Dysregulation of the epigenome in human breast cancer: contributions of gene-specific DNA hypermethylation to breast cancer pathobiology and targeting the breast cancer methylome for improved therapy.

Triple-negative breast cancers (including basal-like and claudin-low molecular subtypes) represent 20% to 25% of all breast cancers, but disproportionately contribute to breast cancer-associated death. We have identified a novel fundamental biological property of triple-negative breast cancers: most triple-negative breast cancers express aberrant DNA hypermethylation due to overexpression of DNA methyltransferase 3b (and hyperactivity of the DNA methyltransferase enzymes). DNA methyltransferase 3b overexpression occurs secondary to loss of miRNA-mediated post-transcriptional regulation. The resulting hyperactivity of DNA methyltransferase 3b produces concurrent DNA methylation-dependent silencing of numerous critical gene targets (including tumor suppressors and pro-apoptotic genes) and resistance to cytotoxic chemotherapy. This observation presents new opportunities for development of innovative treatment strategies on the basis of the epigenome as a novel therapeutic target in triple-negative breast cancers. Epigenetic therapy represents a new principle in cancer treatment in which restoration of critical molecular pathways occurs secondary to reexpression of silenced genes that encode negative mediators of cancer cell growth.

Dysregulation of the epigenome in triple-negative breast cancers: basal-like and claudin-low breast cancers express aberrant DNA hypermethylation.

A subset of human breast cancer cell lines exhibits aberrant DNA hypermethylation that is characterized by hyperactivity of the DNA methyltransferase enzymes, overexpression of DNMT3b, and concurrent methylation-dependent silencing of numerous epigenetic biomarker genes. The objective of this study was to determine if this aberrant DNA hypermethylation (i) is found in primary breast cancers, (ii) is associated with specific breast cancer molecular subtypes, and (iii) influences patient outcomes. Analysis of epigenetic biomarker genes (CDH1, CEACAM6, CST6, ESR1, GNA11, MUC1, MYB, SCNN1A, and TFF3) identified a gene expression signature characterized by reduced expression levels or loss of expression among a cohort of primary breast cancers. The breast cancers that express this gene expression signature are enriched for triple-negative subtypes - basal-like and claudin-low breast cancers. Methylation analysis of primary breast cancers showed extensive promoter hypermethylation of epigenetic biomarker genes among triple-negative breast cancers, compared to other breast cancer subclasses where promoter hypermethylation events were less frequent. Furthermore, triple-negative breast cancers either did not express or expressed significantly reduced levels of protein corresponding to methylation-sensitive biomarker gene products. Together, these findings suggest strongly that loss of epigenetic biomarker gene expression is frequently associated with gene promoter hypermethylation events. We propose that aberrant DNA hypermethylation is a common characteristic of triple-negative breast cancers and may represent a fundamental biological property of basal-like and claudin-low breast cancers. Kaplan-Meier analysis of relapse-free survival revealed a survival disadvantage for patients with breast cancers that exhibit aberrant DNA hypermethylation. Identification of this distinguishing trait among triple-negative breast cancers forms the basis for development of new rational therapies that target the epigenome in patients with basal-like and claudin-low breast cancers.

Illusory movements prevent cortical disruption caused by immobilization.

Enforced limb disuse strongly disrupts the cortical networks that are involved in sensorimotor activities. This disruption causes a cortical reorganization that may be functionally maladaptive. In this study, we used functional magnetic resonance imaging (fMRI) to investigate whether it is possible to prevent this reorganization by compensating for the lack of actual kinesthetic perception with illusory movements induced by "neuromimetic" proprio-tactile feedback that is artificially delivered during immobilization. Sixteen healthy volunteers were equipped for five days with full-hand ortheses that prevented them from performing finger and hand movements but allowed for kinesthetic and tactile sensations. Eight participants received a twice-daily proprio-tactile treatment consisting of the perception of kinesthetic sensations resembling those felt during actual movements generated by miniature vibrators set in the ortheses at the finger and wrist levels. Eight untreated participants received no stimulation. The effects of hand immobilization and treatment were assessed by fMRI during a calibrated voluntary hand movement task and hand tactile stimulation before cast placement and immediately after cast removal. We found that the sensorimotor network was preserved in subjects who underwent this treatment during hand immobilization, while the sensorimotor network of untreated subjects was significantly altered. These findings suggest that sensory feedback and associated movement perception may counteract disuse-induced cortical plastic changes through recruitment of a large part of the cortical network used for actual performed movement. The possibility of guiding cortical plasticity with proprioceptive augmented feedback is potentially relevant for rehabilitation efforts.

