Distant heterotopic callosal connections to premotor cortex in non-human primates.

Cortico-cortical connectivity has become a major focus of neuroscience in the last decade but most of the connectivity studies focused on intrahemispheric circuits. Little has been reported about information acquired and processed in the premotor cortex and its functional connection with its homotopic counterpart in the opposite hemisphere via the corpus callosum. In non-human primates (macaques) lateralization is not well documented and its exact role is still unknown. The present study confirms in two macaques the existence of homotopic contralateral projections and completes the picture by further exploring heterotopic (non-motor) callosal projections. This was tested by injecting retrograde tracers in the premotor cortical areas PMv and PMd (targets). Our method consisted of identifying the connections with all the homo- and heterotopic cortical areas located in the contralateral hemisphere. The results showed that PMd and PMv receive multiple low-density labeled inputs from the opposite heterotopic prefrontal, parietal, motor, insular and temporal regions. Such unexpected collection of transcallosal inputs from heterotopic areas suggests that the premotor areas communicate with other modalities through long distance low-density networks which could have important implications in the understanding of sensorimotor and multimodal integration.

Effects of dorsolateral prefrontal cortex lesion on motor habit and performance assessed with manual grasping and control of force in macaque monkeys.

In the context of an autologous adult neural cell ecosystem (ANCE) transplantation study, four intact adult female macaque monkeys underwent a unilateral biopsy of the dorsolateral prefrontal cortex (dlPFC) to provide the cellular material needed to obtain the ANCE. Monkeys were previously trained to perform quantitative motor (manual dexterity) tasks, namely, the "modified-Brinkman board" task and the "reach and grasp drawer" task. The aim of the present study was to extend preliminary data on the role of the prefrontal cortex in motor habit and test the hypothesis that dlPFC contributes to predict the grip force required when a precise level of force to be generated is known beforehand. As expected for a small dlPFC biopsy, neither the motor performance (score) nor the spatiotemporal motor sequences were affected in the "modified-Brinkman board" task, whereas significant changes (mainly decreases) in the maximal grip force (force applied on the drawer knob) were observed in the "reach and grasp drawer" task. The present data in the macaque monkey related to the prediction of grip force are well in line with the previous fMRI data reported for human subjects. Moreover, the ANCE transplantation strategy (in the case of stroke or Parkinson's disease) based on biopsy in dlPFC does not generate unwanted motor consequences, at least as far as motor habit and motor performance are concerned in the context of a sequential grasping a small objects, which does not require the development of significant force levels.

Evolution of anaesthesia care and related events between 1996 and 2010 in Switzerland.

BACKGROUND: Anaesthesia Databank Switzerland (ADS) is a voluntary data registry introduced in 1996. Its ultimate goal is to promote quality in anaesthesiology. METHODS: The ADS registry analyses routinely recorded adverse events and provides benchmark comparisons between anaesthesia departments. Data collection comprises a set of 31 variables organised into three modules, one mandatory and two optional. RESULTS: In 2010, the database included 2,158,735 anaesthetic procedures. Over time, the proportions of older patients have increased, the largest group being aged 50-64 years. The percentage of patients with American Society of Anesthesiologists (ASA) status 1 has decreased while the percentage of ASA status 2 or 3 patients has increased. The most frequent comorbidities recorded were hypertension (21%), smoking (16%), allergy (15%) and obesity (12%). Between 1996 and 2010, 125,579 adverse events were recorded, of which 34% were cardiovascular, 7% respiratory, 39% technical and 20% non-specific. The most severe events were resuscitation (50%), oliguria (22%), myocardial ischaemia (17%) and haemorrhage (10%). CONCLUSION: Routine ADS data collection contributes to the monitoring of trends in anaesthesia care in Switzerland. The ADS system has proved to be usable in daily practice, although this remains a constant challenge that is highly dependent on local quality management and quality culture. Nevertheless, success in developing routine regular feedback to users to initiate discussions about anaesthetic events would most likely help strengthen departmental culture regarding safety and quality of care.

