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Target selection in drug discovery--defined here as the decision to focus on finding an agent with a particular biological action that is anticipated to have therapeutic utility--is influenced by a complex balance of scientific, medical and strategic considerations. In this article, we provide an introduction to the key issues in target selection and discuss the rationale for decision making.
Photoreceptor neurons (R cells) in the Drosophila visual system elaborate a precise map of visual space in the brain. The eye contains some 750 identical modules called ommatidia, each containing eight photoreceptor cells (R1-R8). Cells R1-R6 synapse in the lamina; R7 and R8 extend through the lamina and terminate in the underlying medulla. In a screen for visual behavior mutants, we identified alleles of flamingo (fmi) that disrupt the precise maps elaborated by these neurons. These mutant R1-R6 neurons select spatially inappropriate targets in the lamina. During target selection, Flamingo protein is dynamically expressed in R1-R6 growth cones. Loss of fmi function in R cells also disrupts the local pattern of synaptic terminals in the medulla, and Flamingo is transiently expressed in R8 axons as they enter the target region. We propose that Flamingo-mediated interactions between R-cell growth cones within the target field regulate target selection.
During development of the mammalian cerebral cortex, thalamic axons must grow into the telencephalon and select appropriate cortical targets. In order to begin to understand the cellular interactions that are important in cortical target selection by thalamic axons, we have examined the morphology of axons from the lateral geniculate nucleus (LGN) as they navigate their way to the primary visual cortex. The morphology of geniculocortical axons was revealed by placing the lipophilic tracer Dil into the LGN of paraformaldehyde-fixed brains from fetal and neonatal cats between embryonic day 26 (E26; gestation is 65 d) and postnatal day 7 (P7). This morphological approach has led to three major observations. (1) As LGN axons grow within the intermediate zone of the telencephalon toward future visual cortex (E30-40), many give off distinct interstitial axon collaterals that penetrate the subplate of nonvisual cortical areas. These collaterals are transient and are not seen postnatally. (2) There is a prolonged period during which LGN axons are restricted to the visual subplate prior to their ingrowth into the cortical plate; the first LGN axons arrive within visual subplate by E36 but are not detected in layer 6 of visual cortex until about E50. (3) Within the visual subplate, LGN axons extend widespread terminal branches. This represents a marked change in their morphology from the simple growth cones present earlier as LGN axons navigate en route to visual cortex. The presence of interstitial collaterals suggests that there may be ongoing interactions between LGN axons and subplate neurons along the entire intracortical route traversed by the axons. From the extensive branching of LGN axons within the visual subplate during the waiting period, it appears that they are not simply "waiting." Rather, LGN axons may participate in dynamic cellular interactions within the subplate long before they contact their ultimate target neurons in layer 4. These observations confirm the existence of a prolonged waiting period in the development of thalamocortical connections and provide important morphological evidence in support of the previous suggestion that interactions between thalamic axons and subplate neurons are necessary for cortical target selection.
Retroviral integrase plays an important role in choosing host chromosomal sites for integration of the cDNA copy of the viral genome. The domain responsible for target site selection has been previously mapped to the central core of the protein (amino acid residues 49-238). Chimeric integrases between human immunodeficiency virus type 1 (HIV-1) and feline immunodeficiency virus (FIV) were prepared to examine the involvement of a nonspecific DNA-binding region (residues 213-266) and certain alpha helices within the core domain in target site selection. Determination of the distribution and frequency of integration events of the chimeric integrases narrowed the target site-specifying motif to within residues 49-187 and showed that alpha 3 and alpha 4 helices (residues 123-166) were not involved in target site selection. Furthermore, the chimera with the alpha 2 helix (residues 118-121) of FIV identity displayed characteristic integration events from both HIV-1 and FIV integrases. The results indicate that the alpha 2 helix plays a role in target site preference as either part of a larger or multiple target site-specifying motif.
