Distinct subdivisions of subcortical U-fiber regions in the gyrencephalic ferret brain.

The human cerebrum contains a large amount of cortico-cortical association fibers. Among them, U-fibers are short-range association fibers located in white matter immediately deep to gray matter. Although U-fibers are thought to be crucial for higher cognitive functions, the organization within U-fiber regions are still unclear. Here we investigated the properties of U-fiber regions in the ferret cerebrum using neurochemical, neuronal tracing, immunohistochemical and electron microscopic techniques. We found that U-fiber regions can be subdivided into two regions, which we named outer and inner U-fiber regions. We further uncovered that outer U-fiber regions have smaller-diameter axons with thinner myelin compared with inner U-fiber regions. These findings may indicate functional complexity within U-fiber regions in the cerebrum.

Transgenic ferret models define pulmonary ionocyte diversity and function.

Speciation leads to adaptive changes in organ cellular physiology and creates challenges for studying rare cell-type functions that diverge between humans and mice. Rare cystic fibrosis transmembrane conductance regulator (CFTR)-rich pulmonary ionocytes exist throughout the cartilaginous airways of humans1,2, but limited presence and divergent biology in the proximal trachea of mice has prevented the use of traditional transgenic models to elucidate ionocyte functions in the airway. Here we describe the creation and use of conditional genetic ferret models to dissect pulmonary ionocyte biology and function by enabling ionocyte lineage tracing (FOXI1-CreERT2::ROSA-TG), ionocyte ablation (FOXI1-KO) and ionocyte-specific deletion of CFTR (FOXI1-CreERT2::CFTRL/L). By comparing these models with cystic fibrosis ferrets3,4, we demonstrate that ionocytes control airway surface liquid absorption, secretion, pH and mucus viscosity-leading to reduced airway surface liquid volume and impaired mucociliary clearance in cystic fibrosis, FOXI1-KO and FOXI1-CreERT2::CFTRL/L ferrets. These processes are regulated by CFTR-dependent ionocyte transport of Cl- and HCO3-. Single-cell transcriptomics and in vivo lineage tracing revealed three subtypes of pulmonary ionocytes and a FOXI1-lineage common rare cell progenitor for ionocytes, tuft cells and neuroendocrine cells during airway development. Thus, rare pulmonary ionocytes perform critical CFTR-dependent functions in the proximal airway that are hallmark features of cystic fibrosis airway disease. These studies provide a road map for using conditional genetics in the first non-rodent mammal to address gene function, cell biology and disease processes that have greater evolutionary conservation between humans and ferrets.

Functional ultrasound imaging reveals 3D structure of orientation domains in ferret primary visual cortex.

BACKGROUND AND PURPOSE: The sensory cortex is organized into "maps" that represent sensory space across cortical space. In primary visual cortex (V1) of highly visual mammals, multiple visual feature maps are organized into a functional architecture anchored by orientation domains: regions containing neurons preferring the same stimulus orientation. Although the pinwheel-like structure of orientation domains is well-characterized in the superficial cortical layers in dorsal regions of V1, the 3D shape of orientation domains spanning all 6 cortical layers and across dorsal and ventral regions of V1 has never been revealed. METHODS: We utilized an emerging research method in neuroscience, functional ultrasound imaging (fUS), to resolve the 3D structure of orientation domains throughout V1 in anesthetized female ferrets. fUS measures blood flow from which neuronal population activity is inferred with improved spatial resolution over fMRI. RESULTS: fUS activations in response to drifting gratings placed at multiple locations in visual space generated unique activation patterns in V1 and visual thalamus, confirming prior observations that fUS can resolve retinotopy. Iso-orientation domains, determined from clusters of activations driven by large oriented gratings, were cone-shaped and present in both dorsal and ventral regions of V1. The spacing between iso-orientation domains was consistent with spacing measured previously using optical imaging methods. CONCLUSIONS: Orientation domains are cones rather than columns. Their width and intra-domain distances may vary across dorsal and ventral regions of V1. These findings demonstrate the power of fUS at revealing 3D functional architecture in cortical regions not accessible to traditional surface imaging methods.

