Collection and quality of rhesus monkey semen

Electroejaculation is an accepted method of semen collection from nonhuman primates. Although both penile and rectal probe stimulation techniques have been used, there has been a general lack of consistency and detail regarding their application. This report describes the collection, processing, and evaluation of rhesus monkey semen contrasting two methods of penile electroejaculation: 1) a constant-voltage method where stimulus current is a variable and 2) a constant-current method where stimulus current is operator-controlled. The constant-current method was the more efficient procedure, requiring a lower stimulus current for successful electroejaculation. The influence on semen quality of potentially toxic agents used in the procedure, surgical glove powder and electrolyte cream, was tested; both were detrimental as measured by motility loss. No correlation was found between coagula volume and sperm numbers. The intra- and interanimal variability in semen samples from six monkeys was also evaluated. Penile electroejaculation, combined with control of stimulus current, provides a consistent, successful, and humane method for the collection of semen in the rhesus monkey.     

Role of stimulus-stimulus pairing in matching-to-sample procedure: cross-species comparison of humans and pigeons

The present experiment examined whether, in a matching-to-sample (MTS) procedure, a relation between two stimuli, a sample and a comparison, could be established as a result of just stimulus–stimulus pairing, even if back up reinforcers were never provided for the conditional relation between the sample and comparison stimuli, but rather only for the comparison stimulus. A procedure called “pseudo matching-to-sample” was used in which, when S1 was presented as a sample stimulus, two comparison stimuli (C1 and C2) were presented, and only responses to C1 were reinforced. Conversely, when S2 was presented, only responses to C3 (and not C4) were reinforced. In other words, organisms experiencing this procedure could discriminate C1 from C2, and C3 from C4, by simple discrimination without regard to the conditional sample stimuli. In order to examine cross-species differences, responding by humans in this procedure was compared to that by pigeons. Although the humans developed a discriminative function for the sample stimuli, that is, the humans’ responding was affected by both the sample stimuli and the reinforcers, responding by the pigeons was affected solely by the reinforcers. These data suggest that, in this procedure, humans (but not pigeons) are able to learn relations among stimuli simply as a result of stimulus–stimulus pairing.     

Representation of Faces in Longtailed Macaques (Macaca fascicularis)

The experiments described in this study were intended to increase our knowledge about social cognition in primates. Longtailed macaques (Macaca fascicularis) had to discriminate facial drawings of different emotional expressions. A new experimental approach was used. During the experimental sessions social interactions within the group were permitted, but the learning behaviour of individual monkeys was analysed. The procedure consisted of a simultaneous discrimination between four visual patterns under continuous reinforcement. It has implications not only for simple tasks of stimulus discrimination but also for complex problems of internal representations and visual communication. The monkeys learned quickly to discriminate faces of different emotional expressions. This discrimination ability was completely invariant with variations of colour, brightness, size, and rotation. Rotated and inverted faces were recognized perfectly. A preference test for particular features resulted in a graded estimation of particular facial components. Most important for face recognition was the outline, followed by the eye region and the mouth. An asymmetry in recognition of the left and right halves of the face was found. Further tests involving jumbled faces indicated that not only the presence of distinct facial cues but the specific relation of facial features is essential in recognizing faces. The experiment generally confirms that causal mechanisms of social cognition in non-human primates can be studied experimentally. The behavioural results are highly consistent with findings from neurophysiology and research with human subjects.     

Sensing without touching: psychophysical performance based on cortical microstimulation

Unequivocal proof that the activity of a localized cortical neuronal population provides sufficient basis for a specific cognitive function has rarely been obtained. We looked for such proof in monkeys trained to discriminate between two mechanical flutter stimuli applied sequentially to the fingertips. Microelectrodes were inserted into clusters of quickly adapting (QA) neurons of the primary somatosensory cortex (S1), and the first or both stimuli were then substituted with trains of current pulses during the discrimination task. Psychophysical performance with artificial stimulus frequencies was almost identical to that measured with the natural stimulus frequencies. Our results indicate that microstimulation can be used to elicit a memorizable and discriminable analog range of percepts, and shows that activation of the QA circuit of S1 is sufficient to initiate all subsequent neural processes associated with flutter discrimination.     