We recorded the activity of single neurons in the middle temporal (MT) and middle superior temporal (MST) visual areas in two macaque monkeys while the animals performed a smooth pursuit target selection task. The monkeys were presented with two moving stimuli of different colors and were trained to initiate smooth pursuit to the stimulus that matched the color of a previously given cue. We designed these experiments so that we could separate the component of the neuronal response that was driven by the visual stimulus from an extraretinal component that predicted the color or direction of the selected target. We found that for all cells in MT and MST the response was primarily determined by the visual stimulus. However, 14% (8 of 58) of MT neurons and 26% (22 of 84) of MST neurons had a small predictive component that was significant at the P < or = 0.05 level. In some cells, the predictive component was clearly related to the color of the intended target, but more often it was correlated with the direction of the target. We have previously documented a systematic shift in the latency of smooth pursuit that depends on the relative direction of motion of the two stimuli. We found that neither the latency nor the amplitude of neuronal responses in MT or MST was correlated with behavioral latency. These results are consistent with a model for target selection in which a weak selection bias for the intended target is amplified by a competitive network that suppresses motion signals related to the nonintended stimulus. It is possible that the predictive component of neuronal responses in MT and MST contributes to the selection bias. However, the strength of the selection bias in MT and MST is not sufficient to account for the high degree of selectivity shown by pursuit behavior.
Academic researchers are increasingly producing and using cDNA microarrays. Their quality and hybridization specificity are crucial in determining whether the generated data are accurate and interpretable. Here, we describe two methods of monitoring microarray production, the sustainability of DNA attachment, and the specificity of hybridization. The first method consists of labeling an oligonucleotide, which is one of the primers used to amplify all cDNA probes on the array (except for beta-actin and GAPDH) with fluorescent dye and hybridize it to the cDNA microarray. Attachment of the cDNAs on the array after the hybridization procedure was monitored by visualizing fluorescent signals from the spots on the array. In the second method, two selected DNA targets, beta-actin and GAPDH, were labeled with fluorescent dye to hybridize to the cDNA array. Hence, hybridization specificity was demonstrated by obtaining fluorescent signals solely from the genes corresponding to the target.
Attending to a relevant item in a visual display is thought to require not only selective attention to this item, but also active inhibition of surrounding distractor items. As a consequence of this spatial inhibition, selection of a relevant item in a previous distractor location is slowed (i.e., the spatial inhibition effect). The goal of this study is to identify brain regions that are involved in this spatial inhibition effect using functional magnetic resonance imaging (fMRI). Subjects had to select a target from a display which also included a distractor, while that target was presented in either a new location (control) or in a location previously occupied by a distractor (spatial inhibition). A region of interest analysis revealed decreased activation in the superior parietal lobe (SPL), but increased activation in the motor areas (supplementary motor area, putamen) when the target was presented in a previously inhibited compared to a new location. We take these results to suggest that presenting a target in a previously inhibited location negatively biases the selection of that target in favor of an accompanying distractor. This may result in an initially more efficient selection process, resulting in lower activation in the SPL. Counteracting this perceptual bias possibly requires additional motor activation. This study provides evidence for the notion that to make selection more efficient, prior information concerning an item is used. When this prior information conflicts with the current stimulus demands, compensatory motor actions are taken to correct this perceptual bias.
Critical to the discovery and development of drugs and vaccines is the rational selection of biochemical, immunologic or molecular targets. To understand the rationale for target selection, we review strengths and weaknesses of the four main approaches: whole animal disease models; molecular targeting; epidemiology/observation studies, and genomics. After classifying diseases into those with a relatively stable pathophysiology (e.g., hypertension and gout) versus those with an unstable pathophysiology (e.g., AIDS and influenza) to aid in understanding target selection, we provide examples of successful and unsuccessful selection of drug and vaccine targets, focusing on the molecular and epidemiological/observational approaches. We discuss the reasons that molecular targeting has led to successful control of many diseases, whereas the epidemiological/observational approach has had a checkered history. We also assess the potential power of the genomic approach, specifically the curative versus controlling/preventive strategies. With combined genetic and molecular approaches and judicious use of whole animal models and properly performed epidemiology/observation studies to select the appropriate targets, the future for controlling, preventing and even curing many diseases is very bright indeed.