Reversible Inactivation of Ferret Auditory Cortex Impairs Spatial and Nonspatial Hearing.

A key question in auditory neuroscience is to what extent are brain regions functionally specialized for processing specific sound features, such as location and identity. In auditory cortex, correlations between neural activity and sounds support both the specialization of distinct cortical subfields, and encoding of multiple sound features within individual cortical areas. However, few studies have tested the contribution of auditory cortex to hearing in multiple contexts. Here we determined the role of ferret primary auditory cortex in both spatial and nonspatial hearing by reversibly inactivating the middle ectosylvian gyrus during behavior using cooling (n = 2 females) or optogenetics (n = 1 female). Optogenetic experiments used the mDLx promoter to express Channelrhodopsin-2 in GABAergic interneurons, and we confirmed both viral expression (n = 2 females) and light-driven suppression of spiking activity in auditory cortex, recorded using Neuropixels under anesthesia (n = 465 units from 2 additional untrained female ferrets). Cortical inactivation via cooling or optogenetics impaired vowel discrimination in colocated noise. Ferrets implanted with cooling loops were tested in additional conditions that revealed no deficit when identifying vowels in clean conditions, or when the temporally coincident vowel and noise were spatially separated by 180 degrees. These animals did, however, show impaired sound localization when inactivating the same auditory cortical region implicated in vowel discrimination in noise. Our results demonstrate that, as a brain region showing mixed selectivity for spatial and nonspatial features of sound, primary auditory cortex contributes to multiple forms of hearing.SIGNIFICANCE STATEMENT Neurons in primary auditory cortex are often sensitive to the location and identity of sounds. Here we inactivated auditory cortex during spatial and nonspatial listening tasks using cooling, or optogenetics. Auditory cortical inactivation impaired multiple behaviors, demonstrating a role in both the analysis of sound location and identity and confirming a functional contribution of mixed selectivity observed in neural activity. Parallel optogenetic experiments in two additional untrained ferrets linked behavior to physiology by demonstrating that expression of Channelrhodopsin-2 permitted rapid light-driven suppression of auditory cortical activity recorded under anesthesia.

Designing an assay to evaluate behavioral responses to opposite-sex conspecifics in the endangered black-footed ferret (Mustela nigripes).

The endangered black-footed ferret (ferret; Mustela nigripes) is a North American carnivore that is actively managed to reestablish self-sustaining wild populations. Behavioral abnormalities have been reported in the breeding program and may be a limiting factor for the species' success. Our goal was to design and test an assay that examines the ferret's exploratory response to odor cues in the form of soiled bedding from opposite-sex conspecifics. Across two breeding seasons, males and females were tested using a T-maze that connected their home nest box to two novel nest boxes containing two different conspecific's soiled bedding. For a control, we provided two clean bedding samples. We ran linear mixed models to determine the effect of sex, type of odor cue (soiled, clean), and order of trial (first, second) on time exploring and proportion of that time spent in each behavior. Ferrets spent the majority of time in the novel nest boxes sniffing (44%), standing alert (27%) and scratching (14%). Males explored for longer than females; however, both displayed similar behaviors. Type of cue influenced behavior, with ferrets sniffing more among soiled cues than clean cues. Habituation to the assay was also observed, with less exploration and more standing alert during the second trial of the day. This study is the first step in characterizing the ferret's exploratory response and provides information regarding vital investigatory and vigilance behaviors. The continual development of this assay to further evaluate reproductive and mate choice behaviors will facilitate more successful breeding of the species.

Sound Localization of World and Head-Centered Space in Ferrets.

