Pigmented Ferrets Research Articles

Auditory gap-in-noise detection behavior in ferrets and humans.

The precise encoding of temporal features of auditory stimuli by the mammalian auditory system is critical to the perception of biologically important sounds, including vocalizations, speech, and music. In this study, auditory gap-detection behavior was evaluated in adult pigmented ferrets (Mustelid putorius furo) using bandpassed stimuli designed to widely sample the ferret’s behavioral and physiological audiogram. Animals were tested under positive operant conditioning, with psychometric functions constructed in response to gap-in-noise lengths ranging from 3 to 270 ms. Using a modified version of this gap-detection task, with the same stimulus frequency parameters, we also tested a cohort of normal-hearing human subjects. Gap-detection thresholds were computed from psychometric curves transformed according to signal detection theory, revealing that for both ferrets and humans, detection sensitivity was worse for silent gaps embedded within low-frequency noise compared with high-frequency or broadband stimuli. Additional psychometric function analysis of ferret behavior indicated effects of stimulus spectral content on aspects of behavioral performance related to decision-making processes, with animals displaying improved sensitivity for broadband gap-in-noise detection. Reaction times derived from unconditioned head-orienting data and the time from stimulus onset to reward spout activation varied with the stimulus frequency content and gap length, as well as the approach-to-target choice and reward location. The present study represents a comprehensive evaluation of gap-detection behavior in ferrets, while similarities in performance with our human subjects confirm the use of the ferret as an appropriate model of temporal processing.

Plasticity of spatial hearing: behavioural effects of cortical inactivation

The contribution of auditory cortex to spatial information processing was explored behaviourally in adult ferrets by reversibly deactivating different cortical areas by subdural placement of a polymer that released the GABAA agonist muscimol over a period of weeks. The spatial extent and time course of cortical inactivation were determined electrophysiologically. Muscimol-Elvax was placed bilaterally over the anterior (AEG), middle (MEG) or posterior ectosylvian gyrus (PEG), so that different regions of the auditory cortex could be deactivated in different cases. Sound localization accuracy in the horizontal plane was assessed by measuring both the initial head orienting and approach-to-target responses made by the animals. Head orienting behaviour was unaffected by silencing any region of the auditory cortex, whereas the accuracy of approach-to-target responses to brief sounds (40 ms noise bursts) was reduced by muscimol-Elvax but not by drug-free implants. Modest but significant localization impairments were observed after deactivating the MEG, AEG or PEG, although the largest deficits were produced in animals in which the MEG, where the primary auditory fields are located, was silenced. We also examined experience-induced spatial plasticity by reversibly plugging one ear. In control animals, localization accuracy for both approach-to-target and head orienting responses was initially impaired by monaural occlusion, but recovered with training over the next few days. Deactivating any part of the auditory cortex resulted in less complete recovery than in controls, with the largest deficits observed after silencing the higher-level cortical areas in the AEG and PEG. Although suggesting that each region of auditory cortex contributes to spatial learning, differences in the localization deficits and degree of adaptation between groups imply a regional specialization in the processing of spatial information across the auditory cortex.

Sensitivity to relative disparity in early visual cortex of pigmented and albino ferrets

To investigate binocular interactions as the neuronal substrate for disparity sensitivity in the ferret (Mustela putorius furo), we measured the effects of relative horizontal disparities on responses of neurons in areas 17 and 18 of the visual cortex. Stimulation by moving bars and sinusoidal gratings showed that about half of our sample in pigmented ferrets was sensitive to relative horizontal disparity. This also included many neurons, which were classified as only monocularly activated when testing either eye alone. However, the tuning width was about two or three times coarser (median tuning width 4 degrees of visual angle) than that in the cat. In albino ferrets, only 8% of the neurons in the early visual cortex displayed some sort of disparity-dependent binocular interactions, but none could be clearly identified as relative disparity-coding neuron.

Relay of visual information to the lateral geniculate nucleus and the visual cortex in albino ferrets.

