When enough is enough: Optimising monitoring effort for large-scale wolf population size estimation in the Italian Alps.

The ongoing expansion of wolf (Canis lupus) populations in Europe has led to a growing demand for up-to-date abundance estimates. Non-invasive genetic sampling (NGS) is now widely used to monitor wolves, as it allows individual identification and abundance estimation without physically capturing individuals. However, NGS is resource-intensive, partly due to the elusive behaviour and wide distribution of wolves, as well as the cost of DNA analyses. Optimisation of sampling strategies is therefore a requirement for the long-term sustainability of wolf monitoring programs. Using data from the 2020-2021 Italian Alpine wolf monitoring, we investigate how (i) reducing the number of samples genotyped, (ii) reducing the number of transects, and (iii) reducing the number of repetitions of each search transect impacted spatial capture-recapture population size estimates. Our study revealed that a 25% reduction in the number of transects or, alternatively, a 50% reduction in the maximum number of repetitions yielded abundance estimates comparable to those obtained using the entire dataset. These modifications would result in a 2046 km reduction in total transect length and 19,628 km reduction in total distance searched. Further reducing the number of transects resulted in up to 15% lower and up to 17% less precise abundance estimates. Reducing only the number of genotyped samples led to higher (5%) and less precise (20%) abundance estimates. Randomly subsampling genotyped samples reduced the number of detections per individual, whereas subsampling search transects resulted in a less pronounced decrease in both the total number of detections and individuals detected. Our work shows how it is possible to optimise wolf monitoring by reducing search effort while maintaining the quality of abundance estimates, by adopting a modelling framework that uses a first survey dataset. We further provide general guidelines on how to optimise sampling effort when using spatial capture-recapture in large-scale monitoring programmes.

Functional selectivity of interhemispheric connections in cat visual cortex.

The functional specificity of callosal connections was investigated in visual areas 17 and 18 of adult cats, by combining in vivo optical imaging of intrinsic signals with labeling of callosal axons. Local injections of neuronal tracers were performed in one hemisphere and eight single callosal axons were reconstructed in the opposite hemisphere. The distributions of injection sites and callosal axon terminals were analyzed with respect to functional maps in both hemispheres. Typically, each callosal axon displayed 2 or 3 clusters of synaptic boutons in layer II/III and the upper part of layer IV. These clusters were preferentially distributed in regions representing the same orientation and the same visuotopic location as that at the corresponding injection sites in the opposite hemisphere. The spatial distribution of these clusters was elongated and its main axis correlated well with the preferred orientation at the injection site. These results demonstrate a specific organization of interhemispheric axons that link cortical regions representing the same orientation and the same location of visual stimuli. Visual callosal connections are thus likely involved in the processing of coherent information in terms of shape and position along the midline of the visual field, which may facilitate the fusion of both hemifields into the percept of a single visual scene.

Early antithrombotic treatment with warfarin oral suspension in severe neonatal protein C deficiency.

Neonatal severe protein C deficiency is a serious disease. There is no uniform approach for long-term preventive treatment of thrombotic events. We report the case of neonatal severe protein C deficiency treated with warfarin oral suspension. An international normalized ratio (INR) from 2.5 to 3.5 was expected. The INR was measured by home monitoring using the Coaguchek XS® (Roche Diagnostics, Mannheim, Germany) monitor. During 2years of warfarin treatment, there were only two minor episodes of purpuric access and no bleeding was reported. This case suggests that the early introduction of warfarin oral suspension, home-care monitoring, and parental education programs may be a beneficial treatment option for children with protein C deficiency.

Postnatal development of GFAP, connexin43 and connexin30 in cat visual cortex.