The location of sounds can be described in multiple coordinate systems that are defined relative to ourselves, or the world around us. Evidence from neural recordings in animals point toward the existence of both head-centered and world-centered representations of sound location in the brain; however, it is unclear whether such neural representations have perceptual correlates in the sound localization abilities of nonhuman listeners. Here, we establish novel behavioral tests to determine the coordinate systems in which ferrets can localize sounds. We found that ferrets could learn to discriminate between sound locations that were fixed in either world-centered or head-centered space, across wide variations in sound location in the alternative coordinate system. Using probe sounds to assess broader generalization of spatial hearing, we demonstrated that in both head and world-centered tasks, animals used continuous maps of auditory space to guide behavior. Single trial responses of individual animals were sufficiently informative that we could then model sound localization using speaker position in specific coordinate systems and accurately predict ferrets' actions in held-out data. Our results demonstrate that ferrets, an animal model in which neurons are known to be tuned to sound location in egocentric and allocentric reference frames, can also localize sounds in multiple head and world-centered spaces.SIGNIFICANCE STATEMENT Humans can describe the location of sounds either relative to themselves, or in the world, independent of their momentary position. These different spaces are also represented in the activity of neurons in animals, but it is not clear whether nonhuman listeners also perceive both head and world-centered sound location. Here, we designed behavioral tasks in which ferrets discriminated between sounds using their position in the world, or relative to the head. Subjects learnt to solve both problems and generalized sound location in each space when presented with infrequent probe sounds. These findings reveal a perceptual correlate of neural sensitivity previously observed in the ferret brain and establish that, like humans, ferrets can access an auditory map of their local environment.

Tightly coupled inhibitory and excitatory functional networks in the developing primary visual cortex.

Intracortical inhibition plays a critical role in shaping activity patterns in the mature cortex. However, little is known about the structure of inhibition in early development prior to the onset of sensory experience, a time when spontaneous activity exhibits long-range correlations predictive of mature functional networks. Here, using calcium imaging of GABAergic neurons in the ferret visual cortex, we show that spontaneous activity in inhibitory neurons is already highly organized into distributed modular networks before visual experience. Inhibitory neurons exhibit spatially modular activity with long-range correlations and precise local organization that is in quantitative agreement with excitatory networks. Furthermore, excitatory and inhibitory networks are strongly co-aligned at both millimeter and cellular scales. These results demonstrate a remarkable degree of organization in inhibitory networks early in the developing cortex, providing support for computational models of self-organizing networks and suggesting a mechanism for the emergence of distributed functional networks during development.

Distinct higher-order representations of natural sounds in human and ferret auditory cortex.

Little is known about how neural representations of natural sounds differ across species. For example, speech and music play a unique role in human hearing, yet it is unclear how auditory representations of speech and music differ between humans and other animals. Using functional ultrasound imaging, we measured responses in ferrets to a set of natural and spectrotemporally matched synthetic sounds previously tested in humans. Ferrets showed similar lower-level frequency and modulation tuning to that observed in humans. But while humans showed substantially larger responses to natural vs. synthetic speech and music in non-primary regions, ferret responses to natural and synthetic sounds were closely matched throughout primary and non-primary auditory cortex, even when tested with ferret vocalizations. This finding reveals that auditory representations in humans and ferrets diverge sharply at late stages of cortical processing, potentially driven by higher-order processing demands in speech and music.

Biphasic Roles of Hedgehog Signaling in the Production and Self-Renewal of Outer Radial Glia in the Ferret Cerebral Cortex.

The neocortex, the center for higher brain function, emerged in mammals and expanded in the course of evolution. The expansion of outer radial glia (oRGs) and intermediate progenitor cells (IPCs) plays key roles in the expansion and consequential folding of the neocortex. Therefore, understanding the mechanisms of oRG and IPC expansion is important for understanding neocortical development and evolution. By using mice and human cerebral organoids, we previously revealed that hedgehog (HH) signaling expands oRGs and IPCs. Nevertheless, it remained to be determined whether HH signaling expanded oRGs and IPCs in vivo in gyrencephalic species, in which oRGs and IPCs are naturally expanded. Here, we show that HH signaling is necessary and sufficient to expand oRGs and IPCs in ferrets, a gyrencephalic species, through conserved cellular mechanisms. HH signaling increases oRG-producing division modes of ventricular radial glia (vRGs), oRG self-renewal, and IPC proliferation. Notably, HH signaling affects vRG division modes only in an early restricted phase before superficial-layer neuron production peaks. Beyond this restricted phase, HH signaling promotes oRG self-renewal. Thus, HH signaling expands oRGs and IPCs in two distinct but continuous phases during cortical development.