Refined methodology for implantation of a head fixation device and chronic recording chambers in non-human primates.

The present study was aimed at developing a new strategy to design and anchor custom-fitted implants, consisting of a head fixation device and a chronic recording chamber, on the skull of adult macaque monkeys. This was done without the use of dental resin or orthopedic cement, as these modes of fixation exert a detrimental effect on the bone. The implants were made of titanium or tekapeek and anchored to the skull with titanium screws. Two adult macaque monkeys were initially implanted with the head fixation device several months previous to electrophysiological investigation, to allow optimal osseous-integration, including growth of the bone above the implant's footplate. In a second step, the chronic recording chamber was implanted above the brain region of interest. The present study proposes two original approaches for both implants. First, based on a CT scan of the monkey, a plastic replicate of the skull was obtained in the form of a 3D print, used to accurately shape and position the two implants. This would ensure a perfect match with the skull surface. Second, the part of the implants in contact with the bone was coated with hydroxyapatite, presenting chemical similarity to natural bone, thus promoting excellent osseous-integration. The longevity of the implants used here was 4 years for the head fixation device and 1.5 years for the chronic chamber. There were no adverse events and daily care was easy. This is clear evidence that the present implanting strategy was successful and provokes less discomfort to the animals.

New N(4)-substituted piperazine naphthamide derivatives as BACE-1 inhibitors.

Synthesis and enzymatic evaluation of new series of N(4)-substituted piperazine naphthamide derivatives as BACE-1 inhibitors for the treatment of Alzheimer's disease are reported.

Origin of thalamic inputs to the primary, premotor, and supplementary motor cortical areas and to area 46 in macaque monkeys: a multiple retrograde tracing study.

The origin of thalamic inputs to distinct motor cortical areas was established in five monkeys to determine whether the motor areas receive inputs from a common thalamic nucleus and the extent to which the territories of origin overlap. To not rely on the rough definition of cytoarchitectonic boundaries in the thalamus, monkeys were subjected to multiple injections of tracers (four to seven) in the primary (M1), premotor (PM), and supplementary (SMA) motor cortical areas and in area 46. The cortical areas were distributed into five groups, each receiving inputs from a specific set of thalamic nuclei: 1) M1; 2) SMA-proper and the caudal part of the dorsal PM (PMdc); 3) the rostral and caudal parts of the ventral PM (PMvr and PMvc); 4) the rostral part of the dorsal PM (PMdr); and 5) the superior and inferior parts of area 46 (area 46sup and area 46inf). A major degree of overlap was obtained for the origins of the thalamocortical projections directed to areas 46inf and 46sup and for those terminating in SMA-proper and PMdc. PMvc and PMvr received inputs from adjacent and/or common thalamic regions. In contrast, the degree of overlap between M1 and SMA was smaller. The projection to M1 shared relatively limited zones of origin with the projections directed to PM. Thalamic inputs to the motor cortical areas (M1, SMA, PMd, and PMv), in general, were segregated from those directed to area 46, except in the mediodorsal nucleus, in which there was clear overlap of the territories sending projections to area 46, SMA-proper, and PMdc.

Dexterity in adult monkeys following early lesion of the motor cortical hand area: the role of cortex adjacent to the lesion.