Haptic discrimination of size and texture in squirrel monkeys (Saimiri sciureus)

The present study analyzed haptic abilities of four squirrel monkeys. Using a two-alternative forced-choice procedure, stimuli were presented in a visually opaque box, allowing unrestrained test subjects to grab through a small opening and touch the discriminanda. Difference thresholds were determined by a modified method of limits. In the first experiment we determined size difference thresholds for the discrimination of circular cylinders using standard stimuli differing in diameter from 10 mm to 35 mm. In the second experiment a texture difference threshold was obtained for the discrimination of grooved surfaces (groove width 2-7 mm). The squirrel monkeys achieved a mean size difference threshold of 8% stimulus difference. The linear increase of absolute thresholds as a function of the starting stimulus size showed that haptic size discriminations in squirrel monkeys correspond to Weber's law. Three of the animals achieved a texture difference of 10% stimulus difference, while one monkey showed a distinctively lower haptic acuity. An analysis of the exploratory behavior points to a subject-related difference in the significance of cutaneous and kinesthetic information during size discriminations. Whereas differences in the animals' exploratory behavior did not correlate with the size difference threshold a subject achieved, different thresholds for texture discrimination can be explained by the different exploratory procedures the monkeys used to touch grooved surfaces. The low difference thresholds determined for the squirrel monkeys in the present study point to the significance of unrestrained test conditions for the assessment of the haptic capacity of a species.     

Microsecond rotational motions of eosin-labeled myosin measured by time-resolved anisotropy of absorption and phosphorescence

We have studied submicrosecond and microsecond rotational motions within the contractile protein myosin by observing the time-resolved anisotropy of both absorption and emission from the long-lived triplet state of eosin-5-iodoacetamide covalently bound to a specific site on the myosin head. These results, reporting anisotropy data up to 50 microseconds after excitation, extend by two orders of magnitude the time range of data on time-resolved site-specific probe motion in myosin. Optical and enzymatic analyses of the labeled myosin and its chymotryptic digests show that more than 95% of the probe is specifically attached to sulfhydryl-1 (SH1) on the myosin head. In a solution of labeled subfragment-1 (S-1) at 4 degrees C, absorption anisotropy at 0.1 microseconds after a laser pulse is about 0.27. This anisotropy decays exponentially with a rotational correlation time of 210 ns, in good agreement with the theoretical prediction for end-over-end tumbling of S-1, and with times determined previously by fluorescence and electron paramagnetic resonance. In aqueous glycerol solutions, this correlation time is proportional to viscosity/temperature in the microsecond time range. Furthermore, binding to actin greatly restricts probe motion. Thus the bound eosin is a reliable probe of myosin-head rotational motion in the submicrosecond and microsecond time ranges. Our submicrosecond data for myosin monomers (correlation time 400 ns) also agree with previous results using other techniques, but we also detect a previously unresolvable slower decay component (correlation time 2.6 microseconds), indicating that the faster motions are restricted in amplitude. This restriction is not consistent with the commonly accepted free-swivel model of S-1 attachment in myosin. In synthetic thick filaments of myosin, both fast (700 ns) and slow (5 microseconds) components of anisotropy decay are observed. In contrast to the data for monomers, the anisotropy of filaments has a substantial residual component (26% of the initial anisotropy) that does not decay to zero even at times as long as 50 microseconds, implying significant restriction in overall rotational amplitude. This result is consistent with motion restricted to a cone half-angle of about 50 degrees. The combined results are consistent with a model in which myosin has two principal sites of segmental flexibility, one giving rise to submicrosecond motions (possibly corresponding to the junction between S-1 and S-2) and the other giving rise to microsecond motions (possibly corresponding to the junction between S-2 and light meromyosin).(ABSTRACT TRUNCATED AT 400 WORDS)     