Biodetection of a specific odor signature in mallard feces associated with infection by low pathogenic avian influenza A virus.

Outbreaks of avian influenza virus (AIV) infection included the spread of highly pathogenic AIV in commercial poultry and backyard flocks in the spring of 2015. This resulted in estimated losses of more than $8.5 million from federal government expenditures, $1.6 billion from direct losses to produces arising from destroyed turkey and chicken egg production, and economy-wide indirect costs of $3.3 billion from impacts on retailers and the food service industries. Additionally, these outbreaks resulted in the death or depopulation of nearly 50 million domestic birds. Domesticated male ferrets (Mustela putorius furo) were trained to display a specific conditioned behavior (i.e. active scratch alert) in response to feces from AIV-infected mallards in comparison to feces from healthy ducks. In order to establish that ferrets were identifying samples based on odors associated with infection, additional experiments controlled for potentially confounding effects, such as: individual duck identity, housing and feed, inoculation concentration, and day of sample collection (post-infection). A final experiment revealed that trained ferrets could detect AIV infection status even in the presence of samples from mallards inoculated with Newcastle disease virus or infectious laryngotracheitis virus. These results indicate that mammalian biodetectors are capable of discriminating the specific odors emitted from the feces of non-infected versus AIV infected mallards, suggesting that the health status of waterfowl can be evaluated non-invasively for AIV infection via monitoring of volatile fecal metabolites. Furthermore, in situ monitoring using trained biodetectors may be an effective tool for assessing population health.

Development of visual motion integration involves coordination of multiple cortical stages.

A central feature of cortical function is hierarchical processing of information. Little is currently known about how cortical processing cascades develop. Here, we investigate the joint development of two nodes of the ferret's visual motion pathway, primary visual cortex (V1), and higher-level area PSS. In adult animals, motion processing transitions from local to global computations between these areas. We now show that PSS global motion signals emerge a week after the development of V1 and PSS direction selectivity. Crucially, V1 responses to more complex motion stimuli change in parallel, in a manner consistent with supporting increased PSS motion integration. At the same time, these V1 responses depend on feedback from PSS. Our findings suggest that development does not just proceed in parallel in different visual areas, it is coordinated across network nodes. This has important implications for understanding how visual experience and developmental disorders can influence the developing visual system.

Dissociation of task engagement and arousal effects in auditory cortex and midbrain.

Both generalized arousal and engagement in a specific task influence sensory neural processing. To isolate effects of these state variables in the auditory system, we recorded single-unit activity from primary auditory cortex (A1) and inferior colliculus (IC) of ferrets during a tone detection task, while monitoring arousal via changes in pupil size. We used a generalized linear model to assess the influence of task engagement and pupil size on sound-evoked activity. In both areas, these two variables affected independent neural populations. Pupil size effects were more prominent in IC, while pupil and task engagement effects were equally likely in A1. Task engagement was correlated with larger pupil; thus, some apparent effects of task engagement should in fact be attributed to fluctuations in pupil size. These results indicate a hierarchy of auditory processing, where generalized arousal enhances activity in midbrain, and effects specific to task engagement become more prominent in cortex.

Cryopreservation of ferret (Mustela putorius furo) sperm collected by rectal massage and electroejaculation: Comparison of a decelerating and an accelerating freezing rate protocol.

