Nonrapid eye movement (NREM) sleep is characterized by the interaction of multiple oscillations essential for memory consolidation, alongside a dynamic arrhythmic 1/ f scale-free background that may also contribute to its functions. Recent spectral parametrization methods, such as fitting oscillation and one-and-over-F and irregular resampling auto-spectral analysis, enable the dissociation of rhythmic and arrhythmic components in the spectral domain; however, they do not resolve these processes in the time domain. Instantaneous measures of frequency, amplitude, and phase–amplitude coupling (PAC) are thus still confounded by fluctuations in arrhythmic activity. This limitation represents a pitfall for studies of NREM sleep relying on instantaneous estimates to investigate oscillatory coupling. To address this limitation, we introduce “Rhythms and Background” (RnB), a novel wavelet-based methodology designed to dynamically denoise time series data of arrhythmic interference. This enables the extraction of p...

Age-related vascular changes accompany or precede the development of Alzheimer's disease (AD) pathology. The comorbidity of AD and arterial stiffening suggests that vascular changes have a pathogenic role. Carotid artery mechanics and hemodynamics have been associated with age-related cognitive decline. However, the impact of hemodynamics and vascular mechanics on regional vulnerability within the brain has not been thoroughly explored. Compared with the arterial system, brain venous circulation in cognitive impairment is less understood despite the venous system's role in transport. To study vasculature impact on biochemistry in AD models, we must first establish the differences in vasculature mechanics and hemodynamics in a common AD model compared with healthy controls. With this baseline data, future studies on manipulating vasculature integrity in mice become feasible. Young and aged female 3xTg mice and age-matched controls were imaged using a combination of ultrasound and mass spectrometry. Wall she...

Magnetic resonance imaging (MRI) is a critical tool for translational neuroscience, but preclinical studies frequently rely on anesthesia, which alters neural activity and limits comparison with human studies. Awake rodent functional MRI (fMRI) enables investigation of brain function under physiologically relevant conditions; however, its implementation is constrained by the need for anesthesia during restraint setup. We developed and evaluated a restraint system and habituation protocol for awake rat fMRI. Ten rats were studied: an awake group and an anesthetized group (three males and two females per group). The protocol included head post implantation and an 11 d habituation period. T2-weighted anatomical and functional scans were acquired. Head motion and functional connectivity were analyzed using the RABIES pipeline and compared between groups. The modular 3D-printed restraint system developed can be assembled in under 5 min; eliminates the need for anesthesia, ear bars, and bite bars; and supports s...

Rigorous, statistically grounded experimental design is central to ethical and effective animal research. Foundational principles for statistically based Design of Experiments (DOE) were established over a century ago by Sir Ronald Fisher. They have since been augmented by modern computational tools that now enable researchers to implement designs that maximize scientific information and benefit while minimizing harms. However, many preclinical investigators are unfamiliar with formal DOE methods. Poorly designed experiments followed by inappropriate statistical analyses contribute to poor reproducibility, translational failure, and unnecessary animal use. This first paper in a three-part series introduces neuroscience researchers to the fundamentals of statistically based experimental design as a substitute for traditional two-group comparisons. Key components of a designed experiment are defined, along with the importance of correctly identifying experimental units to avoid pseudo-replication. Fisher's t...

The cerebellum is well established in subsecond motor timing, but its role in suprasecond interval timing remains unclear. Here, we investigated how cerebellar output influences time estimation over longer timescales. Male rats performed a nose-poke interval timing task in which reward availability could be predicted either from a fixed 2.5 s auditory cue (cued trials) or had to be estimated internally during uncued 3.5 s trials that demanded self-timing. Chemogenetic inhibition of the lateral cerebellar nucleus (LCN) produced bidirectional effects: delayed action initiation in predictable trials and premature (∼100–160 ms) responses when self-timing was required. Despite a slowing of movement, overall task success rates remained unchanged. Because motor slowing is likely to lead to later, not earlier, action initiation, these results implicate the LCN in computing internal time estimates. These findings demonstrate that the cerebellum integrates motor and cognitive processes for suprasecond timing, with d...