Previous studies have examined the facilitative effects of prior spatial information on target selection for saccadic eye movements. More recently, studies have shown that prior spatial information also influences target selection for smooth pursuit. However, direct comparisons of the effects of prior information on target selection for pursuit and saccades have not been made. To this end, we provided different classes of prior information and measured their effects on target selection for pursuit and saccades. In Experiment 1, we assessed the relative effects of spatial cues (indicating the target stimulus' initial location) and color cues (indicating the target stimulus' color) on eye movement latencies. In Experiment 2, we assessed the effects of motion cues (indicating the target stimulus' direction of motion) in addition to spatial cues. For both pursuit and saccades, we found that spatial cues reduced eye movement latencies more than color cues (Experiment 1). Spatial cues also reduced eye movement latencies more than motion cues (Experiment 2), even for pursuit, despite the fact that stimulus motion is essential for the generation of pursuit eye movements. These results indicate that both pursuit and saccades are affected to a greater degree by spatial information than motion or color information. We suggest that the primacy of spatial information for both pursuit and saccades reflects the importance of spatial attention in selecting the stimulus target for both eye movements.
The purpose of the present study was to selectively target pigmented trabecular meshwork cells without producing collateral damage to adjacent non-pigmented cells or structures. The ability to selectively target trabecular meshwork cells without coagulation, while preserving the structural integrity of the meshwork, could be a useful approach to study whether the biological response of non-coagulative damage to the trabecular meshwork and trabecular meshwork cells is similar to that seen with coagulative damage to the trabecular meshwork which occurs with argon laser trabeculoplasty. This approach also may be useful to non-invasively deplete trabecular meshwork cells while preserving the structural integrity of the trabecular meshwork in an animal model. A mixed cell culture of pigmented and non-pigmented trabecular meshwork cells were irradiated with Q-switched Nd-YAG and frequency-doubled Nd-YAG lasers, microsound pulsed dye-lasers, and an argon ion laser in order to define a regime where laser absorption would be confined to pigmented trabecular meshwork cells, thereby permitting selective targeting of these cells without producing collateral thermal damage to adjacent non-pigmented cells. Pulse durations ranged from 10 nsec to 0.1 sec. A fluorescent viability/cytotoxicity assay was used to evaluate laser effects and threshold energies, and cells were examined morphologically by light and TEM. Selective targeting of pigmented trabecular meshwork cells was achieved with pulse durations between 10 nsec and 1 microsec and 1 microsec without producing collateral thermal or structural damage to adjacent non-pigmented trabecular meshworks cells when examined by light and transmission electron microscopy. Pulse durations greater than 1 microsec resulted in non-selective killing of non-pigmented trabecular meshwork cells. Threshold radiant exposures were as low as 18 mJ cm-2, and increased at longer wavelengths, longer pulse durations and lower melanin contents within the cells. It is concluded that selective targeting of pigmented trabecular meshwork cells can be achieved using pulsed lasers with low threshold radiant exposures avoiding collateral thermal damage to adjacent non-pigmented trabecular meshwork cells. This approach can be readily applied in vivo.
We explored the effects of microstimulation on target selection by delivering stimulation at different depths within V1 (striate cortex) of the rhesus monkey (Macaca mulatta). Stimulation evoked saccadic eye movements that terminated in the receptive-field location of the activated neurons. The current thresholds for saccade evocation were highest (> or = 30 micro A) in the superficial layers and lowest (< or = 10 micro A) in the deep layers. To study target selection, one visual target was presented in the receptive-field location of the stimulated neurons and a second visual target was presented outside this location. Microstimulation delivered in concert with the appearance of the two targets decreased the probability that a monkey would select the target placed in the receptive-field location when the upper layers of V1 were stimulated, and it increased this probability when the lower layers were stimulated. We suggest that microstimulation of the upper layers of V1 disrupts visual signals from retina en route to higher cortical areas, whereas microstimulation of the lower layers activates V1 efferents that innervate the oculomotor system.
