Making the connection: How membrane contact sites have changed our view of organelle biology.

The view of organelles and how they operate together has changed dramatically over the last two decades. The textbook view of organelles was that they operated largely independently and were connected by vesicular trafficking and the diffusion of signals through the cytoplasm. We now know that all organelles make functional close contacts with one another, often called membrane contact sites. The study of these sites has moved to center stage in cell biology as it has become clear that they play critical roles in healthy and developing cells and during cell stress and disease states. Contact sites have important roles in intracellular signaling, lipid metabolism, motor-protein-mediated membrane dynamics, organelle division, and organelle biogenesis. Here, we summarize the major conceptual changes that have occurred in cell biology as we have come to appreciate how contact sites integrate the activities of organelles.

Form follows function: the importance of endoplasmic reticulum shape.

The endoplasmic reticulum (ER) has a remarkably complex structure, composed of a single bilayer that forms the nuclear envelope, along with a network of sheets and dynamic tubules. Our understanding of the biological significance of the complex architecture of the ER has improved dramatically in the last few years. The identification of proteins and forces required for maintaining ER shape, as well as more advanced imaging techniques, has allowed the relationship between ER shape and function to come into focus. These studies have also revealed unexpected new functions of the ER and novel ER domains regulating alterations in ER dynamics. The importance of ER structure has become evident as recent research has identified diseases linked to mutations in ER-shaping proteins. In this review, we discuss what is known about the maintenance of ER architecture, the relationship between ER structure and function, and diseases associated with defects in ER structure.

The influence of shark liver oils on normal and transformed mammalian cells in culture.

n3 polyunsaturated fatty acids have been reported to have anti-carcinogenic effects on mammalian carcinomas. n3 fatty acids occur in high concentrations in marine oils, especially shark liver oils. Several reports have indicated an extremely low incidence of cancer in sharks, although other reports indicate carcinogenesis in some shark species. It has been hypothesised that n3 fatty acids and other components of shark liver oil may exert anti-carcinogenic effects. The aim of this study was to assess whether shark liver oil, from four Indian Ocean shark species, exerted anti-proliferative effects on transformed and normal mammalian cells in culture, and to assess whether the ratio of n3 to n6 polyunsaturates influenced the results. Neither the shark liver oils themselves, nor the ratio of n3 to n6, showed any consistently significant effects with either transformed or normal cells.

Uptake and trafficking of exogenous sterols in Saccharomyces cerevisiae.

The proper distribution of sterols among organelles is critical for numerous cellular functions. How sterols are sorted and moved among membranes remains poorly understood, but they are transported not only in vesicles but also by non-vesicular pathways. One of these pathways moves exogenous sterols from the plasma membrane to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae. We have found that two classes of proteins play critical roles in this transport, ABC transporters (ATP-binding-cassette transporters) and oxysterol-binding protein-related proteins. Transport is also regulated by phosphoinositides and the interactions of sterols with other lipids. Here, we summarize these findings and speculate on the role of non-vesicular sterol transfer in determining intracellular sterol distribution and membrane function.

Self-induced versus reactive triggering of synchronous movements in a deafferented patient and control subjects.

The present study investigates the contribution of tactile-kinesthetic information to the timing of movements. The relative timing of simultaneous tapping movements of finger and foot (hand-foot asynchrony) was examined in a simple reaction time task and in discrete self-initiated taps (Experiment 1), and in externally triggered synchronization tapping (Experiment 2). We compared the performance of a deafferented participant (IW) to the performance of two control groups of different ages. The pattern of results in control groups replicates previous findings: Whereas positive hand-foot asynchronies (hand precedes foot) are observed in a simultaneous reaction to an auditory stimulus, hand-foot asynchronies are negative with discrete self-initiated as well as auditorily paced sequences of synchronized finger and foot taps. In the first case, results are explained by a simultaneous triggering of motor commands. In contrast, self-initiated and auditorily paced movements are assumed to be controlled in terms of their afferent consequences, as provided by tactile-kinesthetic information. The performance of the deafferented participant differed from that of healthy participants in some aspects. As expected on the basis of unaffected motor functions, the participant was able to generate finger and foot movements in reaction to an external signal. In spite of the lack of movement-contingent sensory feedback, the deafferented participant showed comparable timing errors in self-initiated and regularly paced tapping as observed in control participants. However, in discrete self-initiated taps IW's hand-foot asynchronies were considerably larger than in control participants, while performance did not differ from that of controls in continuous movement generation. These findings are discussed in terms of an internal generation of the movement's sensory consequences (forward-modeling).

