Separation of short and medium-chain fatty acids using capillary electrophoresis with indirect photometric detection: Part I: Identification of fatty acids in rat feces.

Short and medium-chain fatty acids (SMCFAs) are known as essential metabolites found in gut microbiota that function as modulators in the development and progression of many inflammatory conditions as well as in the regulation of cell metabolism. Currently, there are few simple and low-cost analytical methods available for the determination of SMCFA. This report focuses on SMCFA analysis utilizing CE with indirect photometric detection (CE-IPD). A ribonucleotide electrolyte, 5'-adenosine mono-phosphate (5'-AMP), is investigated as an IPD reagent due to its high molar absorptivity and dynamic reserve compatible with separation and detection of SMCFA. The operating parameters like the composition of organic solvent, millimolar concentrations of the complexing agent (alpha-cyclodextrin), 5'-AMP and non-absorbing electrolyte (boric acid), as well as the applied voltage, are optimized for resolution, efficiency, and signal-to-noise ratio. A baseline resolution of all nine SMCFAs is achieved in less than 15 min. Additionally, the developed CE-IPD method shows promising potential to identifying SMCFA in rat fecal supernatant. The presented analytical assay is simple, economical, and has considerably good repeatability. The intraday and interday RSD of less than 1 and 2% for relative migration time, as well as less than 14 and 15% for peak area, respectively, were obtained for SMCFA in fecal solution.

Sex-dependent regulation of social reward by oxytocin: an inverted U hypothesis.

The rewarding properties of social interactions are essential for the expression of social behavior and the development of adaptive social relationships. Here, we review sex differences in social reward, and more specifically, how oxytocin (OT) acts in the mesolimbic dopamine system (MDS) to mediate the rewarding properties of social interactions in a sex-dependent manner. Evidence from rodents and humans suggests that same-sex social interactions may be more rewarding in females than in males. We propose that there is an inverted U relationship between OT dose, social reward, and neural activity within structures of the MDS in both males and females, and that this dose-response relationship is initiated at lower doses in females than males. As a result, depending on the dose of OT administered, OT could reduce social reward in females, while enhancing it in males. Sex differences in the neural mechanisms regulating social reward may contribute to sex differences in the incidence of a large number of psychiatric and neurodevelopmental disorders. This review addresses the potential significance of a sex-dependent inverted U dose-response function for OT's effects on social reward and in the development of gender-specific therapies for these disorders.

Sex-dependent regulation of social reward by oxytocin receptors in the ventral tegmental area.

Social reward is critical for social relationships, and yet we know little about the characteristics of social interactions that are rewarding or the neural mechanisms underlying that reward. Here, we investigate the sex-dependent role of oxytocin receptors within the ventral tegmental area (VTA) in mediating the magnitude and valence of social reward. Operant and classical conditioning tests were used to measure social reward associated with same-sex social interactions. The effects of oxytocin, selective oxytocin receptor agonists, antagonists, and vehicle injected into the VTA on social reward was determined in male and female Syrian hamsters. The colocalization of FOS and oxytocin in sites that project to the VTA following social interaction was also determined. Females find same-sex social interactions more rewarding than males and activation of oxytocin receptors in the VTA is critical for social reward in females, as well as males. These studies provide support for the hypothesis that there is an inverted U relationship between the duration of social interaction and social reward, mediated by oxytocin; and that in females the dose-response relationship is initiated at lower doses compared with males. Same-sex social interaction is more rewarding in females than in males, and an inverted U relationship mediated by oxytocin may have a critical role in assigning positive and negative valence to social stimuli. Understanding these sex differences in social reward processing may be essential for understanding the sex differences in the prevalence of many psychiatric disorders and the development of gender-specific treatments of neuropsychiatric disorders.

ARC and BDNF expression after cocaine self-administration or cue-induced reinstatement of cocaine seeking in adolescent and adult male rats.

Recreational drug use peaks during adolescence. Our research with adolescent vs adult male rats, however, shows that rats taking cocaine as adolescents have lower levels of cue-induced reinstatement of drug-seeking than adults, despite similar levels of intravenous (i.v.) cocaine self-administration. Lower rates of reinstatement in younger rats could be explained by higher levels of brain plasticity. Two neuroplasticity-related genes, activity-regulated cytoskeletal-associated gene (Arc) and brain-derived neurotrophic factor (Bdnf), influence cocaine self-administration and cue-induced reinstatement. We tested whether reinstatement of cocaine seeking correlates with expression of these genes in reinforcement-related brain regions. Adolescent and adult male rats (postnatal day 35 or 83-95 at start) were allowed to acquire lever-pressing maintained by i.v. infusions of cocaine in daily 2-h sessions over 13 days. At one of three experimental time points, rats were sacrificed and tissue collected to analyze Arc and Bdnf mRNA by in situ hybridization in the entire medial prefrontal cortex and entire nucleus accumbens, as well as relevant subregions: prelimbic cortex, infralimbic cortex, and nucleus accumbens core and shell. Despite taking similar amounts of cocaine, adolescents reinstated less than adults. Gene expression was most notable in the prelimbic cortex, was generally higher in adolescent-onset groups, and was higher with longer abstinence. These data partially support the hypothesis that higher levels of Arc and/or Bdnf gene expression in reinforcement-related brain regions of younger animals contribute to lower rates of extinction responding and/or reinstatement. Future studies should include mechanistic analysis of Arc, Bdnf, and their signaling pathways in age-dependent effects of cocaine.

Role of oxytocin in the ventral tegmental area in social reinforcement.