Discrete-trial vs. continuous free-operant procedures in assessing whether reinforcement context affects reinforcement value

Pigeons were trained to respond to two alternating concurrent reinforcement schedules. The reinforcement probabilities were .05 and .10 in one component, and .10 and .20 in the other. In one condition, the pigeons received training on a discrete-trial procedure in which the keylights remained illuminated for 5s or until a response occurred. In another condition, pigeons received training on a procedure in which the reinforcement contingencies were the same as in the discrete-trial procedure, but the stimuli were not turned off after 5s or after a response. Following training in each condition, probe tests were presented. In both conditions, the .20 alternative was, overall, preferred to the .05 alternative during probe tests. Following discrete-trial training, there was no reliable preference between the two .10 alternatives. However, when the stimuli remained illuminated during the intertrial interval periods during training, probe tests results showed preference for the .10 alternative that had been presented in the leaner context during training. The pattern of results is consistent with the notion that probe preference can be influenced both by the absolute reinforcement schedules associated with each alternative, as well as changeover behavior developed during training.     

The effect of intruded events on peak time: the role of reinforcement history during the intruded event

Pigeons were studied in an extension of a study by Aum et al. [Aum, S., Brown, B.L., Hemmes, N.S. 2004. The effects of concurrent task and gap events on peak time in the peak procedure. Behav. Process. 65, 43-56] on timing behavior under a discrete-trial fixed-interval (FI) procedure during which 6-s intruded events were superimposed on peak-interval (PI) test trials. In Aum et al., one event consisted in termination of the timing cue (gap trial); the other was a stimulus in the presence of which subjects had been trained to respond under an independent random-interval (RI) schedule of reinforcement (concurrent task trial). Aum et al. found a disruption of timing on concurrent task trials that was greater than that on gap trials. The present study investigated history of reinforcement associated with intruded events as a possible explanation of this earlier finding. After training to peck a side key on a 30-s PI procedure, discrimination training was conducted on the center key in separate sessions; red or green 6-s stimuli were associated with RI 24s or EXT (extinction) schedules. During testing under the PI procedure, three types of intruded events were presented during probe trials--the stimulus associated with the RI (S+) or EXT (S-) schedule during discrimination training, or a gap (termination of the side-keylight). Intruded events occurred 3, 9, or 15s after PI trial onset. Effects of reinforcement history were revealed as substantial disruption of timing during the S+ event and relatively little disruption during the S- event. Intermediate effects were found for the gap event. Results indicate that postcue effects are at least partially responsible for the disruptive effects of the S+ event.     

Water jet: a simple method for classical conditioning in fish

Classical conditioning in animals is a learning procedure involving a biologically relevant stimulus paired with a previously neutral stimulus. In fish, light and sound are frequently used as previously neutral stimuli for conditioning tests. However, in laboratory experiments with replicates, such stimuli may influence the responses of fish in nearby aquariums. Herein, we developed a simple applicable methodology for classical conditioning in fish that prevents this type of influence. We isolated fish in individual aquariums and introduced a water jet that caused localized water movement, followed by the introduction of a food pellet. These procedures were repeated for each fish for 20 days. After 14 days, all fish were conditioned. Moreover, in subsequent probe trials (memory retention tests) conducted within 32 days after conditioning procedures, fish responded accordingly. These findings corroborate the applicability and usefulness of the method tested herein especially under lab conditions. Therefore, we suggest that a simple water jet is a useful and reliable tool for fish conditioning in future studies.     

