Cyclic-ratio Schedule Research Articles

Evaluating the cyclic ratio schedule as an assay of feeding behaviour in the European starling (Sturnus vulgaris).

The cyclic ratio (CR) schedule is a behavioural assay developed to study feeding in rats, in which the number of operant responses required to obtain food reward (the ratio requirement) increases and then decreases in a repeating cycle. In a recent study, we used the CR schedule with European starlings (Sturnus vulgaris) to investigate the effects of an early-life manipulation on adult feeding behaviour. As this was the first time the CR schedule had been used with any bird species, a more in-depth evaluation is warranted. Here, we performed a fuller CR experiment with the same birds as the prior study, a year later. First, we examine the individual consistency of feeding behaviour between experimental sessions and also between CR schedules comprising different ratio requirement progressions. We found that between-session consistency was poor to moderate, and that a geometric ratio progression provided greater between-session consistency than an arithmetic ratio progression. Second, we tried to replicate some of the canonical findings from rats working on CR schedules. In contrast to findings from rats, we found that defence of feeding rates did not increase when starlings were acutely food deprived. However, as in rats, we found that the post-reinforcement pause increased linearly with the upcoming ratio requirement, suggesting that starlings were able to learn the cyclic nature of the schedule. Third, we compared the results from the present study concerning the impacts of our early-life treatment with those from our earlier study. We found that the majority of our previous findings were replicated in the same individuals one year on, reinforcing our previous conclusion that the early-life manipulation had canalised our birds into two groups with different patterns of feeding rate defence.

A preclinical screen to evaluate pharmacotherapies for the treatment of agitation in dementia.

Agitation associated with dementia is frequently reported clinically but has received little attention in preclinical models of dementia. The current study used a 7PA2 CM intracerebroventricular injection model of Alzheimer's disease (AD) to assess acute memory impairment, and a bilateral intrahippocampal (IH) injection model of AD (aggregated Aβ1-42 injections) and a bilateral IH injection model of dementia with Lewy bodies (aggregated NAC61-95 injections) to assess chronic memory impairment in the rat. An alternating-lever cyclic-ratio schedule of operant responding was used for data collection, where incorrect lever perseverations measured executive function (memory) and running response rates (RRR) measured behavioral output (agitation). The results indicate that bilateral IH injections of Aβ1-42 and bilateral IH injections of NAC61-95 decreased memory function and increased RRRs, whereas intracerebroventricular injections of 7PA2 CM decreased memory function but did not increase RRRs. These findings show that using the aggregated peptide IH injection models of dementia to induce chronic neurotoxicity, memory decline was accompanied by elevated behavioral output. This demonstrates that IH peptide injection models of dementia provide a preclinical screen for pharmacological interventions used in the treatment of increased behavioral output (agitation), which also establish detrimental side effects on memory.

Lack of support for bexarotene as a treatment for Alzheimer's disease

Bexarotene has been reported to reduce brain amyloid-β (Aβ) levels and to improve cognitive function in transgenic mouse models of Alzheimer's disease (AD). Four groups failed to fully replicate the primary results but the original authors claimed overall support for the general conclusions. Because of its potential clinical importance, the current work studied the effects of bexarotene using two animal species and highly relevant paradigms. Rats were tested for the ability of bexarotene to prevent changes induced by an Aβ challenge in the form intracerebroventricular (i.c.v) administration of 7PA2 conditioned medium (7PA2 CM) which contains high levels of Aβ species. Bexarotene had no effect on the long-term potentiation of evoked extracellular field excitatory postsynaptic potentials induced by i.c.v. 7PA2 CM. It also had no effect following subcutaneous administration of 2, 5, 10 and 15 mg/kg on behavioral/cognitive impairment using an alternating-lever cyclic-ratio schedule of operant responding in the rat. The effects of bexarotene were further tested using the APPSwFILon, PSEN1*M146L*L286V transgenic mouse model of AD, starting at the time Aβ deposits first begin to develop. Mice were sacrificed after 48 days of exposure to 100 mg bexarotene per day. No significant difference between test and control mice was found using a water-maze test, and no significant difference in the number of Aβ deposits in cerebral cortex, using two different antibodies, was apparent. These results question the potential efficacy of bexarotene for AD treatment, even if instigated in the preclinical period prior to the onset of cognitive deficits reported for human AD.This article is part of the Special Issue entitled ‘Synaptopathy – from Biology to Therapy’.