In cat visual cortex, neurons acquire progressively mature functional properties during the first postnatal months. The aim of this study was to analyze the development of astrocytes during this period. The patterns of expression of the glial fibrillary acidic protein (GFAP) as well as of two gap junction proteins expressed in astrocytes, connexin43 (Cx43) and connexin30 (Cx30), were investigated by immunohistochemistry and optical density measurements, in visual cortical areas 17 and 18 at four different ages: 2 weeks (postnatal days 12 to 15, P12-15), 1 month (P27-31), 2 months (P60-62) and beyond 1 year. Since visual experience is a key factor for neural development, the patterns of expression of these three proteins were studied both in normally-reared and monocularly deprived animals. Interestingly, the distribution of GFAP, Cx43 and Cx30 was found to change dramatically but independently of visual experience, during postnatal development, even beyond P60. During the first postnatal month, GFAP and Cx43 were mainly localized in the white matter underlying the visual cortical areas 17 and 18. Then, their distributions evolved similarly with a progressive decrease of their density in the white matter associated with an increase in the cortex. Connexin30 expression appeared only from the second postnatal month, strictly in the cortex and with a laminar distribution which was similar to that of Cx43 at the same age. In adults, a specific laminar distribution was observed, that was identical for GFAP, Cx43 and Cx30: their density was higher in layers II/III and V than in the other cortical layers.

Effects of afferent signals from the extraocular muscles upon units in the cerebellum, vestibular nuclear complex and oculomotor nucleus of the trout.

The responses of single units in the cerebellum, the vestibular nuclear complex and adjacent regions of the brainstem and in the oculomotor nucleus were studied in decerebrate, paralysed rainbow trout (Salmo gairdneri). Natural vestibular stimulation was provided by horizontal, sinusoidal oscillation of the fish and extraocular muscle afferents of the eye ipsilateral to the recording were activated either by passive eye-movement or by electrical stimulation of the trochlear (IV) nerve in the orbit. Unit responses to vestibular and/or orbital stimuli were examined in peristimulus-time histograms interleaved in time. In the cerebellum and brainstem, of 124 units exposed to both types of stimulus, 26 (21%) responded only to vestibular input, 26 (21%) were affected only by the orbital signal and 23 (18%) received both signals. The remaining 49 units (39%) responded to mechanical stimulation of the head or body or to vibration; they were labelled "polymodal" and discarded. The recording sites of 56 units were verified by histology; 30 were in the cerebellum and 26 in the brainstem. Input from the eye muscles had excitatory or inhibitory effects upon the vestibular responses. The effects of the orbital signal were usually phasic but rare tonic responses also occurred. About half (15 of 34) of the units which responded to passive eye-movement showed statistically significant differences in the magnitude of their responses to horizontal and to vertical eye-movement. More units preferred horizontal movement (11) than preferred vertical passive eye-movement (four). Note that the plane of vestibular stimulation was always horizontal. In the region of the oculomotor nucleus, of 19 units, five (26%) gave vestibular responses only and three (16%) were affected only by the orbital signal; three units (16%) with polymodal responses were discarded. Of the eight units carrying both signals, histological confirmation that the recording site lay in the column of cells forming the oculomotor/trochlear nuclei was obtained in four. The responses and interactions were similar to those found in the brainstem. The results present two principal points of interest. 1. They reinforce the accumulating body of evidence that, in species with widely different oculomotor and visual behaviour, signals from extraocular muscle proprioceptors reach the vestibulo-ocular system; this, in turn, suggests that these signals may play some rather fundamental role in the oculomotor system.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent signals from cat extraocular muscles in the medial vestibular nucleus, the nucleus praepositus hypoglossi and adjacent brainstem structures.