Perceptual basis of bimanual coordination.

Periodic bimanual movements are often the focus of studies of the basic organizational principles of human actions. In such movements there is a typical spontaneous tendency towards mirror symmetry. Even involuntary slips from asymmetrical movement patterns into symmetry occur, but not vice versa. Traditionally, this phenomenon has been interpreted as a tendency towards co-activation of homologous muscles, probably originating in motoric neuronal structures. Here we provide evidence contrary to this widespread assumption. We show for two prominent experimental models-bimanual finger oscillation and bimanual four-finger tapping-that the symmetry bias is actually towards spatial, perceptual symmetry, without regard to the muscles involved. We suggest that spontaneous coordination phenomena of this kind are purely perceptual in nature. In the case of a bimanual circling model, our findings reveal that highly complex, even 'impossible' movements can easily be performed with only simple visual feedback. A 'motoric' representation of the performed perceptual oscillation patterns is not necessary. Thus there is no need to translate such a 'motoric' into a 'perceptual' representation or vice versa, using 'internal models' (ref. 29). We suggest that voluntary movements are organized by way of a representation of the perceptual goals, whereas the corresponding motor activity, of sometimes high complexity, is spontaneously and flexibly tuned in.

Interactions between perception and action in a reaction task with overlapping S-R assignments.

We have used a novel task to study relationships between perception and action. Four experiments studied stimulus-response (S-R) relationships under conditions in which stimuli and responses were functionally unrelated (i.e., not assigned to each other by instruction) and merely overlapped in time. On each trial, participants carried out movements on a graphic tablet while observing motions displayed on a computer screen. The movement on trial n was specified by the motion observed on the previous trial n-1, whereas the motion observed on trial n specified the movement to be performed on trial n + 1. Results showed that stimulus motion had a contrast-like impact on response movement. Watching a small motion while performing a medium-sized movement increased movement size, whereas watching a large motion led to a decrease (Experiment 1). Further experiments showed that the contrast pattern was not affected by the mode of motion presentation (Experiment 2), or by the interval between motion and movement execution (Experiment 3). Contrast was also observed in the reverse direction, i.e., from action to perception (Experiment 4). We propose that the contrast effect is due to a mechanism for selective code modification. This mechanism acts to increase the distinctiveness of simultaneously activated perception and action codes in a common representational domain.

Motor learning enhances perceptual judgment: a case for action-perception transfer.

Transfer from perception to action is well documented, for instance in the form of observational learning. Transfer from action to perception, on the other hand, has not been researched. Such action-perception transfer (APT) is compatible with several learning theories and has been predicted within the framework of common coding of perceptual and motor events (Prinz, 1992, 1997). Our first experiment aimed at an empirical evaluation of APT and involved motor practice of timed two-cycle arm movements on verbal command without visual feedback. In a transfer test, visual judgments of similar patterns had to be made. In addition, transfer from the visual to the motor task was studied. In Experiment 2 we separated kinesthetic aspects of motor practice from preparatory and efferent contributions to APT. The experiments provide evidence that transfer between perception and action is bi-directional. Transfer from perception to action and, more importantly, from action to perception was found. Furthermore, APT was equally pronounced for participants who had actively practiced movements during training and for passive participants who had received merely kinesthetic feedback about the movement. This kinesthetic-visual transfer is likely to be achieved via visuomotor-kinesthetic matching or via timekeeping mechanisms that are involved in both motor and visual performance.