MRZ-99030 – A novel modulator of Aβ aggregation: II – Reversal of Aβ oligomer-induced deficits in long-term potentiation (LTP) and cognitive performance in rats and mice

β-amyloid1-42 (Aβ1-42) is a major endogenous pathogen underlying the aetiology of Alzheimer's disease (AD). Recent evidence indicates that soluble Aβ oligomers, rather than plaques, are the major cause of synaptic dysfunction and neurodegeneration. Small molecules that suppress Aβ aggregation, reduce oligomer stability or promote off-pathway non-toxic oligomerization represent a promising alternative strategy for neuroprotection in AD. MRZ-99030 was recently identified as a dipeptide that modulates Aβ1-42 aggregation by triggering a non-amyloidogenic aggregation pathway, thereby reducing the amount of intermediate toxic soluble oligomeric Aβ species. The present study evaluated the relevance of these promising results with MRZ-99030 under pathophysiological conditions i.e. against the synaptotoxic effects of Aβ oligomers on hippocampal long term potentiation (LTP) and two different memory tasks. Aβ1-42 interferes with the glutamatergic system and with neuronal Ca2+ signalling and abolishes the induction of LTP. Here we demonstrate that MRZ-99030 (100–500 nM) at a 10:1 stoichiometric excess to Aβ clearly reversed the synaptotoxic effects of Aβ1-42 oligomers on CA1-LTP in murine hippocampal slices. Co-application of MRZ-99030 also prevented the two-fold increase in resting Ca2+ levels in pyramidal neuron dendrites and spines triggered by Aβ1-42 oligomers. In anaesthetized rats, pre-administration of MRZ-99030 (50 mg/kg s.c.) protected against deficits in hippocampal LTP following i.c.v. injection of oligomeric Aβ1-42. Furthermore, similar treatment significantly ameliorated cognitive deficits in an object recognition task and under an alternating lever cyclic ratio schedule after the i.c.v. application of Aβ1-42 and 7PA2 conditioned medium, respectively. Altogether, these results demonstrate the potential therapeutic benefit of MRZ-99030 in AD.

RS-0406 Arrests Amyloid-β Oligomer-Induced Behavioural Deterioration In Vivo

Clinically accessible compounds that arrest or reverse the effects of amyloid-β (Aβ) on progressively developing behavioural symptomatology and neuropathology in Alzheimer's disease (AD) have yet to become available. However, a viable strategy may be to target and neutralise soluble Aβ oligomers, which have been shown to mediate synaptic dysfunction and to produce cognitive deficits in the intact organism. Inhibiting the aggregation of Aβ is therapeutically attractive, as Aβ aggregation is a pathological event and pharmacological interventions targeting this are likely to have a non-toxic profile. A behavioural assay, the alternating-lever cyclic-ratio schedule, was used to assess the effect of Aβ oligomers and the non-peptide small molecule RS-0406 in male Sprague-Dawley rats. RS-0406 has been shown to inhibit Aβ1-42 fibrillogenesis and protect against Aβ1-42–induced cytotoxicity in primary hippocampal neurons. In the current study, RS-0406 ameliorated the adverse effects of secreted oligomers of human Aβ on behaviour and dose dependently reduced the behavioural effects of Aβ oligomers, with the highest dose, 10μM, maintaining behaviour approximately at control levels. This effect appeared to be central; peripheral confounds having been extensively investigated. This is the first published report on the effects of RS-0406 in vivo and indicates that RS-0406 has potential as a pharmacotherapeutic intervention for behavioural deficits seen in the early stages of AD, and possibly as an intervention in the development of AD neuropathology. Indeed, an analogue of RS-0406 that could be administered peripherally might be a realistic candidate for the clinical treatment of AD.