Infant monkeys were subjected to unilateral lesions of the motor cortex (mainly its hand representation). After maturation, they showed normal use of the contralateral hand for global grip movements. However, as compared with the ipsilateral hand, precision grip tasks requiring relatively independent finger movements were performed with less dexterity, particularly if adjustments of the wrist position were necessary. The purpose of this study was to investigate mechanisms which may be responsible for the rather well, although not complete, preservation of manipulative behaviour of these adult monkeys. To this end, the hand representations were mapped bilaterally with intracortical microstimulation in the mature monkeys, and the dexterity of both hands assessed quantitatively in a precision grip task. The behavioural effects of reversible inactivations of the primary (M1) and supplementary (SMA) motor cortical areas were then tested. The following were found. (i) The hand contralateral to the lesion exhibited subtle but significant dexterity deficits, as compared with the ipsilateral hand; the deficit was essentially for complex movements requiring dissociation of the thumb-index finger pinch from the other digits, involving also an arm rotation. (ii) Reversible inactivation of the M1 hand representation in the intact hemisphere dramatically impaired dexterity of the opposite hand without affecting the ipsilateral hand (contralateral to the early lesion). (iii) A relatively complete hand representation was found to occupy a new territory, medial to the old lesion. (iv) The role of this new displaced representation was crucial for the preserved dexterity of the opposite hand, as evidenced by its functional inactivation. In contrast, inactivation of both SMA cortices did not interfere with the manipulative behaviour. It is thus concluded that the preserved functional capacity of manipulations with the hand opposite the early lesion can be essentially attributed to a cortical reorganization around the old lesion. Under the present experimental conditions, contributions from either the SMA or the intact M1 appear not to be crucial.

Antibody-dependent Fos-like immunoreactivity (FLI) in the auditory pathway of the rat in response to electric stimulation of the cochlea.

The distribution of Fos-like immunoreactivity (FLI) in the auditory pathway was analyzed and quantified in two groups of rats subjected to a similar electric stimulation of the cochlea, but the brains of the two groups were treated with a different antibody against Fos. The brains of control (unstimulated) rats were processed and analyzed in the same way. In a given auditory nucleus, for the same paradigm of stimulation, a very different density of FLI can be found depending on the antibody used to process the tissue. There are auditory nuclei which exhibited a significant activation of FLI as compared to control animals when one antibody was used whereas there was no difference when using the other antibody. In other cases, for both antibodies, a significant FLI increase was observed in relation to the stimulus. However, the increase of FLI density was significantly greater in some auditory nuclei with one antibody, while it was the reverse in other auditory nuclei. The present data demonstrate quantitatively that the use of two different Fos antibodies can strongly affect the FLI produced in the auditory pathway by electric stimulation of the cochlea, confirming previous qualitative observations based on acoustic stimulation of the cochlea.

Dual morphology and topography of the corticothalamic terminals originating from the primary, supplementary motor, and dorsal premotor cortical areas in macaque monkeys.

In the motor, somatosensory, and auditory systems of rodents and cats, the corticothalamic connection is composed of a main projection formed by small endings and a minor projection terminating with giant endings. To establish whether the corticothalamic projection originating from motor cortical areas in primates exhibits the same duality, the anterograde tracer biotinylated dextran amine was injected in eight macaque monkeys in the primary motor (M1; n = 3), the supplementary motor (SMA; n = 3) and the dorsal premotor (PMd; n = 2) cortical areas to label corticothalamic axons. The corticothalamic projection originating from these three motor cortical areas was characterized by the presence of axon terminals constituting the same two types of endings, observed both as boutons en passant and terminaux. The population of small endings exhibited a mean cross-sectional maximum diameter of 0.95 microm (S.D. = 0.23), a range of diameters not overlapping that of giant endings (mean diameter = 3.46 microm, S.D. = 0.74 microm). Topographically, the giant endings originating from M1 were located in the same thalamic nucleus (ventroposterolateral nucleus, oral part) in which the small endings were found. In contrast, the giant endings originating from SMA and PMd were located in a thalamic nucleus (mediodorsal nucleus) distinct from the main termination zone formed by small endings. Along the rostrocaudal axis, the giant endings were distributed in a restricted zone, irrespective of the origin of the projection (M1, SMA, PMd). The dual morphology of corticothalamic endings, previously found in rodents and cats, is present in the motor system of subhuman primates for both primary and nonprimary motor cortical areas.

The Lyn-catalyzed Tyr phosphorylation of the transmembrane band-3 protein of human erythrocytes.

