Sex differences in avoidance behavior after perceiving potential risk in mice.

BACKGROUND: Sex has been considered as a potential factor regulating individual behaviors in different contexts. Recently, findings on sex differences in the neuroendocrine circuit have expanded due to exact measurements and control of neuronal activity, while findings on sex differences in behavioral phenotypes are limited. One efficient way to determine the miscellaneous aspects of a sexually different behavior is to segment it into a set of simpler responses induced by discrete scenes. METHODS: In the present study, we conducted a battery of behavioral tests within a variety of unique risky scenes, to determine where and how sex differences arise in responses under those scenes. RESULTS: A significant sex difference was observed in the avoidance responses measured in the two-way active and the passive avoidance tests. The phenotype observed was higher mobility in male mice and reduced mobility in female mice, and required associative learning between an escapable risk and its predictive cue. This was limited in other scenes where escapable risk or predictive cue or both were missing. CONCLUSIONS: Taken together, the present study found that the primary sex difference occurs in mobility in the avoidance response after perceiving escapable risks.

The effect of conversation on altruism: A comparative study with different media and generations.

Despite the overwhelming evidence of climate change and its effects on future generations, most individuals are still hesitant to make environmental changes that would especially benefit future generations. In this study, we investigate whether dialogue can influence people's altruistic behavior toward future generations of humans, and how it may be affected by participant age and the appearance of the conversation partner. We used a human, an android robot called Telenoid, and a speaker as representatives of future generations. Participants were split among an old age group and a young age group and were randomly assigned to converse with one of the aforementioned representatives. We asked the participants to play a round of the Dictator Game with the representative they were assigned, followed by an interactive conversation and another round of the Dictator Game in order to gauge their level of altruism. The results show that, on average, participants gave more money after having an interactive conversation, and that older adults tend to give more money than young adults. There were no significant differences between the three representatives. The results show that empathy might have been the most important factor in the increase in altruistic behavior for all participants.

DOPAnization of tyrosine in α-synuclein by tyrosine hydroxylase leads to the formation of oligomers.

Parkinson's disease is a progressive neurodegenerative disorder characterized by the preferential loss of tyrosine hydroxylase (TH)-expressing dopaminergic neurons in the substantia nigra. Although the abnormal accumulation and aggregation of α-synuclein have been implicated in the pathogenesis of Parkinson's disease, the underlying mechanisms remain largely elusive. Here, we found that TH converts Tyr136 in α-synuclein into dihydroxyphenylalanine (DOPA; Y136DOPA) through mass spectrometric analysis. Y136DOPA modification was clearly detected by a specific antibody in the dopaminergic neurons of α-synuclein-overexpressing mice as well as human α-synucleinopathies. Furthermore, dopanized α-synuclein tended to form oligomers rather than large fibril aggregates and significantly enhanced neurotoxicity. Our findings suggest that the dopanization of α-synuclein by TH may contribute to oligomer and/or seed formation causing neurodegeneration with the potential to shed light on the pathogenesis of Parkinson's disease.

Postsynaptic excitability is necessary for strengthening of cortical sensory responses during experience-dependent development.

Sensory experience is necessary for normal cortical development. This has been shown by sensory deprivation and pharmacological perturbation of the cortex. Because these manipulations affect the cortical network as a whole, the role of postsynaptic cellular properties during experience-dependent development is unclear. Here we addressed the developmental role of somatodendritic excitability, which enables postsynaptic spike timing-dependent forms of plasticity, in rat somatosensory cortex. We used short interfering RNA (siRNA)-based knockdown of Na+ channels to suppress the somatodendritic excitability of small numbers of layer 2/3 pyramidal neurons in the barrel cortex, without altering the ascending sensory pathway. In vivo recordings from siRNA-expressing cells revealed that this manipulation interfered with the normal developmental strengthening of sensory responses. The sensory responsiveness of neighboring cortical neurons was unchanged, indicating that the cortical network was unchanged. We conclude that somatodendritic excitability of the postsynaptic neuron is needed for the regulation of synaptic strength in the developing sensory cortex.

Acute stress increases neuropsin mRNA expression in the mouse hippocampus through the glucocorticoid pathway.

Stress affects synaptic plasticity and may alter various types of behaviour, including anxiety or memory formation. In the present study, we examined the effects of acute stress (1 h restraint with or without tail-shock) on mRNA levels of a plasticity-related serine protease neuropsin (NP) in the hippocampus using semiquantitative RT-PCR and in situ hybridization. We found that NP mRNA expression was dramatically increased shortly after exposure to the acute restraint tail-shock stress and remained at high level for at least 24 h. The level of NP mRNA would be correlated to the elevated plasma concentration of the glucocorticoid corticosterone (CORT) and to the stress intensity. Application of CORT either onto primary cultured hippocampal neurons (5 nM) or in vivo to adrenalectomized (ADX) mice (10 mg/kg B.W., s.c.) mimicked the effect of stress and significantly elevated NP mRNA. These results suggest that the upregulation of NP mRNA after stress is CORT-dependent and point to a role for neuropsin in stress-induced neuronal plasticity.