The abnormal organization of the central visual pathways in the albino ferret has been characterized anatomically and physiologically. Recordings in dorsal lateral geniculate nucleus of the albino ferret show that lamina A1, which receives an aberrant projection from the contralateral eye, contains an extensive representation of the ipsilateral visual hemifield with receptive fields located up to 35 degrees from the vertical meridian. This is not the case in pigmented ferrets, for which the vast majority of units, activated through either the contralateral or ipsilateral eye, have receptive fields confined to the contralateral hemifield. The few fields found in the ipsilateral hemifield are driven through the contralateral eye and none is more than 10 degrees from the midline. Cortical topography was studied by making closely spaced electrode penetrations across the area 17/18 border. In pigmented animals, the reversal of topography at the border is characterized by units with receptive fields centered a few degrees into the ipsilateral hemifield. In 22 of 25 albinos, the "Boston" aberrant topography was found: the representation of the vertical meridian is within area 17, rather than at the area 17/18 border. Instead, at the area 17/18 border, there is a reversal in the topographic progression at up to 30 degrees into the ipsilateral hemifield. This pattern was most pronounced in the upper visual field. In agreement with the "Boston" physiology, injections of retrograde tracer made in area 17 usually label neurons in either lamina A or the part of lamina A1 that is aberrantly innervated by the contralateral eye. A column of labeled cells extending through all geniculate layers is rarely seen in albinos, although this is commonly the pattern in pigmented ferrets.

Visual field defects in albino ferrets ( Mustela putorius furo)

The extent of the horizontal visual field was determined behaviourally in 4 pigmented and 5 albino ferrets ( Mustela putorius furo, Carnivora, Mammalia) using perimetry. During binocular vision, all pigmented and three albino ferrets responded equally well to stimuli presented anywhere along the horizontal perimeter in the central 180° of the visual field. The remaining two albinos had a visual field defect in the right hemifield (>30° eccentricity). During monocular vision, a significant difference between the visual fields of pigmented and albino ferrets became apparent. In pigmented ferrets, the visual field of each eye included the ipsilateral (temporal) and a substantial portion of the contralateral (nasal) hemifield. In albinos, the visual field of each eye was limited to the ipsilateral hemifield and reactions to visual stimuli abruptly declined directly beyond the vertical meridian.

Morphological changes in the neuronal substrate for the optokinetic reflex in albino ferrets.

Albino mammals show very characteristic deficits in their optokinetic system, and albino ferrets are even optokinetically blind. To investigate the neuronal causes for this defect we compared the morphology of retinal slip cells in the pretectal nucleus of the optic tract and the dorsal terminal nucleus of the accessory optic system (NOT-DTN) in pigmented and albino ferrets (Mustela putorius furo) using retrograde tracing techniques. After tracer injections into the inferior olive, equal numbers of NOT-DTN neurons were retrogradely labelled in pigmented and albino animals. However, NOT-DTN cells in albino ferrets had fewer stem dendrites, and the cumulative dendritic length was reduced by 30% when compared with NOT-DTN neurons in pigmented animals. In addition, the prominent network formed by distal dendrites observed in the NOT-DTN of pigmented ferrets was largely diminished in albinos. Taken together with behavioural and physiological data, these findings indicate that the NOT-DTN as the main visuomotor interface in the optokinetic system is clearly defective in albino ferrets.

The effects of monocular enucleation on ganglion cell number and terminal distribution in the ferret's retinal pathway.

Anterograde and retrograde tracing techniques were used to examine the effects of removing one eye at birth on the remaining uncrossed retinal pathway in adult ferrets. After enucleation, the adult number of labelled ganglion cells projecting ipsilaterally changed from an average of 6068 in normal pigmented ferrets to an average of 7813 (29% increase) in pigmented enucleates. The change in albino ferrets was from 1455 in normals to 2319 in enucleates (59% increase). Labelled cells scattered across nasal retina accounted for over half the increase in the uncrossed population. After neonatal enucleation, the volume of lateral geniculate nucleus occupied by the uncrossed projection increased substantially: five-fold in pigmented animals and 20-fold in albinos. These results suggest that neonatal removal of one eye has a greater effect on the distribution of uncrossed terminals than on the survival of uncrossed ganglion cells. There was also an increase in the total number of axons in the surviving optic nerve of both pigmented and albino ferrets (93,000 in enucleates compared with 79,000 in normal animals), which cannot be simply explained as a disruption of binocular competition.