Band-3 protein (approximately 95 kDa), the major and multifunctional transmembrane protein of human erythrocytes, has been shown to be phosphorylated by endogenous Tyr-protein kinases on different Tyr residues at its N and C cytoplasmic domains. Both the added p36syk (catalytic domain of p72syk) and Lyn kinases are able to phosphorylate the isolated cytoplasmic domain of band 3 (cdb3), yielded by chymotryptic digestion of band 3 in the isolated membranes (ghosts). However, the two Tyr-protein kinases exhibited different phosphorylation behaviours when added to the isolated erythrocyte membranes. More precisely, the added p36syk markedly Tyr phosphorylates the band-3 protein, whereas the added Lyn phosphorylates it very poorly. It is of interest that Lyn can associate with membranes and markedly phosphorylate band 3 when this latter protein has been previously phosphorylated by p36syk, i.e. the p36(syk)-catalyzed phosphorylation is proposed to be a prerequisite for the association of Lyn with the membrane (likely to band 3) and for the Lyn-catalyzed phosphorylation of different band-3 Tyr sites.

Evidence for direct connections between the hand region of the supplementary motor area and cervical motoneurons in the macaque monkey.

In primates the corticospinal neurons of the hand representation of the primary motor cortex (M1) give rise to direct contacts with the cervical motoneurons that control distal forelimb muscles. We investigated, at the light-microscopy level, whether corticospinal cells present in the hand area of the supplementary motor area (SMA) also establish direct connections with cervical motoneurons, particularly those innervating hand and finger muscles. The hand representation of the M1 (two monkeys) or SMA (two monkeys) was located using intracortical microstimulation and injected with the anterograde tracer biotinylated dextran amine to label corticospinal terminals. Forearm muscles acting on the wrist and hand as well as hand muscles acting on the thumb and index finger, thus including those activated by intracortical stimulation, were injected with the retrograde tracer cholera-toxin B subunit, in order to label the motoneurons. A consistent zone of overlap between the two markers was found in the cervical cord. Close appositions between corticospinal axonal terminals and the somata or dendrites of motoneurons were found after injection in the M1, confirming previous observations. The new finding is the observation of similar close appositions after injection in the SMA, suggesting its control of hand movements in parallel with the M1.

Electrically induced fos-like immunoreactivity in the auditory pathway of the rat: effects of survival time, duration, and intensity of stimulation.

The goal of the present study was to establish how Fos-like immunoreactivity (FLI) elicited in the rat auditory pathway by unilateral electric stimulation of the cochlea is affected by the following experimental parameters: duration and intensity of stimulation, duration of survival time after offset of stimulation. The dense FLI found in the ipsilateral dorsal cochlear nucleus, as well as the moderate FLI found in the contralateral dorsal cochlear nucleus and in the posteroventral cochlear nucleus on both sides, were consistent after survival times ranging from 0 to 2-3 h, but they significantly decreased after longer survival times (5 and 6 h). In the same nuclei, FLI was increased even by short durations of stimulation (5 and 10 min) as compared to control rats, although FLI progressively increased for longer stimulation (20 and 45 min). In the auditory thalamus, FLI was found mainly in the peripeduncular nucleus, the dorsal and medial divisions of the medial geniculate body, whereas its ventral division was virtually devoid of immunoreactive neurons. This pattern of FLI distribution in the auditory thalamus persisted even after relatively long survival times (5 and 6 h). In both the cochlear nucleus and auditory thalamus, the density of FLI slightly increased in parallel with the intensity of stimulation. In other auditory nuclei, such as the inferior colliculus and the nucleus of the lateral lemniscus, there was no simple relation between the density of FLI and the three tested experimental parameters. Thus, the distribution and density of FLI did not vary in parallel in the various nuclei of the auditory pathway as a function of the tested experimental parameters; different patterns of FLI changes were instead observed in different auditory nuclei.

Functional link between phosphorylation state of membrane proteins and morphological changes of human erythrocytes.

The Tyr-phosphorylation of the cytoplasmic domain of the major membrane-spanning band 3, rather than the Ser/Thr-phosphorylation of the membrane proteins (spectrin and band 3 itself), might be functionally related to certain morphological changes of human erythrocytes. This view is supported by the following lines of evidence: a) vanadate or its derivative pervanadate (vanadyl hydroperoxide), which markedly increase the Tyr-phosphorylation of band 3 (without practically affecting the Ser/Thr-phosphorylation of spectrin) promotes a crenation of human erythrocytes; b) okadaic acid, which selectively increases the Ser/Thr-phosphorylation of spectrin and other membrane proteins, does not promote any shape change, at least at a level detectable with scanning electron microscopy.