The ability of human beings to shift their vergence between multiple disparity stimuli was investigated. The stimulus was a stereogram consisting of a single bar (1 x 0.3 degree) projected in the center of a larger circular pattern (28 degrees dia) of dots (0.3 degree dia). In the initial condition, the subjects perceived a single bar in the center of a cluster of dots, all lying in a single depth plane. The subjects were instructed to fixate either the bar or one of the dots close to the bar. Stepwise changes of target-vergence of either the bar, or of all dots, or of both configurations in opposite directions, were imposed in a random sequence. Under these conditions, ocular vergence was controlled by the target-vergence of the selected target at all times, even if this implied a loss of binocular fusion for the non-selected target. The effect of target selection per se, without concomitant changes in retinal position of the selected target, was studied in two experienced subjects with stabilized image techniques. The subjects viewed a configuration containing short vertical bars placed at either side of a long vertical bar. The short bars on different sides of the long bar had opposite (crossed or uncrossed) disparities. After stabilization of the configuration while the subject fixated the long bar, subjects attempted to fixate the short bars alternatingly. Due to the stabilization the ensuing eye movements did not affect the retinal positions, and thus the disparity, of the bars. Attempted fixation of a parafoveally viewed, short bar induced vergence responses in the appropriate direction.(ABSTRACT TRUNCATED AT 250 WORDS)
Microstimulation of the intermediate layers of V1 in rhesus monkeys disrupts target selection with saccadic eye movements. To study target selection, one visual target was presented in the receptive-field location of the stimulated neurons and a second target was presented outside this location. Microstimulation delivered with the appearance of the two targets decreased the probability that the monkey would select the target placed in the receptive-field location when intermediate layers of V1 were stimulated. This interference effect was more pronounced when anodal-first pulse pairs were used as compared to when cathodal-first pulse pairs were used. The superior effectiveness of anodal pulses suggests that the interference effect is due to activation of axonal terminals residing within intermediate V1.
Do deeper levels of processing produce equivalent priming effects at all stages of task performance? In Experiment 1, we varied the level of processing factorially across two task stages--target selection and response selection. Each stage required perceptual (e.g., color) or conceptual (e.g., friendliness) processing of stimulus items (i.e., animal names). Negative priming was substantially greater when deeper processing was required at the target selection stage, but it was unaffected by the level of processing at the response selection stage. In contrast, positive priming was greater when deeper processing was required at the response selection stage, but it was unaffected by processing at the target selection stage. In Experiment 2, we generalized this finding using a task in which numeric targets were selected on the basis of their parity. As in Experiment 1, the deeper level of processing at the target selection stage produced a larger negative priming effect. These results illuminate the role of target selection demands in modulating the strength of negative priming.
Previous studies have shown that a LATER (Linear Approach to Threshold with Ergodic Rate) race model can be used to explain saccadic target selection and latencies. The goal of the present study was to determine whether a comparable model could be applied to the underlying decision-making processes involved in target selection for transient vergence eye movements in rhesus monkeys. Luminance contrast of near and far Gabor pair stimuli were manipulated in a forced-choice paradigm to investigate their influence on vergence target selection. The distributions of responses and their latencies were evaluated by cumulative recinormal and reciprobit plots. With all targets set to 20% luminance contrast, animals showed a bias for the divergent target. Increasing luminance contrast of the near Gabor pair, while holding the far Gabor at the base contrast, resulted in increasing selection of the convergent target. This change in bias from divergent to convergent target selection correlated with decreases in convergent latency and increases in divergent latency. Monte Carlo simulations were used to estimate the internal rates of the divergent and convergent decision-making processes which, given a fixed threshold, would result in the observed distributions of vergence responses and their latencies. Statistical tests show that the LATER race model can predict observed values, and strongly suggests that competition between internal convergent and divergent target selection processes determines relative frequencies and latencies of these movements.