The development of retinal ganglion cell decussation patterns in postnatal pigmented and albino ferrets.

The decussation patterns of retinal ganglion cells in postnatal pigmented and albino ferrets were examined by using retrograde axonal tracers. Following unilateral injections into the optic pathway of newborn pigmented ferrets, approximately 13,000 cells were labelled in the ipsilateral retina. The majority (11,500) of these were located in temporal retina. Postnatally, the numbers of cells projecting ipsilaterally from temporal retina fell by 49%. High rates of loss were observed in both the smaller uncrossed projection from nasal retina (92%) and also in the crossed projection from temporal retina (84%). After injections on the day of birth, a decussation line was not obvious in the crossed projection: > or = 14,000 labelled cells were found in temporal retina. Double tracer studies showed that very few of these cells had axons which projected bilaterally. The numbers of ipsilaterally projecting cells labelled in neonatal albino ferrets was dramatically reduced. Only approximately 2500 were labelled in temporal retina following injections at birth. As in pigmented ferrets, about half of these cells subsequently died. The reduced uncrossed projection in albino neonates was associated with an increase in the crossed projection from temporal retina, in which approximately 21,000 cells were labelled following injections at birth. These results suggest that differential postnatal ganglion cell death establishes the adult decussation pattern in the contralateral retinal projection but merely refines the pattern already established in the uncrossed projection. Postnatal ganglion cell death plays no significant role in generating the abnormal projections found in albino ferrets.

Dendritic development in the dorsal lateral geniculate nucleus of ferrets in the postnatal absence of retinal input: A golgi study

In order to determine the ongoing role of retinal fibers in the development of dorsal lateral geniculate nucleus (dLGN) neurons during postnatal development, the development of dLGN neurons in the postnatal absence of retinal input was studied in pigmented ferrets using the Golgi-Hortega technique. The development of four dLGN cell classes, defined on the basis of somatic and dendritic morphology, was described previously in normal ferrets (Sutton and Brunso-Bechtold, 1991, J. Comp. Neurol. 309:71-85). The present results indicate that the morphological development of dLGN neurons is strikingly similar in normal and experimental ferrets. The exuberant dendritic appendages that appear after eye opening in normal ferrets are overproduced and eliminated in the postnatal absence of retinal input; however, the final reduction of these transient appendages is delayed. Because exuberant appendages develop in the absence of retinal input, their production cannot depend upon visual experience. Differences in cell body size between normal and experimental ferrets are apparent only after neurons can be classified at the end of the first postnatal month. Cell body size is markedly reduced for class 1 neurons; class 2 cells also are reduced in size but to a far lesser extent. As there is a general trend for class 1 neurons to have the functional properties of Y-cells, it is likely that the dLGN neurons most affected by the absence of retinal input also are Y-cells.

The segregation of ON- and OFF-center responses in the lateral geniculate nucleus of normal and monocularly enucleated ferrets.

We have investigated the distribution of ON- and OFF-center responses in the lateral geniculate nucleus of ferrets with normal and abnormal retinal projections. Electrophysiological recordings in normal pigmented animals confirm previous studies on mustelids showing that ON-center responses are found in the anterior, inner parts of laminae A and A1 and OFF-center responses in posterior, outer leaflets. In albino animals, lamina A displays normal patterns of ON/OFF segregation but in lamina A1', which receives an abnormal crossed retinal projection, no consistent patterns of segregation are found. Following monocular enucleation on the day of birth, the uncrossed projection in pigmented ferrets remains expanded across the LGN. Anatomically and physiologically, this projection is segregated into two leaflets: an anterior, inner ON-center leaflet and a posterior outer OFF-center leaflet. We conclude that the persistence of ON/OFF segregation, independent of geniculate location, suggests that self-sorting of retinal input is an important factor in generating the segregation.