ALK-activating homologous mutations in LTK induce cellular transformation.

Leukocyte tyrosine kinase (LTK) is a receptor tyrosine kinase reported to be overexpressed in human leukemia. Though much regarding the function of LTK remains unknown, it shares a high degree of similarity with anaplastic lymphoma kinase (ALK), which is found mutated in human cancer. In order to determine if LTK has transforming potential, we created two LTK mutants, F568L and R669Q, that correspond to two well-characterized activating mutations of ALK (F1174L and R1275Q). LTK-F568L, but not wildtype LTK or LTK-R669Q, transformed hematopoietic cells to cytokine independence. LTK-F568L exhibited a stronger ability to induce loss of contact inhibition and anchorage-independent growth of epithelial cells compared to LTK-R669Q, while wildtype LTK was non-transforming in the same cells. Likewise, LTK-F568L induced greater neurite outgrowth of PC12 cells than R669Q, while wildtype LTK could not. Correlating with transforming activity, LTK-F568L displayed significantly enhanced tyrosine phosphorylation compared to wildtype LTK and LTK-R668Q and induced activation of various signaling proteins including Shc, ERK and the JAK/STAT pathway. Expression of wildtype LTK or LTK-R669Q generally led to weaker activation of signaling proteins than expression of LTK-F568L, or no activation at all. Thus, mutating LTK at residue F568, and to a lesser extent at R669, activates the receptor tyrosine kinase, inducing cell signaling that results in transforming properties. These studies suggest that aberrant activation of LTK may contribute to neoplastic cell growth.

SU-E-T-199: The Radiological Physics Center's Credentialing Dosimetry Reviews: Their Effect on Clinical Trial Deviation Rates.

PURPOSE: To describe the Radiological Physics Center's (RPC) methods to evaluate an institution's ability to meet protocol guidelines in order to decrease NCI clinical trial deviation rate. METHODS: The RPC's dosimetry group utilizes 3 methods of assessing an institutions ability to meet the protocol treatment specifications. These methods involve a clinical and dosimetric review of a treatment plan submitted by the institution prior to the first patient being treated on a protocol. The three evaluation methods include use of site/treatment modality specific benchmark cases, evaluation of a previous patient treated in a similar fashion and a rapid review of the first patient placed on a trial prior to start of treatment. The dosimetric review consists of an independent dose recalculation using RPC measured data or RPC standard dosimetry data. The clinical review assesses the patient's DVHs and contouring of the tumor volume and critical structures, typically in conjunction with a radiation oncologist. RESULTS: Over the past 5 years the RPC has performed these QA reviews for several of the clinical trial groups for several different disease sites and treatment modalities. We have reviewed 1366 treatment plans as a part of credentialing (97 gynecological, 223 prostate, 1046 breast) where 222 failed the first submission requiring the RPC to interact with the submitting institution to resolve the discrepancy. The review of the benchmarks has resulted in 18% of the institutions requiring intervention by the RPC. Performing these reviews has identified potential clinical and dosimetric problem areas that could possibly have resulted in 17% of the charts reviewed to receive a minor or major deviation. CONCLUSIONS: The RPC's clinical and dosimetry review of submitted treatment plans before or early in the treatment process has helped to reduce the deviation rates on protocols. Work supported by PHS grant CA 10953 awarded by NCI, DHHS.

Mutations in the transmembrane and juxtamembrane domains enhance IL27R transforming activity.