The responses of single units in the vestibular nuclei, nucleus praepositus hypoglossi and in the brainstem, deep and posterior to the abducens nucleus, were studied in anaesthetized, paralysed cats. Natural vestibular stimulation was provided by horizontal, sinusoidal oscillation of the animal and extraocular muscle afferents of the ipsilateral eye were activated either by passive eye-movement or by electrical stimulation of the inferior oblique branch of the oculomotor nerve in the orbit. Unit responses to vestibular and/or orbital stimuli were examined in sets of peristimulus time histograms interleaved in time. Of 127 units exposed to both types of stimulus, 40 (32%) responded only to vestibular input; 46 (32%) were affected only by the orbital afferent signal and 19 (15%) received both signals; the remaining 22 units (17%) were discarded because they had polymodal (usually somaesthetic) input. Of the 93 units whose recording sites were determined histologically, 24 were in the medial vestibular nucleus, 16 in the n. praepositus hypoglossi and 45 in the magnocellular nucleus of the reticular formation posterior and deep to the abducens nucleus. In these three nuclei 19 units in total were found which carried the orbital proprioceptive afferent signal and also responded to horizontal vestibular stimulation. The input from the eye muscles proved able to modify the vestibular response by adding excitation or inhibition or both. Effects of the orbital signal were generally phasic. About half of the units which responded to passive eye-movement showed statistically significant differences between their responses to horizontal and to vertical eye-movement. We have shown previously that signals from extraocular muscle proprioceptors reach the vestibulo-oculomotor system in an amphibian and a bony fish; the present experiments show that this is the case in a mammal also. The fact that the visual and visuomotor behaviour of these three species is very different suggests that the proprioceptive signal may play some rather fundamental role in the vestibulo-ocular system. The principal interest of the present results is that they demonstrate that units in the central vestibular system of the cat, in structures which are known to be concerned in oculomotor control, and particularly in the organization of horizontal eye-movement, receive an afferent signal from the eye muscles during passive eye-movement. These brainstem nuclei are known to receive various combinations of input from the vestibular and visual systems and of signals which represent neck movement and eye position and velocity.(ABSTRACT TRUNCATED AT 400 WORDS)

Area centralis position relative to the optic disc projection in kittens as a function of age.

Changes during development of the optic disc projection relative to the area centralis position in the visual field were studied in the kitten. The determinations were based both on direct measurement and physiological location of these retinal landmark projections. The results showed that the relative distance between these retinal landmarks in visual space became shorter with age, indicating that the visual field coordinates change extensively with age. Since the area centralis cannot be seen in most young kittens, the mean distances we have determined for the landmarks in the visual field may provide a useful means of estimating the position of area centralis from the projected position of the optic disc. Our results also confirm the nonuniform growth of the retina suggested already from anatomical observations. Taking into account both eye growth and the changes in the visual field coordinates allowed some reinterpretation of changes in physiological properties of the visual cells known to occur during development, such as decreased size of the visual receptive fields, increased spatial resolution and increased responsiveness to high velocity visual stimuli.

Visual inter-hemispheric processing: constraints and potentialities set by axonal morphology.

The largest bundle of axonal fibers in the entire mammalian brain, namely the corpus callosum, is the pathway through which almost half a billion neurons scattered over all neocortical areas can exert an influence on their contralateral targets. These fibers are thus crucial participants in the numerous cortical functions requiring collaborative processing of information across the hemispheres. One of such operations is to combine the two partial cortical maps of the visual field into a single, coherent representation. This paper reviews recent anatomical, computational and electrophysiological studies on callosal connectivity in the cat visual system. We analyzed the morphology of individual callosal axons linking primary visual cortices using three-dimensional light-microscopic techniques. While only a minority of callosal axons seem to perform a strict 'point-to-point' mapping between retinotopically corresponding sites in both hemispheres, many others have widespread arbors and terminate into a handful of distant, radially oriented tufts. Therefore, the firing of a single callosal neuron might influence several cortical columns within the opposite hemisphere. Computer simulation was then applied to investigate how the intricate geometry of these axons might shape the spatio-temporal distribution of trans-callosal inputs. Based on the linear relation between diameter and conduction velocity of myelinated fibers, the theoretical delays required for a single action potential to reach all presynaptic boutons of a given arbor were derived from the caliber, g-ratio and length of successive axonal segments. This analysis suggests that the architecture of callosal axons is, in principle, suitable to promote the synchronous activation of multiple targets located across distant columns in the opposite hemisphere. Finally, electrophysiological recordings performed in several laboratories have shown the existence of stimulus-dependent synchronization of visual responses across the two hemispheres. Possible implications of these findings are discussed in the context of temporal tagging of neuronal assemblies.

Microglia and astrocytes may participate in the shaping of visual callosal projections during postnatal development.