Re-examination of the plasticity of the corticothalamic projection after unilateral neonatal lesion of the sensorimotor cortex in the rat: a phaseolus vulgaris-leucoagglutinin tracing study.

After unilateral lesion of the sensorimotor cortex in neonatal rats (postnatal days 2-4), the reorganization of the corticothalamic projection originating from the opposite intact sensorimotor cortex was studied. Three months after the lesion, the anterograde tracer phaseolus vulgaris-leucoagglutinin (PHA-L) was injected in the caudal forelimb area (CFA) of the intact opposite sensorimotor cortex to label the corticothalamic axons, in order to establish their trajectory as well as the distribution of their terminal fields in the thalamus. As previously reported, in addition to the standard ipsilateral corticothalamic projection, lesioned rats were characterized also by the presence of a contralateral corticothalamic projection, reaching the decorticate hemithalamus. There is some controversy in literature whether such bilateral corticothalamic projection is present in normal intact rats. On the basis of counts of axons crossing the midline to innervate the hemithalamus contralateral to the cortical injection, the present study provides evidence that the corticothalamic projection is indeed bilateral in normal rats, although the contralateral projection is sparse. Lesioned rats subjected to a similar small size PHA-L injection in the CFA of the intact opposite cortex showed a significantly increased contralateral corticothalamic projection. The number of labeled axons crossing the midline in the thalamus was, on average, five times higher in lesioned rats than in normal rats. It can be concluded that the contralateral corticothalamic projection from the CFA, observed after neonatal lesion of the opposite sensorimotor cortex, is not a novel projection resulting from the lesion, but rather an enhancement and extension of an existing, minor, projection present in normal rats. The enhancement of the crossing corticothalamic projection in lesioned rats is due to an increase of the number of crossing axons in the thalamus rather than to an increase of the degree of branching of the few axons present in control animals.

Spermine effect on the binding of casein kinase I to the rat liver mitochondrial structures.

The results indicated here, together with those previously reported, show that spermine, ubiquitous polyamine, while promoting the transmembrane translocation of casein kinase II (CKII) across the outer membrane to more internal compartments of rat liver mitochondria, promotes the binding of casein kinase I (CKI) to the external surface of outer mitochondrial membrane but inhibits its spontaneously occurring binding to more internal structures.

Relationship between membrane protein phosphorylation and intracellular translocation of casein kinase in human erythrocytes.

The present paper shows that an increased phosphorylation of the membrane proteins, promoted by the okadaic acid (strong inhibitor of P-Ser/Thr-protein phosphatase(s)), is accompanied by a release of casein kinase from the membrane into cytosol. Such an intracellular translocation might provide a feedback mechanism for the regulation of the casein kinase catalyzed phosphorylation of membrane proteins in the human erythrocytes.

Cerebellothalamocortical and pallidothalamocortical projections to the primary and supplementary motor cortical areas: a multiple tracing study in macaque monkeys.

