences have been identified in the effects of nicotine on several behaviors. Specifically, the role that sex plays in modulating the effects of nicotine on discrimination learning and cognitive flexibility in rodents is still unclear. Here, we evaluated sex-dependent differences in the effect of daily nicotine intraperitoneal (i.p.) administration at various doses (0.125, 0.25, and 0.5 mg/kg) on visual discrimination (VD) learning and reversal (VDR) learning in mice. In male mice, 0.5 mg/kg nicotine significantly improved performance in the VDR, but not the VD, task, while 0.5 mg/kg nicotine significantly worsened performance in the VD, but not VDR task in female mice. Furthermore, 0.25 mg/kg nicotine significantly worsened performance in the VD and VDR task only in female mice. Next, to investigate the cellular mechanisms that underlie the sex difference in the effects of nicotine on cognition, transcriptomic analyses were performed focusing on the medial prefrontal cortex tissue samples from male and female mice that had received continuous administration of nicotine for 3 or 18 days. As a result of pathway enrichment analysis and protein-protein interaction analysis using gene sets of differentially expressed genes, decreased expression of postsynaptic-related genes in males and increased expression of innate immunity-related genes in females were identified as possible molecular mechanisms related to sex differences in the effects of nicotine on cognition in discrimination learning and cognitive flexibility. Our result suggests that nicotine modulates cognitive function in a sex-dependent manner by alternating the expression of specific gene sets in the medial prefrontal cortex.

Altered Heterosynaptic Plasticity Impairs Visual Discrimination Learning in Adenosine A1 Receptor Knock-Out Mice.

Theoretical and modeling studies demonstrate that heterosynaptic plasticity-changes at synapses inactive during induction-facilitates fine-grained discriminative learning in Hebbian-type systems, and helps to achieve a robust ability for repetitive learning. A dearth of tools for selective manipulation has hindered experimental analysis of the proposed role of heterosynaptic plasticity in behavior. Here we circumvent this obstacle by testing specific predictions about the behavioral consequences of the impairment of heterosynaptic plasticity by experimental manipulations to adenosine A1 receptors (A1Rs). Our prior work demonstrated that the blockade of adenosine A1 receptors impairs heterosynaptic plasticity in brain slices and, when implemented in computer models, selectively impairs repetitive learning on sequential tasks. Based on this work, we predict that A1R knock-out (KO) mice will express (1) impairment of heterosynaptic plasticity and (2) behavioral deficits in learning on sequential tasks. Using electrophysiological experiments in slices and behavioral testing of animals of both sexes, we show that, compared with wild-type controls, A1R KO mice have impaired synaptic plasticity in visual cortex neurons, coupled with significant deficits in visual discrimination learning. Deficits in A1R knockouts were seen specifically during relearning, becoming progressively more apparent with learning on sequential visual discrimination tasks of increasing complexity. These behavioral results confirm our model predictions and provide the first experimental evidence for a proposed role of heterosynaptic plasticity in organism-level learning. Moreover, these results identify heterosynaptic plasticity as a new potential target for interventions that may help to enhance new learning on a background of existing memories.SIGNIFICANCE STATEMENT Understanding how interacting forms of synaptic plasticity mediate learning is fundamental for neuroscience. Theory and modeling revealed that, in addition to Hebbian-type associative plasticity, heterosynaptic changes at synapses that were not active during induction are necessary for stable system operation and fine-grained discrimination learning. However, lacking tools for selective manipulation prevented behavioral analysis of heterosynaptic plasticity. Here we circumvent this barrier: from our prior experimental and computational work we predict differential behavioral consequences of the impairment of Hebbian-type versus heterosynaptic plasticity. We show that, in adenosine A1 receptor knock-out mice, impaired synaptic plasticity in visual cortex neurons is coupled with specific deficits in learning sequential, increasingly complex visual discrimination tasks. This provides the first evidence linking heterosynaptic plasticity to organism-level learning.

The underestimated giants: operant conditioning, visual discrimination and long-term memory in giant tortoises.

Relatively little is known about cognition in turtles, and most studies have focused on aquatic animals. Almost nothing is known about the giant land tortoises. These are visual animals that travel large distances in the wild, interact with each other and with their environment, and live extremely long lives. Here, we show that Galapagos and Seychelle tortoises, housed in a zoo environment, readily underwent operant conditioning and we provide evidence that they learned faster when trained in the presence of a group rather than individually. The animals readily learned to distinguish colors in a two-choice discrimination task. However, since each animal was assigned its own individual colour for this task, the presence of the group had no obvious effect on the speed of learning. When tested 95 days after the initial training, all animals remembered the operant task. When tested in the discrimination task, most animals relearned the task up to three times faster than naïve animals. Remarkably, animals that were tested 9 years after the initial training still retained the operant conditioning. As animals remembered the operant task, but needed to relearn the discrimination task constitutes the first evidence for a differentiation between implicit and explicit memory in tortoises. Our study is a first step towards a wider appreciation of the cognitive abilities of these unique animals.