To investigate the development of the target selection process for saccade generation, saccade latencies toward one of two targets were compared with those toward a target presented alone, under conditions where the timing of fixation stimulus offset was varied. 12 2-mo.-olds (7 boys and 5 girls), 12 3-mo.-olds (6 boys and 6 girls), and 12 5-mo.-olds (8 boys and 4 girls) participated. The latencies were longer when two saccade targets were presented on the left and right sides of the display (Double-target condition) than when one target was presented either on the left or right side of the display (Single-target condition). The difference between the Single and Double-target conditions tended to be larger in 2-mo. old infants than in 5-mo.-old infants when a central fixation target remained on after the onset of peripheral stimuli (Overlap condition). However, there was no difference between the 2-mo.-olds and the 5-mo.-olds when the fixation target was turned off before the peripheral stimuli were presented (Gap condition). The results provided evidence of the maturational development of a target selection process for saccade generation during 2 to 5 mo. of age.
Because real-world scenes typically contain many different potential objects of interest, selecting one goal from many is clearly a fundamental problem faced by the saccadic system. We recorded from visual, movement, and visuo-movement (VM) neurons in the superior colliculus (SC) of monkeys performing a reaction-time visual-search task requiring them to make saccades to an odd-colored target presented with distractors. First, we compared the responses of SC neurons in search with their responses when a single target was presented without distractors (single-stimulus task). Consistent with earlier reports, initial visual activity was smaller in search than in the single-stimulus task, while movement-related activity in the two tasks was comparable. Further experiments showed that much of the reduction in the initial visual response during search was due to lateral inhibition, although a top-down task-related component was also evident. Although the initial visual activity did not discriminate the target from the distractors, some neurons showed a biphasic pattern of visual activity. In VM burst neurons, the second phase of this activity was significantly larger when the target, rather than a distractor, was in the response field. We traced the time course of target/distractor discrimination using receiver operating characteristic (ROC) analysis and found that VM burst neurons, VM prelude neurons, and pure movement neurons discriminated the target from distractors before saccade onset but that phasic and tonic pure visual neurons did not. We also examined the relationship between target/distractor discrimination time and saccade latency. Discrimination in VM burst neurons having a biphasic pattern of visual activity and in many VM prelude neurons occurred after a consistent delay that did not depend on saccade latency, suggesting that these neurons are involved in target selection as well as movement initiation. In contrast, VM burst neurons lacking a biphasic pattern of visual activity, pure movement neurons, and a subset of VM prelude neurons discriminated the target at a time that was well correlated with saccade latency, suggesting that this latter group of neurons is involved in triggering movement execution but not in target selection. Thus a mix of signals likely related to target selection and movement initiation co-exists in different groups of SC neurons. This suggests that certain types of SC neurons participate in the target selection process and that the SC as a whole represents a gateway for target selection signals to be converted into a saccadic command.
Saccades are rapid eye movements that orient gaze toward areas of interest in the visual scene. Neural activity correlated with saccade target selection has been identified in several brain regions, including the superior colliculus (SC), but it is not known whether the SC is directly involved in target selection, or whether the SC merely receives selection-related signals from cortex in preparation for the execution of eye movements. In monkeys, we used focal reversible inactivation to test the functional contributions of the SC to target selection during visual search, and found that inactivation resulted in clear deficits. When a target appeared in the inactivated field, saccades were often misdirected to distractor stimuli. Control tasks showed that this deficit was not caused by low-level visual or motor impairments. Our results indicate that, in addition to its well-established involvement in movement execution, the SC has an important functional role in target selection.