Cytokines and their receptors regulate haemopoiesis by controlling cellular growth, survival and differentiation. Thus it is not surprising that mutations of cytokine receptors contribute to the formation of haemopoietic disorders, including cancer. We recently identified transforming properties of IL27R, the ligand-binding component of the receptor for interleukin-27. Although wild-type IL27R exhibits transforming properties in haemopoietic cells, in the present study we set out to determine if the transforming activity of IL27R could be enhanced by mutation. We identified three mutations of IL27R that enhance its transforming activity. One of these mutations is a phenylalanine to cysteine mutation at residue 523 (F523C) in the transmembrane domain of the receptor. The two other mutations identified involve deletions of amino acids in the cytoplasmic juxtamembrane region of the receptor. Expression of each of these mutant IL27R proteins led to rapid cytokine-independent transformation in haemopoietic cells. Moreover, the rate of transformation induced by these mutants was significantly greater than that induced by wild-type IL27R. Expression of these IL27R mutants also induced enhanced activation of JAK (Janus kinase)/STAT (signal transducer and activator of transcription) signalling compared with wild-type. An activating deletion mutation of IL27R enhanced homodimerization of the receptor by a mechanism that may involve disulfide bonding. These transforming IL27R mutants displayed equal or greater transforming activity than bona fide haemopoietic oncogenes such as BCR-ABL (breakpoint cluster region-Abelson murine leukaemia viral oncogene homologue) and JAK2-V617F. Since IL27R is expressed on haemopoietic stem cells, lymphoid cells and myeloid cells, including acute myeloid leukaemia blast cells, mutation of this receptor has the potential to contribute to a variety of haemopoietic neoplasms.

CRLF2 and JAK2 in B-progenitor acute lymphoblastic leukemia: a novel association in oncogenesis.

Expression of cytokine receptor-like factor 2 (CRLF2) has recently been shown to be upregulated as well as mutated in populations of B-progenitor acute lymphoblastic leukemia (B-ALL), including Down syndrome (DS-ALL) patients, lacking recurring chromosomal translocations. Increased CRLF2 expression associates with JAK2 mutation, a combination that transforms hematopoietic cells, suggesting that mutant JAK2 and CRLF2 may cooperate to contribute to B-ALL formation. Importantly, elevated CRLF2 expression correlates with poor outcome in high-risk B-ALL patients. Therefore, CRLF2 may provide a new prognostic marker for high-risk B-ALL, and inhibition of CRLF2/JAK2 signaling may represent a therapeutic approach for this population of ALL patients.

DNMT3b overexpression contributes to a hypermethylator phenotype in human breast cancer cell lines.

BACKGROUND: DNA hypermethylation events and other epimutations occur in many neoplasms, producing gene expression changes that contribute to neoplastic transformation, tumorigenesis, and tumor behavior. Some human cancers exhibit a hypermethylator phenotype, characterized by concurrent DNA methylation-dependent silencing of multiple genes. To determine if a hypermethylation defect occurs in breast cancer, the expression profile and promoter methylation status of methylation-sensitive genes were evaluated among breast cancer cell lines. RESULTS: The relationship between gene expression (assessed by RT-PCR and quantitative real-time PCR), promoter methylation (assessed by methylation-specific PCR, bisulfite sequencing, and 5-aza-2'deoxycytidine treatment), and the DNA methyltransferase machinery (total DNMT activity and expression of DNMT1, DNMT3a, and DNMT3b proteins) were examined in 12 breast cancer cell lines. Unsupervised cluster analysis of the expression of 64 methylation-sensitive genes revealed two groups of cell lines that possess distinct methylation signatures: (i) hypermethylator cell lines, and (ii) low-frequency methylator cell lines. The hypermethylator cell lines are characterized by high rates of concurrent methylation of six genes (CDH1, CEACAM6, CST6, ESR1, LCN2, SCNN1A), whereas the low-frequency methylator cell lines do not methylate these genes. Hypermethylator cell lines coordinately overexpress total DNMT activity and DNMT3b protein levels compared to normal breast epithelial cells. In contrast, most low-frequency methylator cell lines possess DNMT activity and protein levels that are indistinguishable from normal. Microarray data mining identified a strong cluster of primary breast tumors that express the hypermethylation signature defined by CDH1, CEACAM6, CST6, ESR1, LCN2, and SCNN1A. This subset of breast cancers represents 18/88 (20%) tumors in the dataset analyzed, and 100% of these tumors were classified as basal-like, suggesting that the hypermethylator defect cosegregates with poor prognosis breast cancers. CONCLUSION: These observations combine to strongly suggest that: (a) a subset of breast cancer cell lines express a hypermethylator phenotype, (b) the hypermethylation defect in these breast cancer cell lines is related to aberrant overexpression of DNMT activity, (c) overexpression of DNMT3b protein significantly contributes to the elevated DNMT activity observed in tumor cells expressing this phenotype, and (d) the six-gene hypermethylator signature characterized in breast cancer cell lines defines a distinct cluster of primary basal-like breast cancers.