Neutral point testing of color vision in the domestic cat.

Despite extensive study, the basic nature of feline spectral sensitivity is still unresolved. Most electrophysiological studies have demonstrated two photopic receptors within the cat's retina, one most sensitive to longer wavelengths near 560 nm and the other most sensitive to shorter wavelengths near 460 nm, providing the neuroretinal basis for dichromatic vision. A few studies, however, have detected a third photopic receptor most sensitive to medium wavelengths between 500 and 520 nm, overlapping in spectrally sensitivity with the feline scotopic receptor, that potentially could allow trichromatic vision. Indeed, one behavioral study has demonstrated trichromatic vision in cats, but a flaw within its experimental design raises the possibility that achromatic intensity cues might have allowed the accurate identification of medium wavelength targets. This study tested for a spectral neutral point in the domestic cat using a two-choice discrimination task. The positive targets were created using monochromatic light from various single wavelength light emitting diodes (LEDs) combined with a white light of variable intensity, while the negative targets were created using white light of variable intensity. Trials were performed with varying intensities of positive and negative targets, from brighter positive targets to brighter negative targets, to eliminate achromatic intensity cues. Two cats with prior experience with two-choice discrimination tasks, one male and one female, successfully discriminated monochromatic light from 456 nm to 497 nm and from 510 nm to 524 nm, but both failed to discriminate monochromatic light at 505 nm over multiple trials. These results provide strong evidence that cats are dichromatic with a neutral point near 505 nm. This neutral point is nearly identical to the neutral point of the human deuteuranope, making feline vision a more accurate a model for red-green colorblind individuals than normal trichromats.

Dynamic functional brain networks involved in simple visual discrimination learning.

Visual discrimination tasks have been widely used to evaluate many types of learning and memory processes. However, little is known about the brain regions involved at different stages of visual discrimination learning. We used cytochrome c oxidase histochemistry to evaluate changes in regional brain oxidative metabolism during visual discrimination learning in a water-T maze at different time points during training. As compared with control groups, the results of the present study reveal the gradual activation of cortical (prefrontal and temporal cortices) and subcortical brain regions (including the striatum and the hippocampus) associated to the mastery of a simple visual discrimination task. On the other hand, the brain regions involved and their functional interactions changed progressively over days of training. Regions associated with novelty, emotion, visuo-spatial orientation and motor aspects of the behavioral task seem to be relevant during the earlier phase of training, whereas a brain network comprising the prefrontal cortex was found along the whole learning process. This study highlights the relevance of functional interactions among brain regions to investigate learning and memory processes.

The effects of time, luminance, and high contrast targets: revisiting grating acuity in the domestic cat.

Based on optical clarity and retinal cone density, the cat has a potential acuity of 20-30 cycles per degree (cpd), yet most behavioral studies estimate feline acuity between 3 and 9 cpd. Those studies, however, were limited by restrictive experimental conditions that may have inadvertently lowered the estimated grating acuity. Two domestic cats previously trained on a two-choice visual discrimination task were retrained on a grating detection/discrimination task with unlimited time, high luminance, high contrast targets, and adequate space to prevent poor accommodation from affecting the results. Initially, vertical gratings of increasing cpd were tested until failure. Then, horizontal gratings of increasing cpd were tested until failure. Finally, the finest horizontal grating resolved was confirmed with a third test requiring 24 correct out of 36 consecutive trials, yielding a binomial probability less than 0.02 of non-random occurrence. M1, a 7-year-old male gray tabby with +2.00 OU refraction, tested for a grating detection acuity of 15 cpd for both vertical and horizontal gratings (binomial probability = 0.009). F1, a 2-year-old female gray tabby with +0.25 OU refraction, tested for a grating orientation discrimination acuity of 20 cpd for both vertical and horizontal gratings (binomial probability = 0.004). These results demonstrate that a young cat with good focus is capable of discriminating 20 cpd, in close agreement with the physiologic maximum. Uncorrected focusing errors appear to degrade visual performance. Optimum experimental conditions resulted in better grating acuity measurements than previously reported, emphasizing the importance of environmental factors in feline behavioral testing.

Relationship between hippocampal gene expression and cognitive performance differences in visual discrimination learning task of male rats.

Learning to discriminate between environmental visual stimuli is essential to make right decisions and guide appropriate behaviors. Moreover, impairments in visual discrimination learning are observed in several neuropsychiatric disorders. Visual discrimination learning requires perception and memory processing, in which the hippocampus critically involved. To understand the molecular mechanisms underpinning hippocampus function in visual discrimination learning, we examined the hippocampal gene expression profiles of Sprague-Dawley rats with different cognitive performance (high cognition group vs. low cognition group) in the modified visual discrimination learning task, using high-throughput RNA sequencing technology. Compared with the low cognition group, bioinformatics analysis indicated that 319 genes were differentially expressed in the high cognition group with statistical significance, of which 253 genes were down-regulated and 66 genes were up-regulated. The functional enrichment analysis showed that protein translation and energy metabolism were up-regulated pathways, while transforming growth factor beta receptor signaling pathway, bone morphogenetic protein signaling pathway, apoptosis, inflammation response, transport, and glycosaminoglycan metabolism were down-regulated pathways, which were related to good cognitive performance in the visual discrimination learning task. Taken together, our finding reveals the differential gene expression and enrichment biological pathways related to cognitive performance differences in visual discrimination learning of rats, which provides us direct insight into the molecular mechanisms of hippocampus function in visual discrimination learning and may contribute to developing potential treatment strategies for neuropsychiatric disorders accompanied with cognitive impairments.