The domestic ferret (Mustela putorius furo) provides a good model for developing new reproductive technologies for use with threatened related species. Such technologies could also be used in the reproductive management of this pet species. The present work reports an improved freezing protocol for ferret sperm. Semen was collected by electroejaculation plus rectal massage (in an attempt to reduce the electrical stimulation necessary) from five adult male ferrets, and then subjected to one of two freezing protocols: (a) from 5 to -35°C at 40°C/min, then from -35 to -65°C at 17°C/min, and finally from -65 to -85°C at 3°C/min-a decelerating freezing rate; and (b) from 5 to - 10°C at 5°C/min, and then from -10 to -130°C at 60°C/min-an accelerating freezing rate. After thawing, the viability and acrosomal integrity of the sperm frozen via the two-step accelerating method were better than those frozen via the three-step decelerating method (43.3 ± 3.5% and 71.2 ± 3.4% compared with 29.7 ± 3.7% and 58.8 ± 3.4% respectively; p < .05). No differences were seen between the methods with respect to sperm motility variables; most sperm (>90%) remained static with both freezing methods. In conclusion, although the method with accelerating freezing rate was associated with better post-thaw sperm viability and acrosome integrity values, neither of the two freezing methods tested provided adequate motility results after thawing. Combining rectal massage with electrical stimuli seemed to reduce the number of the latter required for successful sperm collection.

¿Puedo diagnosticar la hipoglucemia en hurones domésticos (Mustela putorius furo) con el uso de glucómetros portátiles? / Can I diagnose hypoglycemia in domestic ferrets (Mustela putorius furo) with the use of portable glucometers?

OBJETIVO: El diagnóstico de la hipoglucemia en hurones (Mustela putorius furo) se basa en la detección de un nivel de glucosa en la sangre inferior a 60- 70 mg / dl. Una concentración de glucosa en sangre por debajo de estos niveles es muy sugestiva de tumor de células beta pancreáticas (insulinoma), que comprende aproximadamente el 25% de todas las neoplasias diagnosticadas en esta especie. En este estudio se pretende comprobar si los bajos valores de glucosa en sangre obtenidos mediante el uso de glucómetros portátiles son fiables como método de diagnóstico de hipoglucemia en esta especie. MATERIAL Y MÉTODOS: En este estudio se analizan 31 muestras de sangre de hurones de distinta edad y sexo, mediante el uso de tres tipos de glucómetros portátiles, comparando dichos resultados con las medidas obtenidas mediante un analizador de bioquímica líquida que usa el método de la glucosa oxidasa, considerado como uno de los métodos estándar de medida. RESULTADOS: Uno de los tres glucómetros utilizado en este estudio subestima los valores de glucosa en sangre en esta especie, mientras que otro los sobreestima, lo que supone un error sistemático (predecible) de los equipos que se podría solucionar calibrándolos para esta especie. El otro glucómetro que presenta mayor concordancia con los valores de referencia presenta un error aleatorio elevado (impredecible) más difícil de corregir. Por tanto ninguno de los tres equipos sería adecuado para realizar un diagnóstico fiable de hipoglucemia en hurones. CONCLUSIONES: Estos resultados indican que las concentraciones de glucosa en sangre en hurones deben confirmarse con un analizador de laboratorio validado para realizar el diagnóstico de hipoglucemia de forma fiable, y los glucómetros usarse sólo para hacer un seguimiento de la enfermedad

AIMS: The diagnosis of hypoglycemia in ferrets (Mustela putorius furo) is based on the detection of a blood glucose level below 60-70 mg / dl. A blood glucose concentration below these levels is very suggestive of pancreatic beta cell tumor (insulinoma), which comprises of approximately 25% of all neoplasms diagnosed in this species. This study aims to check if the low blood glucose values obtained through the use of portable glucometers are reliable as a method of diagnosing hypoglycemia in this species. MATERIAL AND METHODS: In this study, 31 samples of ferrets blood of different ages and sexes are analyzed, using three types of portable glucometers, comparing these results with the measurements obtained by a liquid biochemistry analyzer using the glucose oxidase method, considered as one of the standard measurement methods. RESULTS: One of the three glucometers used in this study underestimates blood glucose values in this species, while another overestimates them, which is a systematic error (predictable) of the equipment that could be solved by calibrating them for this species. The other glucometer is more consistent with the reference values, but has a high random error (unpredictable), that is more difficult to correct. Therefore none of the three teams would be adequate to make a reliable diagnosis of hypoglycemia in ferrets. CONCLUSIONS: These results indicate that blood glucose concentrations in ferrets should be confirmed with a validated laboratory analyzer to reliably diagnose hypoglycemia, and blood glucose meters used only to monitor the disease

Influence of vitamin E and carcass feeding supplementation on fecal glucocorticoid and androgen metabolites in male black-footed ferrets (Mustela nigripes).