Proprio-tactile integration for kinesthetic perception: an fMRI study.

This study aims to identify the cerebral networks involved in the integrative processing of somesthetic inputs for kinesthetic purposes. In particular, we investigated how muscle proprioceptive and tactile messages can result in a unified percept of one's own body movements. We stimulated either separately or conjointly these two sensory channels in order to evoke kinesthetic illusions of a clockwise rotation of 10 subjects' right hand in an fMRI environment. Results first show that, whether induced by a tactile or a proprioceptive stimulation, the kinesthetic illusion was accompanied by the activation of a very similar cerebral network including cortical and subcortical sensorimotor areas, which are also classically found in passive or imagined movement tasks. In addition, the strongest kinesthetic illusions occurred under the congruent proprio-tactile co-stimulation condition. They were specifically associated to brain area activations distinct from those evidenced under the unimodal stimulations: the inferior parietal lobule, the superior temporal sulcus, the insula-claustrum region, and the cerebellum. These findings support the hypothesis that heteromodal areas may subserve multisensory integrative mechanisms at cortical and subcortical levels. They also suggest the integrative processing might consist of detection of the spatial coherence between the two kinesthetic messages involving the inferior parietal lobule activity and of a detection of their temporal coincidence via a subcortical relay, the insula structure, usually linked to the relative synchrony of different stimuli. Finally, the involvement of the superior temporal sulcus in the feeling of biological movement and that of the cerebellum in the movement timing control are also discussed.

Vibration-induced post-effects: a means to improve postural asymmetry in lower leg amputees?

Muscle vibration has been shown to induce long-lasting and oriented alteration of standing posture in healthy individuals. The postural alterations can last several minutes following the end of vibration and are called post-effects. The goal of this study was to determine whether persons with lower leg amputation that show persistent postural asymmetry after usual rehabilitation experience these postural post-effects and if this could improve their weight bearing on the prosthesis. Centre of pressure (CP) position during stance was recorded prior to and up to 13 min after a 30s unilateral vibration applied during sitting to lateral neck (trapezius) or hip (gluteus medius) muscles in 14 individuals with unilateral lower leg amputation and 18 controls. The amputees' postural asymmetry was confirmed prior to the vibration intervention. A CP displacement, without an increase in CP velocity, was observed in both groups of participants over the 13 min post-vibration. For both the neck or hip vibration sites, the CP shifts were directed in the medio-lateral plane and were oriented either towards the vibrated side or the opposite side across subjects. This led to a decrease of postural asymmetry in half of the group of amputees. Within subject, the orientation of the post-effect was constant and changed to the opposite direction with vibration of the opposite body side. It is suggested that the post-effects are produced by a change of the postural reference consequent to the sustained proprioceptive message induced during the muscle vibration period. The orientation of the post-effects is discussed in relation to the notion of reference frame preference. All in all, because post-effect orientation is constant within subject and adaptive, future studies should investigate if individuals with lower leg amputation could benefit from postural post-effects induced by muscle vibration to improve function.

Cerebral correlates of the "Kohnstamm phenomenon": an fMRI study.