Visual discrimination and reversal learning in aged common marmosets (Callithrix jacchus).

Common marmosets (Callithrix jacchus) have been suggested as a new model for analysis of age-related changes and neurodegenerative diseases. However, the effects of age on learning and memory processes are not well defined within this species. Therefore, we employed visual discrimination and reversal learning tasks to evaluate learning and memory in four aged common marmosets relative to a younger cohort. We found that aged marmosets commit significantly more errors in initial stages of visual discrimination and more perseverative errors in reversal learning, indicating prefrontal dysfunction. However, they showed comparable performance with younger marmosets in the later stages of both tasks.

Characteristic and intermingled neocortical circuits encode different visual object discriminations.

Synaptic plasticity and neural network theories hypothesize that the essential information for advanced cognitive tasks is encoded in specific circuits and neurons within distributed neocortical networks. However, these circuits are incompletely characterized, and we do not know if a specific discrimination is encoded in characteristic circuits among multiple animals. Here, we determined the spatial distribution of active neurons for a circuit that encodes some of the essential information for a cognitive task. We genetically activated protein kinase C pathways in several hundred spatially-grouped glutamatergic and GABAergic neurons in rat postrhinal cortex, a multimodal associative area that is part of a distributed circuit that encodes visual object discriminations. We previously established that this intervention enhances accuracy for specific discriminations. Moreover, the genetically-modified, local circuit in POR cortex encodes some of the essential information, and this local circuit is preferentially activated during performance, as shown by activity-dependent gene imaging. Here, we mapped the positions of the active neurons, which revealed that two image sets are encoded in characteristic and different circuits. While characteristic circuits are known to process sensory information, in sensory areas, this is the first demonstration that characteristic circuits encode specific discriminations, in a multimodal associative area. Further, the circuits encoding the two image sets are intermingled, and likely overlapping, enabling efficient encoding. Consistent with reconsolidation theories, intermingled and overlapping encoding could facilitate formation of associations between related discriminations, including visually similar discriminations or discriminations learned at the same time or place.

Stagewise multidimensional visual discrimination by pigeons.

We trained six pigeons in a stagewise Multiple Necessary Cues (MNC) go/no-go task to document the dynamics of discrimination learning involving increasingly complex visual stimuli. The compound stimuli were composed from four dimensions, each of which could assume either of two extreme values or their intermediate value: Shape, Size, Line Orientation, and Brightness. Starting with a stimulus composed entirely from intermediate values, we replaced those values with each of the two extreme dimensional values in four successive stages, thereby increasing the stimulus set from 2 in Stage 1 to 16 in Stage 4. In each stage, only one combination of values signaled food (S+ ), whereas the remaining combinations did not (S- s). We calculated the rate of pecking during the first 15 s of each stimulus presentation and, in any given stage, training continued until the rate of responding to all of the S- s was less than 20% of the rate of responding to the S+ . All pigeons successfully acquired the final discrimination, suggesting that they attended to all of the dimensions relevant for the discrimination. We also replicated the key results of prior MNC studies: (1) the number of extreme dimensional values in each stage was positively related to the amount of training required for pigeons to acquire the discrimination; (2) attentional tradeoffs were most often observed when three or four dimensions were being trained; and (3) throughout training, the number of dimensional values in the S- s that differed from the S+ was positively related to their discriminability from S+ .

Individual variability in visual discrimination and reversal learning performance in common marmosets.

Detailed information about the characteristics of learning behavior in marmosets is useful for future marmoset research. We trained 42 marmosets in visual discrimination and reversal learning. All marmosets could learn visual discrimination, and all but one could complete reversal learning, though some marmosets failed to touch the visual stimuli and were screened out. In 87% of measurements, the final percentage of correct responses was over 95%. We quantified performance with two measures: onset trial and dynamic interval. Onset trial represents the number of trials that elapsed before the marmoset started to learn. Dynamic interval represents the number of trials from the start before reaching the final percentage of correct responses. Both measures decreased drastically as a result of the formation of discrimination learning sets. In reversal learning, both measures worsened, but the effect on onset trial was far greater. The effects of age and sex were not significant as far as we used adolescent or young adult marmosets. Unexpectedly, experimental circumstance (in the colony or isolator) had only a subtle effect on performance. However, we found that marmosets from different families exhibited different learning process characteristics, suggesting some family effect on learning.