The goal of the present study was to clarify whether the primary motor cortex (M1) and the supplementary motor cortex (SMA) both receive, via the motor thalamus, input from cerebellar and basal ganglia output nuclei. This is the first investigation that explores the problem by direct comparison, in the same animal, of thalamic zones that 1) project to M1 and SMA and 2) receive cerebellar-nuclear (CN) and pallidal (GP) afferents. These four zones were mapped in two monkeys by means of two retrograde tracers for M1 and SMA injections and of two anterograde tracers for CN and GP injections. All injections were performed under electrophysiological control (microstimulation and multiunit recordings). Injections in cortical areas were restricted to the hand/arm representation; in the SMA, the tracer deposit was within the "SMA-proper" (or "area F3") and did not include its rostral extension ("pre-SMA" or "area F6"). It was found that zones of all four types formed a number of highly complex patches of labeling that were usually not confined to one cytoarchitectonically defined thalamic nucleus. The overlap of clusters of labeled terminals and perikarya was evaluated morphometrically (area measurements) on a number of coronal sections along the anteroposterior extent of the motor thalamus. In line with previous studies, the thalamic territories innervated by CN and GP afferents rarely overlapped. However, zones projecting to M1 and/or to SMA included thalamic regions receiving CN as well as GP projections, providing the first evidence of such overlap from individual animals. The present observations support the previous conclusion from this laboratory (based on transsynaptic labeling) that the SMA receives, apart from its strong pallidal transthalamic input, a CN transthalamic input. These present findings that both M1 and SMA are recipients of transthalamic inputs from GP and CN thus support the concept that a mixed subcortical input consisting of weighted contributions from cerebellum, basal ganglia, substantia nigra, and spinothalamic tract is directed to each functional component of the sensorimotor cortex.

Spermine-mediated casein kinase II-uptake by rat liver mitochondria.

Spermine, ubiquitous intracellular polyamine, is able to promote the transmembrane translocation of casein kinase CKII through the outer membrane of rat liver mitochondria and its binding to more internal mitochondrial structures. These findings suggest that spermine may play a critical role in regulating the subcellular distribution of casein kinase CKII.

Transcallosal connections of the distal forelimb representations of the primary and supplementary motor cortical areas in macaque monkeys.

The goal of the present neuroanatomical study in macaque monkeys was twofold: (1) to clarify whether the hand representation of the primary motor cortex (M1) has a transcallosal projection to M1 of the opposite hemisphere; (2) to compare the topography and density of transcallosal connections for the hand representations of M1 and the supplementary motor area (SMA). The hand areas of M1 and the SMA were identified by intracortical microstimulation and then injected either with retrograde tracer substances in order to label the neurons of origin in the contralateral motor cortical areas (four monkeys) or, with an anterograde tracer, to establish the regional distribution and density of terminal fields in the opposite motor cortical areas (two monkeys). The main results were: (1) The hand representation of M1 exhibited a modest homotopic callosal projection, as judged by the small number of labeled neurons within the region corresponding to the contralateral injection. A modest heterotopic callosal projection originated from the opposite supplementary, premotor, and cingulate motor areas. (2) In contrast, the SMA hand representation showed a dense callosal projection to the opposite SMA. The SMA was found to receive also dense heterotopic callosal projections from the contralateral rostral and caudal cingulate motor areas, moderate projections from the lateral premotor cortex, and sparse projections from M1. (3) After injection of an anterograde tracer (biotinylated dextran amine) in the hand representation of M1, only a few small patches of axonal label were found in the corresponding region of M1, as well as in the lateral premotor cortex; virtually no label was found in the SMA or in cingulate motor areas. Injections of the same anterograde tracer in the hand representation of the SMA, however, resulted in dense and widely distributed axonal terminal fields in the opposite SMA, premotor cortex, and cingulate motor areas, while labeled terminals were clearly less dense in M1. It is concluded that the hand representations of the SMA and M1 strongly differ with respect to the strength and distribution of callosal connectivity with the former having more powerful and widespread callosal connections with a number of motor fields of the opposite cortex than the latter. These anatomical results support the proposition of the SMA being a bilaterally organized system, possibly contributing to bimanual coordination.

Effect of okadaic acid on membrane protein phosphorylation in human erythrocytes.

Okadaic acid, penetrating the human erythrocytes, almost completely inhibits P-Ser-protein phosphatase activity, whereas it unaffects Ser/Thr-protein kinase activity (casein kinases CKI and CKII), thus promoting a marked increase of the endogenous Ser-phosphorylation level of membrane proteins, such as cytoskeletal spectrin beta-subunit (band 2) and transmembrane band 3 protein. By contrast, the Tyr-phosphorylation state of band 3 protein is practically unaffected by okadaic acid, being unaffected both Tyr-protein kinase and P-Tyr-protein phosphatase activities.