In the adult cat, axons running through the corpus callosum interconnect the border between the visual cortical areas 17 and 18 (A17 and A18) of both hemispheres. This specific pattern emerges during postnatal development, under normal viewing conditions (NR), from the elimination of initially exuberant callosal projections. In contrast, if the postnatal visual experience is monocular from birth (MD), juvenile callosal projections are stabilised throughout A17 and A18. The present study aimed at using such a model in vivo to find indications of a contribution of glial cells in the shaping of projections in the developing CNS through interactions with neurones, both in normal and pathological conditions. As a first stage, the distribution and the morphology of microglial cells and astrocytes were investigated from 2 weeks to adulthood. Microglial cells, stained with isolectin-B4, were clustered in the white matter below A17 and A18. Until one month, these clustered cells displayed an ameboid morphology in NR group, while they were more ramified in MD animals. Their phenotype thus depends on the postnatal visual experience, which indicates that microglial cells may interact with axons of visual neurones. It also suggests that they may differentially contribute to the elimination and the stabilisation of juvenile exuberant callosal fibres in NR and MD animals respectively. Beyond one month, microglial cells were very ramified in both experimental groups. Astrocytes were labelled with a GFAP-antibody. The distributions of connexins 43 (Cx43) and 30 (Cx30), the main proteic components of gap junction channels in astrocytes, were also investigated using specific antibodies. Both in NR and MD groups, until 1 month, GFAP-positive astrocytes and Cx43 were mainly localised within the subcortical white matter. Then GFAP, Cx43 and Cx30 stainings progressively appeared within the cortex, throughout A17 and A18 but with a differential laminar expression according to the age. Thus, the distributions of both astrocytes and connexins changed with age; however, the monocular occlusion had no visible effect. This suggests that astrocytes may contribute to the postnatal development of neuronal projections to the primary visual cortex, including visual callosal projections.

Visual callosal connections and strabismus.

Strabismus is a condition that exists when the visual axes of the two eyes fail to intersect at the fixation point under binocular viewing conditions. When it occurs in mammals during the critical period which corresponds to the period of maximal plasticity early in life, strabismus is known to induce both morphological anomalies and abnormal connections from the retina to the cortex; it further leads to binocular neural changes and to spatial vision deficits, especially at the cortical level. After a brief review of the already known data about the consequences of early strabismus in cats, monkeys and humans on the development of the visual system and of visual perception, new data are presented here concerning interhemispheric connections in the cat. In normally-reared kittens, visual callosal transfer is shown to be almost adult-like as soon as 12 days after birth: it is almost limited to the 17/18 border of the visual cortex when using visual stimulations in spite of the presence of still numerous juvenile exuberant callosal projections. In contrast, callosal transfer of visual information is extended to both areas 17 and 18 after strabismus, leading to the conclusion that at least some juvenile exuberant callosal projections are not only anatomically but also functionally stabilized after such an oculomotor disease. The possibility that similar abnormalities might be present in monkeys and humans is discussed.

Role of eye movements in developmental processes of orientation selectivity in the kitten visual cortex.

Six-week-old dark-reared kittens were exposed for 6 hr to a normal lighted environment in which they were free to move, but with eyes immobilized. Receptive field properties (orientation selectivity and ocular dominance) of the visual cortical cells (Area 17) were studied. No restoration of orientation selectivity could be observed when only the eyes were immobilized either by oculomotor nerve sections or by extraocular muscles removal, in contrast to what had been observed in intact free moving animals. From these results one can conclude that eye movements must be associated with vision to allow developmental processes of orientation selectivity in the primary cortex. These new results are compared to those obtained previously in paralyzed or restrained animals and those obtained in animals with interruption of orbital afferents.

Layout of transcallosal activity in cat visual cortex revealed by optical imaging.

The contribution of interhemispheric connections to functional maps in cat visual cortex was investigated by using optical imaging of intrinsic signals. In order to isolate the functional inputs arriving via the corpus callosum (CC) from other inputs, we used the split-chiasm preparation. The regions activated through the CC in visual areas 17 (A17) and 18 (A18) were localized and characterized by stimulating monocularly split-chiasm cats with moving, high contrast oriented gratings. We found that the CC mediates the activation of orientation selective domains in the transition zone (TZ) between A17 and A18 and occasionally within portions of both of these areas. We observed transcallosally activated orientation domains all along the TZ without any obvious interruption, and these domains were arranged around "pinwheel" centers. Interestingly, the TZ was divided in two parallel regions, which resemble A17 and A18 in their preferred temporal and spatial frequencies. Finally, we demonstrated that orientation maps evoked through the transcallosal and geniculo-cortical pathways were similar within the TZ, indicating a convergence of inputs of matching orientations in this region. These results contribute to a better understanding of the role of the CC in visual perception of orientations and shapes, at the level of the visual cortex.