Recognition of self-generated actions from kinematic displays of drawing.

Five experiments addressed the question of whether individuals can distinguish between self-generated and other-generated actions when seeing their visual effects. Each experiment consisted of a recording session in which participants drew familiar and unfamiliar characters without receiving visual feedback and a recognition session in which they provided self-or-other judgments (SOJs) to indicate whether a kinematic display reproduced the visual effects of their own actions. The main results were that self-generated and other-generated drawing can be distinguished, that the familiarity of character shapes does not influence the accuracy of SOJs, and that velocity information is crucial for the identification of self-generated drawing. The ability to determine authorship from kinematic displays of drawing provides evidence for the contribution of action-planning structures to perception.

Tapping with peripheral nerve block. a role for tactile feedback in the timing of movements.

This study examines the impact of peripheral nerve block, that is, the elimination of tactile feedback on synchronization performance. In a tapping experiment in which subjects were instructed to tap in synchrony with an auditory pacing signal, three different tasks were studied under conditions with and without peripheral nerve block: standard tapping with tactile contact, isometric tapping, and contact-free tapping. In addition, the maximum tapping rate was registered both with and without peripheral nerve block. It was found that the anticipatory error, usually observed in synchronization tasks, was affected by the peripheral nerve block in the standard tapping and the isometric tapping task. In both tasks, local anesthesia led to an increase in asynchrony between the pacing signal and the tap. Performance remained unimpaired in those tasks in which tactile information was assumed to play a minor role (maximum tapping rate and contact-free tapping). The results clearly demonstrate the importance of tactile feedback for the timing of movements. The predictions of a model assuming a strong correlation between the amount of sensory feedback and the size of the negative asynchrony in synchronization tasks were examined and discussed.

Movement observation affects movement execution in a simple response task.

The present study was designed to examine the hypothesis that stimulus-response arrangements with high ideomotor compatibility lead to substantial compatibility effects even in simple response tasks. In Experiment 1, participants executed pre-instructed finger movements in response to compatible and incompatible finger movements. A pronounced reaction time advantage was found for compatible as compared to incompatible trials. Experiment 2 revealed a much smaller compatibility effect for less ideomotor-compatible object movements compared to finger movements. Experiment 3 presented normal stimuli (hand upright) and flipped stimuli (hand upside-down). Two components were found to contribute to the compatibility effect, a dynamic spatial compatibility component (related to movement directions) and an ideomotor component (related to movement types). The implications of these results for theories about stimulus-response compatibility (SRC) as well as for theories about imitation are discussed.

Directing attention to movement effects enhances learning: a review.

Studies investigating the influence of the learner's focus of attention, induced by instructions or feedback, on motor skill learning are reviewed. In general, directing performers' attention to the effects of their movements (external focus of attention) appears to be more beneficial than directing their attention to their own movements (internal focus of attention). Preliminary evidence is presented indicating that an internal attentional focus constrains the motor system by interfering with natural control processes, whereas an external focus seems to allow automatic control processes to regulate the movements. Support for the view that actions are controlled by their anticipated effects comes from research demonstrating functional variability in motor control, as well as the benefits of purposeful activity in occupational therapy. We explain these results in terms of the ideomotor principle of human actions (James, 1890) and its more modern derivatives (Hommel, 1996; Prinz, 1990, 1997).

An analysis of ideomotor action.

This article presents a framework based on the work of R. H. Lotze (1852), W. James (1890), and A. G. Greenwald (1970) for understanding ideomotor actions that tend to arise when individuals watch others perform certain actions. Two principles of ideomotor action induction are distinguished: perceptual induction, in which people tend to perform the movements they see, and intentional induction, in which people tend to perform movements suited to achieve what they would like to see. In 3 experiments, ideomotor hand, head, and foot movements were studied while participants watched a ball traveling toward a target. Results showed strong support for intentional induction, weaker support for perceptual induction, and a strong impact of the effector studied. The representational basis of action induction (stimulus vs. goal representations) and the automaticity of the underlying processing (task-dependent vs. task-independent induction) were considered.