An operant determination of the behavioral mechanism of benzodiazepine enhancement of food intake

A recent review paper by Cooper (Appetite 44:133-150, 2005) has pointed out that a role for benzodiazepines as appetite stimulants has been largely overlooked. Cooper's review cited several studies that suggested the putative mechanism of enhancement of food intake after benzodiazepine administration might involve increasing the perceived pleasantness of food (palatability). The present study examined the behavioral mechanism of increased food intake after benzodiazepine administration. The cyclic-ratio operant schedule has been proposed as a useful behavioral assay for differentiating palatability from regulatory effects on food intake (Ettinger and Staddon, Physiol Behav 29:455-458, 1982 and Behav Neurosci 97:639-653, 1983). The current study employed the cyclic-ratio schedule to determine whether the effects on food intake of chlordiazepoxide (CDP) (5.0 mg/kg), sodium pentobarbital (5.0 mg/kg), and picrotoxin (1.0 mg/kg) were mediated through palatability or regulatory processes. The results of this study show that both the benzodiazepine CDP and the barbiturate sodium pentobarbital increased food intake in a manner similar to increasing the palatability of the ingestant, and picrotoxin decreased food intake in a manner similar to decreasing the palatability of the ingestant. These results suggest that the food intake enhancement properties of benzodiazepines are mediated through a mechanism affecting perceived palatability.

Behavioral and neurochemical mechanisms of the action of mild stress in the enhancement of feeding.

Rats were trained to respond under a cyclic-ratio schedule of reinforcement composed of an ascending, followed by a descending, sequence of ratio values. Subjects were trained while exposed to 70 dB white noise, then tested while exposed to 70 or 90 dB white noise. Exposure to 90 dB white noise elevated the response function (p<.02). Naloxone was then administered intraperitoneally at 0.3. 1.0. and 3.0 mg/kg under 70 dB and 90 dB white noise. Naloxone administration (1.0 and 3.0 mg/kg) significantly depressed the response function obtained under 90 dB white noise (ps<.01) but did not affect the function obtained under 70 dB white noise. These findings suggest that mild stress increases food intake through a mechanism affecting palatability enhanced by the release of endogenous opioids.

Effects of reinforcement magnitude and ratio values on behaviour maintained by a cyclic ratio schedule of reinforcement.

In Experiments 1 and 2, lever pressing by rats was reinforced on a cyclic ratio schedule of food reinforcement, comprising a repeated sequence of fixed-ratio component schedules. Reinforcement magnitude was varied, on occasional sessions in Experiment 1 and across blocks of sessions in Experiment 2, from one to two or three 45-mg food pellets. In the one-pellet condition, post-reinforcement pauses increased with component schedule value. At higher magnitudes, post-reinforcement pauses increased, and overall response rates declined. Response rate on component schedules was a decreasing linear function of the obtained rate of reinforcement in all conditions. Plotted against component schedule value, response rate increased exponentially to an asymptote that decreased when reinforcement magnitude increased. These findings are consistent with regulatory accounts of food-reinforced behaviour. In Experiment 3, rats were trained under a cyclic ratio schedule comprising fixed-ratio components including higher values, and some inverted U-shaped response functions were obtained. Those rats that did not show this relationship were trained on cyclic ratios with even higher values, and all showed inverted U-shaped response functions. This suggests that behaviour on cyclic ratio schedules can reflect activating of reinforcement as well as the satiating effects seen in Experiments 1 and 2.

Effects of reinforcement magnitude and ratio values on behaviour maintained by a cyclic ratio schedule of reinforcement

In Experiments 1 and 2, lever pressing by rats was reinforced on a cyclic ratio schedule of food reinforcement, comprising a repeated sequence of fixed-ratio component schedules. Reinforcement magnitude was varied, on occasional sessions in Experiment 1 and across blocks of sessions in Experiment 2, from one to two or three 45-mg food pellets. In the one-pellet condition, post-reinforcement pauses increased with component schedule value. At higher magnitudes, post-reinforcement pauses increased, and overall response rates declined. Response rate on component schedules was a decreasing linear function of the obtained rate of reinforcement in all conditions. Plotted against component schedule value, response rate increased exponentially to an asymptote that decreased when reinforcement magnitude increased. These findings are consistent with regulatory accounts of food reinforced behaviour. In Experiment 3, rats were trained under a cyclic ratio schedule comprising fixed-ratio components including higher values, and some inverted U-shaped response functions were obtained. Those rats that did not showthis relationship were trained on cyclic ratios with even higher values, and all showed inverted U-shaped response functions. This suggests that behaviour on cyclic ratio schedules can reflect activating of reinforcement as well as the satiating effects seen in Experiments 1 and 2.