Unilateral paralytic strabismus in the adult cat induces plastic changes in interocular disparity along the visual midline: contribution of the corpus callosum.

Neurones activated through the corpus callosum (CC) in the cat visual cortex are known to be almost entirely located at the 17/18 border. They are orientation selective and display receptive fields (RFs) distributed along the central vertical meridian of the visual field ("visual midline"). Most of these cells are binocular, and many of them are activated both from the contralateral eye through the CC, and from the ipsilateral eye via the direct retino-geniculo-cortical (GC) pathway. These two pathways do not carry exactly the same information, leading to interocular disparity between pairs of RFs along the visual midline. Recently, we have demonstrated that a few weeks of unilateral paralytic strabismus surgically induced at adulthood does not alter the cortical distribution of these units but leads to a loss of their orientation selectivity and an increase of their RF size, mainly toward the ipsilateral hemifield when transcallosally activated (Watroba et al., 2001). To investigate interocular disparity, here we compared these RF changes to those occurring in the same neurones when activated through the ipsilateral direct GC route. The 17/18 transition zone and the bordering medial region within A17 were distinguished, as they display different interhemispheric connectivity. In these strabismics, some changes were noticed, but were basically identical in both recording zones. Ocular dominance was not altered, nor was the spatial distribution of the RFs with respect to the visual midline, nor the amplitude of position disparity between pairs of RFs. On the other hand, strabismus induced a loss of orientation selectivity regardless of whether neurones were activated directly or through the CC. Both types of RFs also widened, but in opposite directions with respect to the visual midline. This led to changes in incidences of the different types of position disparity. The overlap between pairs of RFs also increased. Based on these differences, we suggest that the contribution of the CC to binocular vision along the midline in the adult might be modulated through several intrinsic cortical mechanisms.

[Dissymmetry in the interhemispheric transfer between visual areas in the adult cat, induced by chiasmotomy and monocular occlusion]. / Dissymétrie du transfert interhémisphérique entre aires visuelles chez le chat adulte, induite par chiasmotomie et occlusion monoculaire.

In the adult cat, a midsagittal section of the optic chiasm, which deprives each hemisphere of about half of its visual afferences, is followed after 6 weeks by a change in the callosal interhemispheric transfer, if one eye was occluded during the whole postoperative recovery period, thus depriving one hemisphere of direct visual messages related to patterned vision: with respect to controls only chiasmotomized, with binocular visual experience during an identical postoperative period, the deprived hemisphere displayed a significantly larger callosal afferent traffic while its own callosal efferences were on the contrary significantly reduced. This dissymmetry reveals that important postoperative changes can thus take place even in the adult animal.

[Reorganization of callosal interhemispheric connections in the adult cat: effects of monocular occlusion after chiasmotomy]. / Réorganisation des connexions interhémisphériques calleuses chez le chat adulte: effets de l'occlusion monoculaire après chiasmotomie.

Two groups of adult cats were chiasmotomized and their cortical receptive fields (17-18 boundary) were compared after a postoperative period of ca 6 weeks. In one group, binocular vision was maintained during that period, in the other one, one eye was sutured at the time of the chiasmotomy, depriving one hemisphere from patterned vision through the direct pathway. In monocular chiasmotomized animals, the receptive fields to stimulation of the contralateral eye were significantly larger than in the binocular ones.

[Responsive characteristics of the cells of cortex area 18 of the adult cat after chiastomy. Postoperative development; importance of the visual experience]. / Caractéristiques réactionnelles des cellules de l'aire corticale 18 du chat adulte après chiasmotomie. Evolution postopératoire; importance de l'expérience visuelle.