The Theory of Event Coding (TEC): a framework for perception and action planning.

Traditional approaches to human information processing tend to deal with perception and action planning in isolation, so that an adequate account of the perception-action interface is still missing. On the perceptual side, the dominant cognitive view largely underestimates, and thus fails to account for, the impact of action-related processes on both the processing of perceptual information and on perceptual learning. On the action side, most approaches conceive of action planning as a mere continuation of stimulus processing, thus failing to account for the goal-directedness of even the simplest reaction in an experimental task. We propose a new framework for a more adequate theoretical treatment of perception and action planning, in which perceptual contents and action plans are coded in a common representational medium by feature codes with distal reference. Perceived events (perceptions) and to-be-produced events (actions) are equally represented by integrated, task-tuned networks of feature codes--cognitive structures we call event codes. We give an overview of evidence from a wide variety of empirical domains, such as spatial stimulus-response compatibility, sensorimotor synchronization, and ideomotor action, showing that our main assumptions are well supported by the data.

Correspondence effects with manual gestures and postures: a study of imitation.

In this study, the authors applied methods and theories from research of stimulus-response compatibility (SRC) to action imitation. In 6 experiments, they adopted the logic of the Simon paradigm (B. Hommel & W. Prinz, 1996) to explore interference between task-relevant symbolic stimulus features (color) and task-irrelevant iconic stimulus features (2 hand gestures and 2 postures). The same 2 hand gestures served as responses. Pronounced correspondence effects for both gestures and postures showed up throughout. In line with theories of SRC, the authors account for these correspondence effects in terms of overlap arising between stimulus and response features in a common representational domain. As a specific extension of this approach, they propose 2 functionally independent mechanisms: One operates movement-based when dynamic information is provided, and the other operates state-based with static postures as stimuli. Implications for theories of both SRC and action imitation are discussed.

Launching the effect: representations of causal movements are influenced by what they lead to.

We investigated whether the representation of an observed causal movement is influenced by its observed effect. Subjects watched displays showing collisions between two objects. In this "launching event" (Michotte, 1946/1963), one of the two objects (Object A) started to move and set a second, initially stationary, object (Object B) into motion, which gave a strong impression of apparent causality. The apparent effectiveness of A's movement was manipulated by varying the velocities of A and B. When the velocity of B was higher than that of A, the effectiveness of the collision was high; when it was smaller it was low. Then, subjects were asked to reproduce the velocity of the causal movement. Reproduced velocity followed the velocity of both Object A and Object B, which supports the hypothesis that the effect of a movement is integrated with its apparent cause. However, when apparent causality was reduced by changing the direction of motion of B or by covering the point of collision, the influence of the effect on the representation of the cause persisted, suggesting that retroactive interference may account for the findings. The interference effect could not be reduced to temporal recency or spatial integration and was not obtained in the reverse temporal order (proactive interference). Rather, the two successive movements were blended in memory.

Compatibility between observed and executed finger movements: comparing symbolic, spatial, and imitative cues.

Intuitively, one can assume that imitating a movement is an easier task than responding to a symbolic stimulus like a verbal instruction. Support for this suggestion can be found in neuropsychological research as well as in research on stimulus-response compatibility. However controlled experimental evidence for this assumption is still lacking. We used a stimulus-response compatibility paradigm to test the assumption. In a series of experiments, it was tested whether observed finger movements have a stronger influence on finger movement execution than a symbolic or spatial cue. In the first experiment, we compared symbolic cues with observed finger movements using an interference paradigm. Observing finger movements strongly influenced movement execution, irrespective of whether the finger movement was the relevant or the irrelevant stimulus dimension. In the second experiment, effects of observed finger movements and spatial finger cues were compared. The observed finger movement dominated the spatial finger cue. A reduction in the similarity of observed and executed action in the third experiment led to a decrease of the influence of observed finger movement, which demonstrates the crucial role of the imitative relation of observed and executed action for the described effects. The results are discussed in relation to recent models of stimulus-response compatibility. Neurocognitive support for the strong relationship between movement observation and movement execution is reported.