A golgi study of dendritic development in the dorsal lateral geniculate nucleus of normal ferrets

The development of neurons in the dorsal lateral geniculate nucleus (dLGN) of pigmented ferrets was studied by using the Golgi-Hortega technique. In adult ferrets, four dLGN cell classes were defined on the basis of somatic and dendritic morphology. Classes 1 and 2 were divided into stellate and oriented subtypes. Class 1 and 4 cells are characterized by filiform appendages, class 2 cells by club-like appendages, and class 3 cells by stalked appendages. At birth, dLGN neurons have simple dendritic arbors. During the first postnatal week, dendritic length and proximal branching density increase markedly. By postnatal day 21 (P21), dendritic morphology begins to take on mature characteristics and by the time of eye opening (P30-P35), most neurons can be classified. Also by that time, dLGN cells are covered with abundant filiform appendages. Developmental changes in appendage density were quantified for class 1 stellate cells. These data reveal that appendage density reaches a peak at P56, decreases sharply until P90, and then gradually declines to mature levels by P180. Elaboration and elimination of transient appendages occurs centrifugally; at maturity appendage density remains greater distally.

Albino gene dosage and retinal decussation patterns in the pigmented ferrret

We have examined the retinal decussation patterns in pigmented ferrets that were either wild-type sable or heterozygous with one albino gene. Unilateral injections of horseradish peroxidase were made into the optic tract and labeled ganglion cells visualized in retinal wholemounts. In both wild-type and heterozygous ferrets, those ganglion cells in the temporal retina with the largest cell bodies projected only to the contralateral side of the brain. The total number of ipsilaterally projecting ganglion cells did not differ with the genotype of the animal. The numbers ranged from 5471-6759 cells. Unlike the cat, there is no difference in retinal decussation patterns in wild-type sable ferrets and heterozygous ferrets carrying one albino gene.

Evidence for the delayed expression of a brainstem abnormality in albino ferrets.

Previous reports have suggested that neurons of the medial superior olivary nucleus in albino cats and rabbits are smaller than those in normally pigmented strains. In this investigation, the mean cross-sectional areas of neuronal perikarya in the medial superior olivary nucleus of pigmented and albino ferrets were compared at juvenile (14 weeks) and adult (greater than six months) ages. The mean cross-sectional area of the perikarya in adult albino ferrets was found to be 45% smaller than that of adult pigmented ferrets, confirming observations in cat and rabbit. The same comparison in juvenile ferrets, however, showed no significant differences. These results suggest that the mechanisms producing the abnormality of the auditory pathways differ from those responsible for the production of abnormal retinofugal pathways in albinos.

Spatial properties of neurones in the lateral geniculate nucleus of the pigmented ferret.

We used quantitative electrophysiological techniques to study the spatial properties of single units recorded extracellularly in the lateral geniculate and perigeniculate nuclei of the adult pigmented ferret. All neurones examined had approximately circular receptive fields, whose central regions gave responses antagonistic to those elicited from the surrounds. We presented sinusoidally modulated grating patterns, either drifting or counterphased, to obtain spatial frequency tuning curves, contrast sensitivity functions and to assess linearity or non-linearity of each neurone's response. In the ferret, as in other species, two types of lateral geniculate neurone could be distinguished, and we have termed these X-cells and Y-cells; both groups responded briskly to visual stimulation but X-cells gave sustained and linear responses whereas Y-cells responded transiently and non-linearly. Perigeniculate cells gave linear responses. For neurones in the lateral geniculate and perigeniculate nuclei, both the limit of spatial resolution ('acuity') and optimum spatial frequency were inversely related to receptive field eccentricity and the diameter of the receptive field centre. We recorded geniculate neurones in the ferret with acuities up to 3 cycles deg-1 and contrast sensitivities up to 114, values that are lower than those found previously for many geniculate cells in the cat.