Alternating lever cyclic-ratio schedule analysis of the effects of atropine sulfate

Cholinergic dysfunction has been implicated in the behavioral and memory impairment that is the hallmark of conditions such as delirium and Alzheimer's Disease. Anticholinergic drugs have been widely used in procedures designed to mimic aspects of the pathology of these conditions in rats. Procedures in use vary widely in sensitivity and behavioral specificity and may be confounded by administration of high drug doses that may not be physiologically relevant. The current study proposes the use of rats responding on an alternating lever cyclic-ratio schedule to study the effects of the anticholinergic compound atropine sulfate. This procedure enables simultaneous measurement of anticipatory ratio tracking (postreinforcement pause durations), perseverations (lever switching errors), and nonspecific peripheral drug effects (running response rates). Results of this study suggest that the schedule is sensitive to low drug doses (0.1–1.0 mg/kg atropine), measures the ability to track changing ratio conditions and to execute lever alternation, and allows for monitoring of peripheral drug effects during behavioral testing. The procedure's sensitivity and low effective dose range may make it useful in the study of behaviors related to anticholinergic effects.

Cyclic-ratio schedule analysis of a serotonin agonist and depletor on consummatory behaviour

Four male Wistar rats, 180 days old at the start of the experiment, at 85% of their free-feeding body weight were trained to respond on a geometric cyclic-ratio schedule comprised of the following ratio values: 2, 4, 8, 16, 32 and 64, for 0.1 ml of 5% sucrose reinforcement. The response functions (response rates plotted against reinforcement rates) were linear and of negative slope over the range of ratio values from 2 to 16. IP administration of 1.0 mg/kg 5-MeODMT reduced the x- and y-intercepts of the linear portion of the response function without altering the slope relative to 2.0 ml/kg 0.85% saline IP. This was interpreted as a perceived palatability effect. IP administration of 100 mg/kg pCPA elevated the reinforcement rate intercept but also decreased the slope of the response function. This finding was interpreted as an increase in the perceived palatability of the reinforcer, coupled with a decrease in motivation at higher schedule conditions, possibly due to peripheral effects of pCPA.

Cholecystokinin, diet palatability, and feeding regulation in rats

Rats ate less food than normal on cyclic-ratio schedules following cholecystokinin and lithium chloride injections. Nevertheless, they defended this lower eating rate in the same way as under control conditions. The pattern of effects produced by cholecystokinin and lithium chloride resembled those following diet adulteration with citric acid and sucrose octa acetate and differed from the effects produced by increases in body weight. Cholecystokinin and lithium chloride injections also produced similar changes in the free-feeding patterns of non-deprived rats: Both meal size and intermeal intervals decreased in a manner similar to the effects of citric acid and sucrose octa acetate adulteration. Interpreted in terms of a static regulatory model, these results suggest that cholecystokinin and lithium chloride suppress feeding by degrading the palatability of food, not by promoting satiety, discomfort, or illness.

Operant regulation of feeding: a static analysis.

Cyclic-ratio schedules are a rapid method for studying the operant regulation of feeding rate. The cyclic method produces results comparable to traditional but time-consuming parametric methods. Performance on cyclic-ratio schedules is well described by a linear regulatory model that embodies three quantitative feedback assumptions: (a) that rate of feeding is regulated by the rate of operant behavior, (b) that taste factors have an additive effect on the rate of the operant response, and (c) that regulatory "gain" is inversely related to body weight. This model accurately describes poorer regulatory performance at high body weights and following amphetamine administration, and the effects of altered diet palatability on preferred feeding rates.

Decreased feeding associated with acute hypoxia in rats

Rats obtained less food than normal on a cyclic-ratio schedule during brief, 1-hr exposure to either moderate hypobaric hypoxia (BP=435 Torr, PO 2≈91 Torr) or to hypoxic hypoxia (BP=750 Torr, PO 2≈90 Torr), but not during hypobaric exposure with 36.5% oxygen (BP=435 Torr, PO 2≈159 Torr). The depressed rate of feeding associated with hypoxia was nevertheless well regulated. Interpreted in terms of a regulatory model, these results suggest that hypoxia suppresses eating because it degrades the taste of food, not because it impairs feeding regulation or general activity.