Neuromagnetic correlates of sensorimotor synchronization.

Sensorimotor synchronization tasks, in which subjects have to tap their finger in synchrony with an isochronous auditory click, typically reveal a synchronization error with the tap preceding the click by about 20 to 50 msec. Although extensive behavioral studies and a number of different explanatory accounts have located the cause of this so-called "negative asynchrony" on different levels of processing, the underlying mechanisms are still not completely understood. Almost nothing is known about the central processes, in particular, which sensory or motor events are synchronized by subjects. The present study examined central-level processing in synchronization tasks with magnetoencephalography (MEG). Eight subjects synchronized taps with their right index finger to an isochronous binaural pacing signal presented at an interstimulus interval of 800 msec. To gain information on central temporal coupling between "tap" and "click," evoked responses were averaged time-locked to the auditory signal and the tap onset. Tap-related responses could be explained with a three dipole model: One source, peaking at approximately 77 msec before tap onset, was localized in contralateral primary motor cortex (MI); the two other sources, peaking approximately at tap onset and 75 msec after tap onset, in contralateral primary somatosensory cortex (SI). Temporal coupling of these sources was compared in relation to different trigger points. The second SI source was equally well time-locked to the tap and to the auditory click. Furthermore, analysis of the time locking of this source activity as a function of the temporal order of tap and click showed that the second event - irrespective whether tap or click - was decisive in triggering the second SI source. This suggests that subjects use mainly sensory feedback in judging and evaluating whether they are "keeping time."

Mutants affecting the structure of the cortical endoplasmic reticulum in Saccharomyces cerevisiae.

We find that the peripheral ER in Saccharomyces cerevisiae forms a dynamic network of interconnecting membrane tubules throughout the cell cycle, similar to the ER in higher eukaryotes. Maintenance of this network does not require microtubule or actin filaments, but its dynamic behavior is largely dependent on the actin cytoskeleton. We isolated three conditional mutants that disrupt peripheral ER structure. One has a mutation in a component of the COPI coat complex, which is required for vesicle budding. This mutant has a partial defect in ER segregation into daughter cells and disorganized ER in mother cells. A similar phenotype was found in other mutants with defects in vesicular trafficking between ER and Golgi complex, but not in mutants blocked at later steps in the secretory pathway. The other two mutants found in the screen have defects in the signal recognition particle (SRP) receptor. This receptor, along with SRP, targets ribosome-nascent chain complexes to the ER membrane for protein translocation. A conditional mutation in SRP also disrupts ER structure, but other mutants with translocation defects do not. We also demonstrate that, both in wild-type and mutant cells, the ER and mitochondria partially coalign, and that mutations that disrupt ER structure also affect mitochondrial structure. Our data suggest that both trafficking between the ER and Golgi complex and ribosome targeting are important for maintaining ER structure, and that proper ER structure may be required to maintain mitochondrial structure.

Determinants of translocation and folding of TreF, a trehalase of Escherichia coli.

One isoform of trehalase, TreF, is present in the cytoplasm and a second, TreA, in the periplasm. To study the questions of why one enzyme is exported efficiently and the other is not and whether these proteins can fold in their nonnative cellular compartment, we fused the signal sequence of periplasmic TreA to cytoplasmic TreF. Even though this TreF construct was exported efficiently to the periplasm, it was not active. It was insoluble and degraded by the periplasmic serine protease DegP. To determine why TreF was misfolded in the periplasm, we isolated and characterized Tre(+) revertants of periplasmic TreF. To further characterize periplasmic TreF, we used a genetic selection to isolate functional TreA-TreF hybrids, which were analyzed with respect to solubility and function. These data suggested that a domain located between residues 255 and 350 of TreF is sufficient to cause folding problems in the periplasm. In contrast to TreF, periplasmic TreA could fold into the active conformation in its nonnative cellular compartment, the cytoplasm, after removal of its signal sequence.