This paper addresses the issue of the central correlates of the "Kohnstamm phenomenon", i.e. the long-lasting involuntary muscle contraction which develops after a prolonged isometric voluntary contraction. Although this phenomenon was described as early as 1915, the mechanisms underlying these post-effects are not yet understood. It was therefore proposed to investigate whether specific brain areas may be involved in the motor post-effects induced by either wrist muscle contraction or vibration using the fMRI method. For this purpose, experiments were carried out on the right wrist of 11 healthy subjects. Muscle activity (EMG) and regional cerebral blood flow were recorded during isometric voluntary muscle contraction and muscle vibration, as well as during the subsequent involuntary contractions (the post-effects) which occurred under both conditions. Brain activations were found to occur during the post-contraction and post-vibration periods, which were very similar under both conditions. Brain activation involved motor-related areas usually responsible for voluntary motor command (primary sensory and motor cortices, premotor cortex, anterior and posterior cingulate gyrus) and sensorimotor integration structures such as the posterior parietal cortex. Comparisons between the patterns of brain activation associated with the involuntary post-effects and those accompanying voluntary contraction showed that cerebellar vermis was activated during the post-effect periods whereas the supplementary motor area was activated only during the induction periods. Although post-effects originate from asymmetric proprioceptive inputs, they might also involve a central network where the motor and somatosensory areas and the cerebellum play a key role. In functional terms, they might result from the adaptive recalibration of the postural reference frame altered by the sustained proprioceptive inputs elicited by muscle contraction and vibration.

Spontaneous bursting neuronal discharges recorded from peripheral nerve in human: injury discharges or not?

This paper deals with a spontaneous, bursting neuronal activity which can not be altered by any stimulation in the periphery or voluntary actions or by cognitive tasks. An initial description of such units led to the conclusion that this activity was generated ectopically at the site of a previous or present impalement of a nerve fibre. The aim of the current study was to record a larger number of these units by using microneurography, in order to characterise their firing properties and particularly, see if any subtypes of units could be identified. In conclusion, this paper suggests that some of these discharges could be related to an injury of the nerve fibre, however most of them could not. Some hypothesis regarding the nature of these bursting activities are suggested.

Long-lasting body leanings following neck muscle isometric contractions.

Our objective was to investigate the neural mechanisms and the functional relevance of motor effects that develop involuntarily following the release of a sustained isometric muscle contraction. The few data available in the literature deal only with post-contractions occurring in a body segment. Although these data emphasise the role of proprioceptive input, the question as to whether this phenomenon is of central or peripheral origin remains unclear. Given the leading role of neck muscle proprioceptive input in body orientation and posture regulation, we designed two experiments to test for postural posteffects after voluntary and involuntary neck muscle contraction. The spatiotemporal characteristics of the posteffects were analysed by means of stabilometric recordings following 30-s isometric contraction of splenius, trapezius and levator muscle groups, and 30-s electrically-induced contraction of the levator muscle group. Results show that a postural response occurred after voluntary contraction of each muscle group tested, which was oriented in the plane of action of the muscle, and lasted 14 min at least. In contrast, no clearly oriented body leanings were found after electrical stimulation of the levator muscle, except for a slight increase in natural postural instability. Data are interpreted as a change in the postural reference resulting from an increase in proprioceptive inflow accompanying mainly the voluntary muscle contraction.

A behavioral approach to understanding and treating pathological gambling.

This report provides a behavioral account of gambling and its treatment. It describes the similarities between gambling and other behaviors maintained by intermittent schedules of reinforcement, the relationship between response cost and gambling behaviors, and how magnitudes of reinforcers affect gambling behaviors. In addition, the relationship between immediacy of reinforcement and behavior is described. Using these behavioral phenomena, behavioral and cognitive-behavioral treatments of pathological gambling are described. Finally, we present the rationale and framework for our cognitive-behavioral treatment, and we provide a behavioral interpretation of 12-step groups.

Levodopa-induced drowsiness in healthy volunteers: results of a choice reaction time test combined with a subjective evaluation of sedation.