Auditory-induced response in the primary sensory cortex of rodents.

The details of auditory response at the subthreshold level in the rodent primary somatosensory cortex, the barrel cortex, have not been studied extensively, although several phenomenological reports have been published. Multisensory features may act as neuronal representations of links between inputs from one sensory modality to other sensory modalities. Here, we examined the basic multisensory postsynaptic responses in the rodent barrel cortex using in vivo whole-cell recordings of neurons. We observed robust responses to acoustic stimuli in most barrel cortex neurons. Acoustically evoked responses were mediated by hearing and reached approximately 60% of the postsynaptic response amplitude elicited by strong somatosensory stimuli. Compared to tactile stimuli, auditory stimuli evoked postsynaptic potentials with a longer latency and longer duration. Specifically, auditory stimuli in barrel cortex neurons appeared to trigger "up states", episodes associated with membrane depolarization and increased synaptic activity. Taken together, our data suggest that barrel cortex neurons have multisensory properties, with distinct synaptic mechanisms underlying tactile and non-tactile responses.

Enriched environment and Mash1 transfection affect neural stem cell differentiation after transplantation into the adult somatosensory cortex.

Neural stem cell (NSC) transplantation is a promising therapeutic modality for various nervous-system disorders; however, poor survival and differentiation of the transplanted NSCs limit their therapeutic efficacy. This study elucidated the effect of additive rehabilitative therapy with enriched environment (EE) and of achaete-scute homolog 1 (Mash1) and neurogenin2 (Ngn2) transduction on the fate of NSCs (P28-P35) transplanted into the primary somatosensory cortex (PSC) of mice. NSCs transplanted into the PSC differentiated into neurons and astrocytes and exhibited typical excitatory and synaptic response in mice housed in standard cages or in the EE. After EE exposure, significantly enhanced differentiation of transplanted NSCs into neuronal nuclear antigen-positive neurons was observed, whereas marked inhibition of the differentiation of transplanted NSCs into astrocytes was noted. Additionally, the proportion of GAD+ cells among GFP+/NeuN+ cells decreased following EE exposure. Furthermore, Mash1-transduced NSCs exhibited significantly enhanced populations of glutamic acid decarboxylase-negative neurons, whereas Ngn2-transduced NPCs did not.

Age-dependent enhancement of hippocampal long-term potentiation and impairment of spatial learning through the Rho-associated kinase pathway in protein tyrosine phosphatase receptor type Z-deficient mice.

Although protein tyrosine phosphatases (PTPs) are expressed abundantly in the brain, their roles in synaptic plasticity have not been well elucidated. In this study, we have examined the physiological functions of Ptprz, which is a receptor-type PTP expressed predominantly in the brain as a chondroitin sulfate proteoglycan. We have examined phenotypes of mutant mice deficient in Ptprz using electrophysiological, pharmacological, and behavioral approaches. Mutant mice exhibit enhanced long-term potentiation (LTP) in the CA1 region of hippocampal slices and impaired spatial learning abilities in an age-dependent manner: young adult (<10 weeks old) mutant mice show normal LTP and learning abilities in the Morris water maze task, whereas adult (>13 weeks old) mutant mice exhibit enhanced LTP and impairment in the task. The enhanced LTP is specifically canceled out by pharmacological inhibition of Rho-associated kinase (ROCK), a major downstream effector of Rho. These findings suggest that the lack of Ptprz leads to aberrant activation of ROCK and resultantly to enhanced LTP in the slice and learning impairments in the animal.

Mouse Ability to Perceive Subjective Contours.

In contrast to the previously held notion that mice have a weak visual system, it is now generally accepted that mice can perceive rather complicated figures in various contexts such as in cognitive experiments and in social settings. Here, we show that mice could even be capable of perceiving a visual illusion--subjective contours. This illusion requires the visual system to compensate for a lack of visual information in compressed 2D images on the retina. In this experiment, we trained mice to respond appropriately to a rectangle-shaped rewarded figure of specific orientation in a two-choice visual discrimination task with a touchscreen monitor. In Transfer Test 1, mice could discriminate illusory rectangle-shaped figures significantly as compared with a figure, which did not induce illusory figures. In Transfer Test 2, the choice rate of targets decreased with imperfect illusory figures, which produced weak perception of rotated or deficient inducers. Moreover, in Transfer Test 3, mice could not discriminate the low-resolution illusory figure, which also induced weak perception. These results demonstrated the possibility that mice might be useful for investigating fundamental properties of the neural visual system.