There is a critical need for robust and reliable preclinical models for posttraumatic stress disorder (PTSD) to better understand pathophysiological mechanisms and support the development of novel treatments. The single prolonged stress (SPS) model has been previously utilized to investigate various acute behavioral effects and stress hormone changes in rodents. This study paired anxiety-like and social behavioral evaluations with corticosterone assessment as a complementary physiological biomarker to determine the presence of robust and intervenable phenotypes following SPS. Sprague Dawley rats ( N  = 36, 30 male and 6 female) received SPS model induction (e.g., restraint with odorant, forced-swim, diethyl ether exposure, and isolation) or control handling. Serum corticosterone and behavioral assessments, including the open field test (OFT) and a social motivation test (SMT), were investigated at 1 and 2 weeks following SPS induction. This SPS model did not induce anxiety-like or locomotive differences as...

Social cognition, central to emotional and cognitive well-being, is particularly vulnerable to aging, where impairments can lead to isolation and functional decline. Despite compelling evidence that altered social behavior is associated with cognitive decline and dementia risk, experimental strategies for testing causative links remain scarce. To address this gap, we aimed to establish a rat model for research on social neurocognitive aging. We conducted a large-scale behavioral study in 169 male young (6 months) and aged (24–25 months) Long–Evans rats. In order to explore potential relationships among aging outcomes, we first documented individual differences in a widely validated water maze test of hippocampal learning and memory. Sociability and social novelty were then evaluated in the same subjects using the three-chamber social interaction test. Aging induced a selective shift in social novelty preference, marked by a striking familiarity bias in a substantial subpopulation of old rats, while sociabi...

Spastin is a conserved microtubule-severing enzyme mutated in hereditary spastic paraplegia. The role that spastin plays in the cell biology of axon regeneration and degeneration has recently been investigated in Drosophila . We show that the C. elegans spastin ortholog, spas-1 , is expressed in GABA motor neurons, in addition to the known expression in touch receptor neurons (TRNs) and that it is required for axon regeneration in the GABA motor neurons after in vivo laser axotomy. We identified no neuronal developmental defects in the GABA motor neurons and only minor branching variations in the TRNs. However, we show that spas-1 is required for the long-term maintenance of TRN axons in C. elegans , as older spas-1 null C. elegans show a significant increase in specific axonal morphological defects compared with the wild type as identified by confocal microscopy in aged animals. Together, our results suggest that spastin is required for regrowth and maintenance of axons in C. elegans , consistent with pre...

The progressive ratio (PR) schedule is a popular and well-established tool used to study decision-making and effort across species. In this task, subjects perform an instrumental response to receive a reinforcement, but the ratio requirement increases throughout the behavioral session. A subject's breakpoint, or the point at which the subject is no longer willing to exert the effort required to receive reinforcement, is the main behavioral readout analyzed in the PR schedule. The PR schedule has been used to investigate motivation for various reinforcers (e.g., drugs of abuse) and has been adapted to incorporate aspects of effort-related decision-making (e.g., choice between high- and low-value outcomes). However, there are notable limitations to the utility of the PR as a test of decision-making, including the confounding effects of effort and delay in that higher ratios take longer to complete, the limited behavioral response options (e.g., lever pressing), and breakpoint being a low-dimensional measurem...

Measuring neurite length is crucial in neurobiology because it provides valuable insights into the growth, development, and function of neurons. In particular, neurite length is fundamental to study neuronal development and differentiation, neurons responses to drugs, neurodegenerative diseases and neuronal plasticity. Surprisingly, there is currently a lack of tools for high throughput neurite analysis. In this article, we present CABaNe, as an open source, high throughput, rule based Image J macro for cell analysis, including their neurite length. This macro possesses a graphical interface, metadata production, as well as verification means before and after analysis. Rule based and machine learning based programming have been tested for cell identification. Cell tested were N2A, a mouse neuroblastoma cell line. After testing, we had better precision and adaptability using rule based cell identification. We challenged CABaNe with currently used techniques, which are manual or assisted. When tested on a sm...