The aim of the present study was to assess levodopa (L-Dopa)-induced drowsiness in healthy volunteers using two parameters: choice reaction time and a subjective rating of sedation. Sixteen subjects participated in a randomized, double-blinded, crossover study. A single dose of 200 mg L-Dopa or placebo was administered at 9:00 AM. To limit peripheral side effects connected with L-Dopa, subjects were treated with 20 mg domperidone three times daily. Subjective rating of sedation consisted of visual analogue scale. Reaction time was measured by means of responses to two light-emitting diodes. The illumination of one of these diodes constituted the imperative signal. Manual responses were performed on two buttons located under the right and left index fingers. Results demonstrated a positive correlation between sedation level and reaction time (r = 0.70, p = 0.0026). Adverse events of L-Dopa were nausea (four cases) and excitation (one case). Subjects who did not develop adverse events were faster under L-Dopa than under placebo (p = 0.02), whereas subjects who had nausea or excitation were slower. A single dose of L-Dopa either deteriorated or improved choice reaction time in healthy volunteers according to whether it was sedative and whether it generated disruptive adverse events.

Foot sole and ankle muscle inputs contribute jointly to human erect posture regulation.

In order to assess the relative contribution and the interactions of the plantar cutaneous and muscle proprioceptive feedback in controlling human erect posture, single or combined vibratory stimuli were applied to the forefoot areas and to the tendons of the tibialis anterior muscles of nine standing subjects using various vibration frequency patterns (ranging from 20 to 80 Hz). The variations in the centre of foot pressure, ankle angle and the EMG activities of the soleus and tibialis anterior muscles of each subject were recorded and analysed. Separate stimulation of the plantar forefoot zones or the tibialis anterior muscles always resulted in whole-body tilts oppositely directed backwards and forwards, respectively, the amplitude of which was proportional to the vibration frequency. EMG activity of ankle muscles also varied according to the direction of the postural responses. However, the same vibration frequency did not elicit equivalent postural responses: in the low frequency range, tactile stimulation induced stronger postural effects than proprioceptive stimulation, and the converse was the case for the higher frequency range. Under sensory conflict conditions, i.e. foot sole-flexor ankle muscle co-stimulation, the direction of the body tilts also varied according to the difference and the absolute levels of the vibration frequencies. In all cases, the resulting postural shifts always corresponded to the theoretical sum of the isolated effects observed upon vibrating each of these two sensory channels. We proposed that tactile and proprioceptive information from the foot soles and flexor ankle muscles might be co-processed following a vector addition mode to subserve the maintenance of erect stance in a complementary way.

Impulsivity and voucher versus money preference in polydrug-dependent participants enrolled in a contingency-management-based substance abuse treatment program.

Thirty-four polydrug-dependent participants enrolled in a voucher-based substance abuse treatment program were given choices between hypothetical amounts of money and hypothetical amounts of vouchers, which are traded for goods and services, to determine their preferences for the two payment modalities. It was hypothesized that the majority of participants would prefer money to voucher because under the circumstances of the treatment program, the delay associated with money exchange is shorter than the delay associated with voucher exchange. It was further hypothesized that those participants who selected money over voucher also would have greater levels of impulsivity as assessed by the Barratt Impulsiveness Rating Scale (BIS) (Barratt, 1965). The results show large individual differences in money/voucher preference with approximately half of the participants preferring money to voucher when the two amounts are equivalent. In addition, as the magnitude of the money/voucher comparisons increased from 0.50 dollars to 32.00 dollars, the percentage of participants that preferred money increased. No correlations were found between money/voucher preference and impulsivity scores.

Relations between the directions of vibration-induced kinesthetic illusions and the pattern of activation of antagonist muscles.

In humans, tendon vibration evokes illusory sensations of movement that are usually associated with an excitatory tonic response in muscles antagonistic to those vibrated (antagonist vibratory response, AVR), i.e., in the muscle groups normally contracted if the illusory movement had been performed. The aim of the present study was to investigate the relation between the parameters of the illusory sensation of movement and those of the AVR and to determine whether vectorial models could account for the integration of proprioceptive inputs from several muscles, as well as for the organization of the elementary motor commands leading to one unified motor response. For that purpose, we analyzed the relations between the anatomical site of the tendon vibration, the direction of the illusory movement, the muscles in which the AVR develops, and the characteristics of the AVR (surface EMG, motor unit types, firing rates, and activation latencies). This study confirmed the close relationship between the parameters of an AVR and those of the kinesthetic illusion. It showed that, during illusions of movements in different directions, motor units are activated according to a specific pattern correlated with their type, with the direction of the illusory movement and with the biomechanical properties of their bearing muscles. Finally, kinesthetic illusions and AVRs can be effectively represented using similar vectorial computations. These strong relations between the perceptual and motor effects of tendon vibration once again suggest that the AVR may result from a perceptual-to-motor transformation of proprioceptive information, rather than from spinal reflex mechanisms.