Amount of training and stimulus salience affect associability changes in serial conditioning

Q. J. Exp. Psychol. 44B (1992) 17 reported that extended training with a serial compound of two conditioned stimuli, S1-->S2, resulted in losses in the rate at which S1-food associations were formed in a subsequent test phase. However, this ability of S1 to participate in new learning (its associability) could be restored by omitting S2 on some trials. Variables that affect performance in this task were examined in five Pavlovian appetitive conditioning experiments with rats. In Experiment 1, the associability of S1 decreased gradually as a function of the number of training sessions with a consistent S1-S2 relation. In Experiment 2, after extended exposure to a consistent S1-S2 relation, the associability of S1 was restored with as few as one training session in which the S1-S2 relation was made inconsistent by omitting S2 on some trials. In Experiments 3-5, test phase conditioning of S1 after inconsistent training was found to be either greater than, less than, or similar to, conditioning of S1 after consistent training, depending on the relative salience of the events used as S1 and S2. The results showed that both intrinsic and extrinsic relations among stimuli must be considered when interpreting the results of experiments in stimulus selection.     

Response-reinforcer relations and resistance to change

Behavioral momentum theory suggests that the relation between a response and a reinforcer (i.e., response-reinforcer relation) governs response rates and the relation between a stimulus and a reinforcer (i.e., stimulus-reinforcer relation) governs resistance to change. The present experiments compared the effects degrading response-reinforcer relations with response-independent or delayed reinforcers on resistance to change in conditions with equal stimulus-reinforcer relations. In Experiment 1, pigeons responded on equal variable-interval schedules of immediate reinforcement in three components of a multiple schedule. Additional response-independent reinforcers were available in one component and additional delayed reinforcers were available in another component. The results showed that resistance to disruption was greater in the components with added reinforcers than without them (i.e., better stimulus-reinforcer relations), but did not differ for the components with added response-independent and delayed reinforcement. In Experiment 2, a component presenting immediate reinforcement alternated with either a component that arranged equal rates of reinforcement with a proportion of those reinforcers being response independent or a component with a proportion of the reinforcers being delayed. Results showed that resistance to disruption tended to be either similar across components or slightly lower when response-reinforcer relations were degraded with either response-independent or delayed reinforcers. These findings suggest that degrading response-reinforcer relations can impact resistance to change, but that impact does not depend on the specific method and is small relative to the effects of the stimulus-reinforcer relation.     

Discrete-trial control of morphine self-injection behaviour in monkeys: effects of injection dose and trials per session.

Responding for intravenous injections of morphine was studied using a discrete-trials procedure in squirrel monkeys. For one group, each session consisted of 10 trials at an inter-trial interval of 15 min; for the second group, each session consisted of 100 trials at an inter-trial interval of 1.5 min. When different injection doses of morphine (1-1000 mug/kg per injection), including saline as a control procedure, were substituted at random for blocks of five consecutive sessions, the frequency of morphine self-administration was found to be an inverted U-shaped function of the injection dose. This relationship was observed in each group of monkeys, despite a 10-fold difference in the total amount of the drug which was available for self-administration per session when a given injection dose was substituted for both groups. The results show that the injection dose of morphine acted as a primary determinant of response probability, even under circumstances in which trial spacing imposed a significant delay between consecutive opportunities for drug self-administration,     

Short-term item memory in successive same-different discriminations

Pigeons were tested in a successive same-different (S/D) discrimination procedure to examine the short-term memory for individual items in sequences of different or identical pictures. Item-by-item analyses of pecking behavior within single trials revealed this S/D discrimination emerged at the earliest possible point in the sequence--the presentation of the second item. Further, by comparing peck rates at points where different types of sequences diverged (e.g. ABA versus ABC), we determined that the pigeons remembered the first item for at least 4-8s and across one to two intervening items. These results indicate that this S/D discrimination was controlled by relational comparisons of pictorial content across memories of specific items, rather than the detection of low-level perceptual "transients" between items. A second experiment supported this conclusion by showing increased discrimination with longer first item viewing times, consistent with encoding of details about individual pictures. These findings further support a qualitative similarity among birds and primates in possessing a general capacity to judge certain types of stimulus relations, such as stimulus identity and difference. Implications for the temporal continuity of experience in animals are also considered.     