Patterns of unit responses in area 18 to stimulation of the ipsilateral temporal retina were studied in adult Cats at different times after complete midsagittal section of the optic chiasm. A specific postoperative evolution could thus be followed. Immediately after section, the percentage of cells reacting to light was reduced, as compared to control preparations, as well as the average areas of the receptive fields, owing to the disappearance of large eccentric fields. A progressive recovery of the normal reactive patterns occurred between 8 and 45 days after chiasmotomy, provided that the animal was placed in normal visual conditions during its postoperative period. No such recovery could be observed after 55 days, in animals kept in complete darkness after operation. Visual experience thus seems essential to this kind of functional recovery in the adult. Another characteristic of chiasmotomised Cats also appeared to depend upon postoperative visual experience, namely the development of cells with "diffuse" responses to light.

Receptive field sizes and responsiveness to light in area 18 of the adult cat after chiasmotomy. Postoperative evolution; role of visual experience.

Receptive field sizes to stimulation of the ipsilateral temporal retina were studied in area 18 of adult cats at different times after complete midsagittal section of the optic chiasm. A specific postoperative evolution could thus be noticed: immediately after section, the average area of the receptive fields was reduced, as compared to control preparations, owing to the disappearance of large fields located at more than 20 degrees of eccentricity. A progressive reappearance of these large fields occurred between 8 and 45 days after chiasmotomy, provided that the animal was placed in normal visual conditions during its postoperative period. No such recovery could be assessed after as long as 55 days, in animals kept in complete darkness after operation. Chiasmotomized cats also displayed a reduction of their percentage of light reactive cells with respect to controls, as expected from the suppression of the contralateral input. This percentage was at first very low and progressively increased, during postoperative recovery but again not when the animal had been kept in the dark. Finally, an increase of cells with "diffuse responses" was observed in the late postoperative recovery stage. This latter evolution also appeared to depend upon postoperative visual experience. On the other hand, no clear indication of an interhemispheric transfer could be obtained in these experiments, even at the 17-18 boundary.

Visual interhemispheric transfer to areas 17 and 18 in cats with convergent strabismus.

Commissural connections between primary visual cortical maps of the two hemispheres are essential to unify the split representation of the visual field. In normal adult cats, callosal connections are essentially restricted to the border between areas A17 and A18, where the central vertical meridian is projected. In contrast, early convergent strabismus leads to an expanded callosal-receiving zone, as repeatedly indicated by anatomical experiments. We investigated here the functional correlates of this widespread distribution of callosal terminals by analysing transcallosal visual responses in five anaesthetized and paralysed 4-10-month-old cats whose right eye had been surgically deviated on postnatal day 6. After acute section of the optic chiasm, single-unit activity was recorded from A17 and A18 of the right hemisphere while the left eye was visually stimulated. A total of 108/406 units were transcallosally activated. While they were more frequent at the 17/18 border (46% of the units recorded within this region), numerous transcallosally activated units were located throughout A17 (16%), A18 (27%) or within the white matter (17%). In all regions, transcallosally driven units displayed functional deficits usually associated with strabismus, such as decreased binocularity and ability to respond to fast-moving stimuli, and increased receptive field size. Many units also displayed reduced orientation selectivity and increased position disparity. In addition, transcallosal receptive fields were manifestly located within the hemifield ipsilateral to the explored cortex, with almost no contact with the central vertical meridian. Comparison with data from normal adults revealed that strabismus induced a considerable expansion of the callosal receiving zone, both in terms of the cortical region and of the extent of the visual field involved in interhemispheric transfer, with implications in the integration of visual information across the hemispheres.

Impairment of binocular vision in the adult cat induces plastic changes in the callosal cortical map.

In the primary visual cortex of normally reared adult cat, neurons activated through the corpus callosum are almost entirely located at the 17/18 border. They display small receptive fields distributed along the central vertical meridian of the visual field and are orientation selective. Here we demonstrate that a few weeks of monocular deprivation or unilateral convergent strabismus produced in adulthood does not modify the cortical distribution of these neurons, but leads to an increase of their receptive field size mainly toward the ipsilateral hemifield and to a loss of their orientation selectivity. We conclude that manipulation of binocular vision in the adult modifies neither the location of the primary callosal cortical map nor its retinotopy. In contrast, it induces functional plastic changes in this map which lead to a significant widening of the area of visual space signalled through the corpus callosum. These plastic changes are interpreted as the result of the strengthening of normally hidden subthreshold synaptic inputs.