Retinal decussation patterns in pigmented and albino ferrets

The decussation patterns of retinal ganglion cells in adult pigmented and albino ferrets were determined from the distribution of cells labelled after large unilateral injections of horseradish peroxidase into the visual pathway, involving the lateral geniculate nucleus and fibres of passage to the superior colliculus. About 6000 retinal ganglion cells project ipsilaterally in pigmented ferrets compared with only about 1500 in albino ferrets. In both strains, the vast majority of these cells (99 and 87% in pigmented and albino animals, respectively) are located in the temporal crescent, although we describe one albino ferret in which an aberrant uncrossed projection arises from nasal retina. In pigmented ferrets, there is a sharp nasotemporal division that runs through the area centralis; a small proportion of the ganglion cells in temporal crescent (< 10%) does project contralaterally. In albinos, however, the majority of cells in temporal retina project contralaterally. There is no clear nasotemporal division in the albino retina; the density of uncrossed ganglion cells is reduced throughout temporal crescent and at no location exceeds the comparable density of the crossed projection. The peak density within the reduced uncrossed projection is also displaced away from the area centralis into temporal retina. Analysis of cell type on the basis of soma size indicates that whereas large horseradish peroxidase injections into the visual pathway of pigmented ferrets label all types of ganglion cell in the crossed projection, injections restricted to the superior colliculus label only those ganglion cells with large or small somata. The distribution of cell sizes in the crossed projection from temporal retina is biased towards small cells in the pigmented ferret but in albinos resembles that seen in the crossed projection from nasal retina. Thus the adult pigmented ferret has both a well developed nasotemporal division in which decussation lines are obvious in the crossed and uncrossed pathways and also, unlike rodents but like cats, a class of ganglion cell that does not project to the superior colliculus. The albino mutation both reduces the uncrossed projection throughout temporal retina, although the reduction is greatest close to the area centralis and also commensurately increases the crossed projection from temporal retina.

Cholinergic innervation of ferret visual system

The distribution of acetylcholinesterase and choline acetyltransferase in primary visual areas of adult pigmented ferret was determined with cholinesterase histochemistry and choline acetyltransferase immunohistochemistry. In all visual areas the distribution of acetylcholinesterase in the neuropil closely matches that of choline acetyltransferase. In the cerebral cortex acetylcholinesterase and choline acetyltransferase are associated with axons found in every cortical layer and in the white matter. Area 17, identified by Nissl architectonics and cytochrome oxidase histochemistry, is distinguished by having a relatively low density of choline acetyltransferase- and acetylcholinesterase-stained axons in layer IV. Certain cortical non-pyramidal cell types show moderate staining for acetylcholinesterase after relatively long incubations, but no choline acetyltransferase-positive cells are observed in the cortex. In the lateral geniculate nucleus and superior colliculus the levels of choline acetyltransferase and acetylcholinesterase are considerably higher than in cerebral cortex and choline acetyltransferase-stained axons there display prominent varicosities. The distribution of choline acetyltransferase and acetylcholinesterase in the neuropil of lateral geniculate nucleus and superior colliculus of ferret shows marked laminar variation. For instance, in the lateral geniculate nucleus, the levels of acetylcholinesterase and choline acetyltransferase in the “On” sublaminae of laminae A and A1 are higher than the “Off” sublaminae. In the superficial layers of the superior colliculus the levels of choline acetyltransferase and acetylcholinesterase are highest in the stratum zonale and lowest in the stratum opticum; in the intermediate gray layer of the superior colliculus acetylcholinesterase- and choline acetyltransferase-stained fibres are distributed into dense patches. As in cortex, choline acetyltransferase-positive cell bodies are not found in the lateral geniculate nucleus or superior colliculus and acetylcholinesterase-stained cell bodies are visible only after long incubations. Cell bodies staining positively for choline acetyltransferase are found in a satellite of the superior colliculus, the parabigeminal nucleus.

Distribution of ganglion cells in the retina of adult pigmented ferret

The retinal ganglion cell distribution in adult pigmented ferret was mapped in Nissl-stained retinas and in retinas back-filled with HRP after large bilateral injections of the enzyme into the brain. In common with other carnivores the ferret has an area of peak cell density equivalent to the area centralis and a prominent visual streak of high cell density extending horizontally across the retina. The maximum ganglion cell densities for the retinas were estimated to be 3500–5200 cells/mm 2 in Nissl-stained, dehydrated retinas and 3300–4300 cells/mm 2 in HRP-labelled, undehydrated retinas. Three cell types were distinguished in the HRP-labelled retinas and they appear to correspond to alpha-, beta- and gamma-cell types of cat retina. However, unlike in cat, the retinal ganglion cells of the ferret do not consistently fall into 3 distinct groups with respect to cell size, nor is there a tendency for the cells in the area centralis to be smaller than those in the peripheral retina. Estimates for the total number of ganglion cells of 82,000 and 88,000 were obtained from Nissl-stained retinas, and of 74,000, 75,000 and 78,000 from HRP-labelled retinas.