Effect of signaling reinforcement on resistance to change in a multiple schedule

This study evaluated the effect of a signal on resistance to change using a multiple schedule of reinforcement. Experiment 1 presented pigeons with three schedules: a signaled delay to reinforcement schedule (a two-link chain schedule with a variable-interval 120-s initial link followed by a 5-s fixed-time schedule), an unsignaled delay schedule (a comparable two-link tandem schedule), and an immediate, zero-delay variable-interval 125-s schedule. Two separate disruption procedures assessed resistance to change: extinction and adding a variable-time 20-s schedule of reinforcement to the inter-component interval. Resistance to change tests were conducted twice, once with the signal stimulus (the terminal link of the chain schedule) present and once with it absent. Results from both disruption procedures showed that signal absence reduced resistance to change for the pre-signal stimulus. In probe choice tests subjects strongly preferred the signal stimulus over the unsignaled stimulus and exhibited no reliable preference when given a choice between the signal stimulus and immediate stimulus. Experiment 2 presented two equal signaled schedules where, during resistance to change tests, the signal remained for one schedule and was removed for the second. Resistance to change was consistently lower when the signal was absent.     

When and how well can human‐socialized capuchins match actions demonstrated by a familiar human?

Capuchin monkeys have provided uneven evidence of matching actions they observe others perform. In accord with theories emphasizing the attentional salience of object movement and spatial relationships, we predicted that human-reared monkeys would better match events in which a human demonstrator moved an object into a new relation with another object or surface than other kinds of actions. Three human-reared capuchins were invited repeatedly by a familiar human to perform a fixed set of actions upon objects or upon their bodies, using the "Do as I do" procedure. Actions directed at the body were matched less reliably than actions involving objects, and actions were matched best when the monkey looked at the demonstration for at least 2 sec and performed its action within a few seconds after the demonstration. The most commonly matched actions were those that one monkey performed relatively often when the experiment began. One monkey partially reproduced three novel actions (out of 48 demonstrations), all three involving moving or placing objects, and two of which it also performed following other demonstrations. These findings contribute convergent evidence that capuchin monkeys display social facilitation of activity, enhanced interest in particular objects and emulation of spatial outcomes. This pattern can support the development of shared manipulative skills, as evident in traditions of foraging and tool use in natural settings. The findings do not suggest that human rearing substantively altered capuchins' ability or interest in matching the actions of a familiar human, although visual attention to the human demonstrator may have been greater in these monkeys than in normally reared monkeys.     

Optimization of a fluorescent-based phosphor imaging dot blot DNA hybridization assay to assess E. coli virulence gene profiles

To increase the efficiency and consistency in screening Escherichia coli for virulence genes, a Phosphor Imager was adopted for signal detection in Dot Blot DNA hybridization replacing X-ray film read by eye. We assessed not only the reliability of the instrument-based procedure, but the impact of going from an outcome measured by visualization on a semi-quantitative scale to a digitized readout on an interval scale. We analyzed technical and biological variability of the assay and the factors contributing to the variability. In spite of high variability both within and between membranes in the Phosphor Imager readings, we were able to define classification rules for gene presence that were remarkably consistent. Using the X-ray film signal detection procedure with Southern confirmation as a gold standard, we obtained a sensitivity and specificity of 87-99% for a rule requiring no retesting for all but one gene probe.     

Noninvasive technique for the repeated sampling of salivary free cortisol in awake,unrestrained squirrel monkeys