In recent years, the ex situ population of the endangered black-footed ferret (Mustela nigripes; ferret) has experienced a decline in normal sperm morphology (from 50% to 20%), which may be linked to inbreeding depression and/or a dietary change. We examined the effects of adding carcass and vitamin E to the diet on stress and reproductive biomarkers in male ferrets (n = 42 males including 16 juveniles and 26 adults) housed at the U.S. Fish and Wildlife National Black-footed Ferret Conservation Center (Carr, CO, USA). Fecal samples (3x/week) were collected from November and December (pre-breeding season, no diet change), February through May (breeding season, diet change) and June (post-breeding season, diet change) and analyzed for fecal glucocorticoid metabolites (FGM) via a corticosterone enzyme immunoassay (EIA). A subset of samples from adult males (n = 15) were analyzed for fecal androgen metabolites (FAM) via a testosterone EIA. We first used a linear mixed effects model to identify the important fixed effects among meat treatment, vitamin E treatment, age class (juvenile or adult), and all possible interactions on each hormone. We then examined the important factor's effects across seasons using the non-parametric Friedman test. We found that age did not influence (p = 0.33) FGMs; however there was a significant effect of meat treatment on FGM (p = 0.04) and an effect of vitamin E on FAMs (p<0.10). When fed carcass, FGMs declined (p<0.001) from pre- to the during the breeding season time period, but was similar (p>0.05) between during and post-breeding season periods. Males that were not fed carcass had higher (p<0.05) FGMs during the breeding season compared to pre- and post-breeding season and FGMs were lower (p<0.05) in the post-breeding season compared to pre-breeding season. Males fed with carcass had lower (p<0.001) FGM than males that were not fed carcass during both the pre-breeding and the breeding season but not during the post-breeding season (p>0.05). Males supplemented with vitamin E had higher (p<0.001) FAM than non-supplemented males during the breeding season only. For both groups, FAM was highest (p<0.05) during the breeding season. In conclusion, adding carcass to the diet can reduce glucocorticoid production and adding vitamin E can increase testosterone during the breeding season, which may influence reproductive success.

Seasonal weight changes in laboratory ferrets.

Ferrets (Mustela putorius furo) are a valuable animal model used in biomedical research. Like many animals, ferrets undergo significant variation in body weight seasonally, affected by photoperiod, and these variations complicate the use weight as an indicator of health status. To overcome this requires a better understanding of these seasonal weight changes. We provide a normative weight data set for the female ferret accounting for seasonal changes, and also investigate the effect of fluid regulation on weight change. Female ferrets (n = 39) underwent behavioural testing from May 2017 to August 2019 and were weighed daily, while housed in an animal care facility with controlled light exposure. In the winter (October to March), animals experienced 10 hours of light and 14 hours of dark, while in summer (March to October), this contingency was reversed. Individual animals varied in their body weight from approximately 700 to 1200 g. However, weights fluctuated with light cycle, with animals losing weight in summer, and gaining weight in winter such that they fluctuated between approximately 80% and 120% of their long-term average. Ferrets were weighed as part of their health assessment while experiencing water regulation for behavioural training. Water regulation superimposed additional weight changes on these seasonal fluctuations, with weight loss during the 5-day water regulation period being greater in summer than winter. Analysing the data with a Generalised Linear Model confirmed that the percentage decrease in weight per week was relatively constant throughout the summer months, while the percentage increase in body weight per week in winter decreased through the season. Finally, we noted that the timing of oestrus was reliably triggered by the increase in day length in spring. These data establish a normative benchmark for seasonal weight variation in female ferrets that can be incorporated into the health assessment of an animal's condition.