Proprioceptive population coding of two-dimensional limb movements in humans: I. Muscle spindle feedback during spatially oriented movements.

The proprioceptive coding of multidirectional ankle joint movements was investigated, focusing in particular on the question as to how accurately the direction of a movement is encoded when all the proprioceptive information from all the muscles involved in the actual movement is taken into account. During ankle movements imposed on human subjects, the activity of 30 muscle spindle afferents originating in the extensor digitorum longus, tibialis anterior, extensor hallucis longus and peroneus lateralis muscles was recorded from the lateral peroneal nerve using the microneurographic technique. In the first part of the study, it was proposed to investigate whether muscle spindle afferents have a preferred direction, as previously found to occur in the case of cortical cells, and to analyze the neural coding of the movement trajectories using a "population vector model." This model is based on the idea that neuronal coding can be analyzed in terms of a series of vectors, each based on specific movement parameters. In the present case, each vector gives the mean contribution of a population of muscle spindle afferents within one directionally tuned muscle. A given population vector points in the "preferred sensory direction" of the muscle to which it corresponds, and its length is the mean frequency of all the afferents within that muscle. Our working hypothesis was that the sum of these weighted vectors points in the same direction as the ongoing movement. The results show that each muscle spindle afferent, and likewise each muscle, has a specific preferred sensory direction, as well as a preferred sensory sector within which it is capable of sending sensory information to the central nervous system. Interestingly, the results also demonstrate that the preferred directions are the same as the directions of vibration-induced illusions. In addition, the results show that the neuronal population vector model describes the multipopulation proprioceptive coding of spatially oriented 2D limb movements, even at the peripheral sensory level, based on the sum vectors calculated from all the muscles involved in the movement. In an accompanying paper, the coding of more complex 2D movements such as those involved in drawing rectilinear and curvilinear geometrical shapes was investigated.

Proprioceptive population coding of two-dimensional limb movements in humans: II. Muscle-spindle feedback during "drawing-like" movements.

It was proposed to study the proprioceptive sensory coding of movement trajectories during the performance of two-dimensional "drawing-like" movements imposed on the tip of the foot. For this purpose, the activity of the muscle-spindle afferents from the Extensor digitorum longus, Tibialis anterior, Extensor hallucis longus, and Peroneus lateralis muscles was recorded from the lateral peroneal nerve using the microneurographic technique. The drawing movements, describing geometrical shapes such as squares, triangles, ellipses, and circles, were imposed at a constant velocity in both the clockwise and counterclockwise directions. A total number of 44 muscle-spindle afferents were tested, 36 of which were identified as primary and eight as secondary afferents. Whatever the shape of the imposed foot movement, the primary endings from one muscle never discharged throughout the whole trajectory (on average, they discharged for only 49.2% of the length of the trajectory), whereas all the secondary endings discharged for most part of the drawing trajectories (average: 84.8%). The relationship between afferent discharge rate and direction could be described with a cosine-shaped tuning function. The peak of this function corresponded to the preferred sensory direction of the receptor-bearing muscles. The whole path of a given geometrical drawing movement was found to be coded in turn by each of the primary afferents originating from each of the muscles successively stretched. The contribution of each population of muscle afferents from each ankle muscle was represented by a "population vector", whose orientation was the preferred direction of the muscle under consideration and whose length was the mean instantaneous frequency of the afferent population. The "sum vector" corresponding to the sum of all these weighted "population vectors" was found to point in the instantaneous direction of the drawing trajectory, i.e., the tangent to the trajectory. These findings suggest that trajectory information is already encoded at the peripheral level on the basis of the integrated inputs provided by sets of receptors belonging to all the muscles acting on a given joint.