The use of noninvasive measures of hypothalamic-pituitary-adrenal (HPA) axis function is of growing interest among preclinical and clinical investigators. This report describes a method for the repeated assessment of salivary free cortisol in awake, unrestrained squirrel monkeys (Saimiri sciureus) based on a saliva sampling technique previously developed for rhesus monkeys. Individually housed adult male squirrel monkeys were trained to chew on dental rope attached to a pole, from which saliva was extracted by centrifugation and analyzed for cortisol by radioimmunoassay (RIA). Eight of nine monkeys readily acquired the task, reliably providing adequate saliva samples for the assay. Salivary free cortisol levels were examined in these subjects under basal conditions and in response to two types of neuroendocrine challenge. Levels of salivary free cortisol showed relatively low intra- and interindividual variability, with mean individual morning levels ranging between 17.1 and 37.9 microg/dl. Squirrel monkeys demonstrated a consistent daily rhythm in salivary free cortisol ranging from a high of 27.4 +/- 5.2 microg/dl (mean +/- SEM) at 12 P.M. to a low of 7.5 +/- 1.6 microg/dl at 6 P.M. Intravenous (IV) challenges with 1 microg/kg ACTH, or 10 and 50 microg/kg CRF resulted in significant increases in salivary free cortisol. The described sampling technique provides a reliable and sensitive means for repeated measurement of HPA activity in unrestrained, awake squirrel monkeys. In addition, our findings illustrate several features of HPA system rhythmicity and reactivity using salivary cortisol instead of blood plasma or serum.     

Variability of Prefrontal Neuronal Discharges before and after Training in a Working Memory Task

Variability of neural discharges can be revealing about the computations and network properties of neuronal populations during the performance of cognitive tasks. We sought to quantify neuronal variability in the prefrontal cortex of naïve monkeys that were only required to fixate, and to examine how this measure was altered by learning and execution of a working memory task. We therefore performed analysis of a large database of recordings in the same animals, using the same stimuli, before and after training. Our results indicate that the Fano Factor, a measure of variability, differs across neurons depending on their functional properties both before and after learning. Fano Factor generally decreased after learning the task. Variability was modulated by task events and displayed lowest values during the stimulus presentation. Nonetheless, the decrease in variability after training was present even prior to the presentation of any stimuli, in the fixation period. The greatest decreases were observed comparing populations of neurons that exhibited elevated firing rate during the trial events. Our results offer insights on how properties of the prefrontal network are affected by performance of a cognitive task.     

灵长动物初级概括能力的比较实验研究(Ⅱ)金丝猴对刺激物大小关系的概括

人们通常所说的客观事物的大小,是指其相对关系而言。离开客体间的绝对尺寸的比较,也就无所谓大小了。所以没有比较就没有对客体大小属性本身的认识,因而也就不能对它们之间的关系进行概括。格式塔心理家认为在辨别学习中被试是学习了刺激物之间的关系。在特定的试验中,被试不是对绝对刺激值进行反应,而是根据刺激的相对属性,在辨别学习中是如此,在泛化测验中更是如此。因此,被试在这里已经摆脱开具体对象的绝对属性,而将这些属性的关系作为线索,并作出正确的反应。刺激物的绝对属性是可能不断改变的,但它们之间的关系则往往是稳定的,于是被试就能够排除其它因素的干扰而对这种关系作出较稳定的反应。这种关系之所以能同具体的绝对属性具有等值作用,无疑是得力于动物的初级概括能力。西方行为灵长类学家们把灵长动物的这种初级概括叫作“概念的形成”(Brown等,1958; Robinson, 1960; Hicks,1956 ),我们认为这种说法是片面的。     

The effects of concurrent task and gap events on peak time in the peak procedure

The effect of a concurrent task on timing performance of pigeons was investigated with the peak interval procedure. Birds were trained to peck a side key on a discrete-trial schedule that included reinforced fixed-interval (FI) 30-s trials and nonreinforced extended probe trials. Then, in separate sessions, birds were trained to peck a 6-s center key for food. In a subsequent test phase, the FI procedure was in effect along with dual-task probe test trials. On those test trials, the 6-s center key (task cue) was presented at 3, 9, or 15s after probe trial onset. During another test phase, a 6-s gap (the FI keylight was extinguished) was presented at 3, 9, or 15s after probe trial onset. Peak time increased with center key time of onset, and was greater under task than gap conditions. Moreover, peak time under task conditions exceeded values predicted by stop and reset clock mechanisms. These results are at variance with current attentional accounts of timing behavior in dual-task conditions, and suggest a role of nontemporal factors in the control of timing behavior.