Synaptic and intrinsic mechanisms underlying development of cortical direction selectivity.

Modifications of synaptic inputs and cell-intrinsic properties both contribute to neuronal plasticity and development. To better understand these mechanisms, we undertook an intracellular analysis of the development of direction selectivity in the ferret visual cortex, which occurs rapidly over a few days after eye opening. We found strong evidence of developmental changes in linear spatiotemporal receptive fields of simple cells, implying alterations in circuit inputs. Further, this receptive field plasticity was accompanied by increases in near-spike-threshold excitability and input-output gain that resulted in dramatically increased spiking responses in the experienced state. Increases in subthreshold membrane responses induced by the receptive field plasticity and the increased input-output spiking gain were both necessary to explain the elevated firing rates in experienced ferrets. These results demonstrate that cortical direction selectivity develops through a combination of plasticity in inputs and in cell-intrinsic properties.

Geriatric Ferrets.

Ferrets are considered geriatric at 3 years of age. Twice-yearly physical examinations with additional testing finds problems and leads to prompt treatments that extend quality of life for the ferret. Bloodwork, urinalysis, dental examination, and imaging including radiographs and ultrasonography should be performed at least annually. Specialized testing includes echocardiography, electrocardiology, blood pressure, computed tomography scanning, and dental radiography depending on the ferret. Common problems include dental disease, cardiomyopathy, renal disease, musculoskeletal disorders, chronic gastrointestinal disorders, and neoplasias. Insulinoma, adrenal neoplasia, lymphoma, and skin tumors are commonly found neoplasias. Medical and surgical treatments and adjunctive complimentary therapies are discussed.

Imperfect transparency and camouflage in glass frogs.

Camouflage patterns prevent detection and/or recognition by matching the background, disrupting edges, or mimicking particular background features. In variable habitats, however, a single pattern cannot match all available sites all of the time, and efficacy may therefore be reduced. Active color change provides an alternative where coloration can be altered to match local conditions, but again efficacy may be limited by the speed of change and range of patterns available. Transparency, on the other hand, creates high-fidelity camouflage that changes instantaneously to match any substrate but is potentially compromised in terrestrial environments where image distortion may be more obvious than in water. Glass frogs are one example of terrestrial transparency and are well known for their transparent ventral skin through which their bones, intestines, and beating hearts can be seen. However, sparse dorsal pigmentation means that these frogs are better described as translucent. To investigate whether this imperfect transparency acts as camouflage, we used in situ behavioral trials, visual modeling, and laboratory psychophysics. We found that the perceived luminance of the frogs changed depending on the immediate background, lowering detectability and increasing survival when compared to opaque frogs. Moreover, this change was greatest for the legs, which surround the body at rest and create a diffuse transition from background to frog luminance rather than a sharp, highly salient edge. This passive change in luminance, without significant modification of hue, suggests a camouflage strategy, "edge diffusion," distinct from both transparency and active color change.

Experience-Dependent Reorganization Drives Development of a Binocularly Unified Cortical Representation of Orientation.

Across sensory areas, neural microcircuits consolidate streams of information into unified representations of the external world. In the carnivore visual cortex, where eye-specific inputs first converge, it has been posited that a single, binocularly aligned modular orientation representation develops independent of sensory experience. In this study of ferret visual cortex using in vivo calcium imaging, we find evidence for a different developmental process. Early in development, contralateral, ipsilateral, or binocular stimulation each yield well-organized modular representations of orientation that display features of mature cortex. However, comparison of these representations reveals considerable misalignment that is evident at both modular and cellular scales. Experience-dependent processes drive reorganization of these three representations toward a single binocularly aligned representation resembling the early binocular representation through shifts in cellular orientation preference. Thus, while orderly modular networks of orientation preference initially arise independent of visual experience, experience is critical for the alignment of these early representations.