Conditional Spike Transmission Mediated by Electrical Coupling Ensures Millisecond Precision-Correlated Activity among Interneurons In Vivo.

Many GABAergic interneurons are electrically coupled and in vitro can display correlated activity with millisecond precision. However, the mechanisms underlying correlated activity between interneurons in vivo are unknown. Using dual patch-clamp recordings in vivo, we reveal that in the presence of spontaneous background synaptic activity, electrically coupled cerebellar Golgi cells exhibit robust millisecond precision-correlated activity which is enhanced by sensory stimulation. This precisely correlated activity results from the cooperative action of two mechanisms. First, electrical coupling ensures slow subthreshold membrane potential correlations by equalizing membrane potential fluctuations, such that coupled neurons tend to approach action potential threshold together. Second, fast spike-triggered spikelets transmitted through gap junctions conditionally trigger postjunctional spikes, depending on both neurons being close to threshold. Electrical coupling therefore controls the temporal precision and degree of both spontaneous and sensory-evoked correlated activity between interneurons, by the cooperative effects of shared synaptic depolarization and spikelet transmission.

Optogenetics applications for treating spinal cord injury.

Cases of spinal cord injury (SCI) are increasing all over the world; and in USA alone, there are 273,000 patients, which not only leads to morbidity and mortality but also results in a great economic burden. Many approaches are being used at the pre-clinical and clinical level to treat SCI including therapeutic agents, surgical decompression, stem cell therapy etc. Recently, a new approach called optogenetics has emerged in which light sensitive proteins are used to switch neurons on and off, and this approach has great potential to be used as therapy due to its specificity and rapid response in milliseconds. Few animal studies have been performed so far in which the respiratory and bladder function of rats was restored through the use of optogenetics. On the basis of promising results obtained, in the future, this approach can prove to be a valuable tool to treat patients with SCI.

Interactions between plexin-A2, plexin-A4, and semaphorin 6A control lamina-restricted projection of hippocampal mossy fibers.

Hippocampal mossy fibers project preferentially to the stratum lucidum, the proximal-most lamina of the suprapyramidal region of CA3. The molecular mechanisms that govern this lamina-restricted projection are still unknown. We examined the projection pattern of mossy fibers in mutant mice for semaphorin receptors plexin-A2 and plexin-A4, and their ligand, the transmembrane semaphorin Sema6A. We found that plexin-A2 deficiency causes a shift of mossy fibers from the suprapyramidal region to the infra- and intrapyramidal regions, while plexin-A4 deficiency induces inappropriate spreading of mossy fibers within CA3. We also report that the plexin-A2 loss-of-function phenotype is genetically suppressed by Sema6A loss of function. Based on these results, we propose a model for the lamina-restricted projection of mossy fibers: the expression of plexin-A4 on mossy fibers prevents them from entering the Sema6A-expressing suprapyramidal region of CA3 and restricts them to the proximal-most part, where Sema6A repulsive activity is attenuated by plexin-A2.

Two-photon targeted patching (TPTP) in vivo.

Two-photon-excited fluorescence laser-scanning microscopy (2PLSM) has provided a wealth of information about the spatiotemporal properties of biological processes at the single cell and population level. Because such nonlinear optical methods allow for imaging deep within biological tissue, 2PLSM can be combined with patch-clamp techniques to obtain electrophysiological recordings from specific fluorescently labeled cells in vivo. Here a protocol referred to as two-photon targeted patching (TPTP) describes a method that may be used to record from cells in the intact animal labeled by virtually any type of fluorophore. We target neurons that have been optically and genetically identified using green fluorescent protein (GFP) expressed under the control of a specific promoter. TPTP when combined with genetic approaches therefore permits electrophysiological recordings from specified neurons and their compartments, including dendrites. This technique may be repeated in the same preparation many times over the course of several hours and is equally applicable to non-neuronal cell types.

Stereotaxic gene delivery in the rodent brain.

Stereotaxic surgery has been an invaluable tool in systems neuroscience, applied in many experiments for the creation of site-targeted lesions, injection of anatomical tracers or implantation of electrodes or microdialysis probes. In this protocol, we describe stereotaxic surgery optimized for gene delivery by recombinant adeno-associated viruses and lentiviruses in mice and rats. This method allows the manipulation of gene expression in the rodent brain with excellent spatiotemporal control; essentially any brain region of choice can be targeted and cells (or a subpopulation of cells) in that region can be stably genetically altered at any postnatal developmental stage up to adulthood. Many aspects of the method, its versatility, ease of application and high reproducibility, make it an attractive approach for studying genetic, cellular and circuit functions in the brain. The entire protocol can be completed in 1-2 hours.