The rewarding properties of social interactions play a critical role in the development and maintenance of social relationships, and deficits in social reward are associated with various psychiatric disorders. In the present study, we used a novel Operant Social Preference (OSP) task to investigate the reinforcing properties of social interactions under conditions of high or low reward value, and high or low behavioral effort in male Syrian hamsters. Further, we investigated the role of oxytocin (OT) in a key structure of the mesolimbic reward system, the ventral tegmental area (VTA), in mediating the reinforcing properties of social interaction. Adult male hamsters were placed in a three-chambered apparatus, and allowed access to either a social chamber containing an unrestrained conspecific or a non-social chamber, by pushing through a one-way entry, vertical-swing door. Increasing the duration of social interaction (reward value) decreased the frequency of entering the social interaction chambers, whereas decreasing the duration of social interaction conversely increased the frequency of entries. Moreover, increasing behavioral effort required to access social interaction decreased the frequency of entries, especially under conditions when the duration of social interaction was only 5 s. OT injected into the VTA decreased the frequency of entering social interaction chambers in a manner similar to that observed when duration was increased, whereas injection of an OT receptor antagonist in the VTA increased the frequency of seeking social interaction. Taken together, these data support the hypothesis that activation of OT receptors in the VTA are critical for the reinforcing properties of social interactions. Furthermore, social interactions may exhibit duration and cost dependent reinforcing effects on behavior similar to those observed with food and drugs of abuse.

Abstinence environment contributes to age differences in reinstatement of cocaine seeking between adolescent and adult male rats.

RATIONALE: Extinction responding and cue-induced reinstatement of cocaine seeking after 60-days of forced abstinence are attenuated in male rats that self-administered cocaine during adolescence, compared with adults. Given that environmental enrichment during abstinence decreases reinstatement among adults, a possible explanation for attenuated reinstatement among adolescents is that standard pair-housing in prior studies creates a more stimulating environment for younger rats. OBJECTIVE: Therefore, we tested whether standard pair-housing is necessary for the attenuated reinstatement among adolescents by determining whether an impoverished environment during abstinence would increase reinstatement among adolescents, up to adult levels. Conversely, we also tested whether environmental enrichment could further decrease reinstatement among adolescents, and whether we could replicate effects of environmental enrichment to decrease reinstatement among adults down to adolescent levels (positive controls). METHODS: Adolescent and adult male Wistar rats self-administered cocaine intravenously for 12days (fixed ratio 1; 0.36mg/kg per infusion; 2h sessions). Rats were then moved into enriched (grouped, large cages, novel toys), standard (pair-housed, shoebox cages), or impoverished (isolated, hanging cages) housing conditions. After 60days, extinction and cue-induced reinstatement of cocaine seeking were tested, followed by drug-primed reinstatement (0, 5, 10mg/kg cocaine, i.p.). RESULTS: Consistent with previous results, extinction and cue-induced reinstatement were attenuated in adolescent-onset groups compared with adults; this age difference also extended to drug-primed reinstatement. In support of the present hypothesis, an impoverished environment during abstinence increased reinstatement among adolescents to levels that were not different from adult standard-housing levels. CONCLUSIONS: These data suggest that abstinence environment influences the enduring effects of cocaine among adolescents as well as adults.

A novel operant task to assess social reward and motivation in rodents.

BACKGROUND: Social reward plays a critical role in the development of beneficial social relationships, and disorders of the mechanisms controlling social reward are involved in the etiology of many psychiatric diseases. NEW METHOD: We present a novel operant social preference task to quantify social reward in rodents using an apparatus with three chambers separated by one-way vertical-swing doors. The experimental animal is placed in the larger chamber while the two smaller chambers either remain empty or contain a stimulus animal or other potential reward stimulus. Adding weights to the door can alter effort required for rewards. RESULTS: Hamsters (Mesocricetus auratus) entered the chamber containing a stimulus hamster significantly more frequently than an empty chamber. When the reinforcing effects of social interactions were compared to food reward under progressive cost requirements, the reinforcing effects of social interaction and sunflower seeds were similar. Progressively increasing the door weight decreased number of entries, but increased time spent attempting to open the doors. COMPARISON WITH EXISTING METHODS: The quantification of the rewarding properties of social interactions has almost exclusively used the conditioned place preference (CPP) paradigm. Although robust and reliable, CPP includes a memory component, because it relies on the association of place with the social interaction while the operant task presented here does not. CONCLUSIONS: This task allows for detailed and direct assessment of social and non-social rewards that may serve as effective behavioral reinforcers in this operant conditioning model, and it can be used to investigate the neural mechanisms regulating motivation.

Altered sucrose self-administration following injection of melanocortin receptor agonists and antagonists into the ventral tegmental area.

RATIONALE AND OBJECTIVES: Alpha-melanocyte stimulating hormone (αMSH) and agouti-related protein (AgRP) are antagonistic neuropeptides that play an important role in the control of feeding and body weight through their central actions on the melanocortin-3 and melanocortin-4 receptors. Increasing evidence indicates that αMSH and AgRP can interact with the mesolimbic dopamine system to regulate feeding as well as other behaviors. For example, we have shown previously that injection of melanocortin receptor agonists and antagonists into the ventral tegmental area (VTA) alters both normal home-cage feeding and the intake of sucrose solutions, but it remains unknown whether αMSH and AgRP can also act in the VTA to affect reward-related feeding. METHODS: We tested whether injection of the melanocortin receptor agonist, MTII, or the melanocortin receptor antagonist, SHU9119, directly into the VTA affected operant responding maintained by sucrose pellets in self-administration assays. RESULTS: Injection of MTII into the VTA decreased operant responding maintained by sucrose pellets on both fixed ratio and progressive ratio schedules of reinforcement, whereas SHU9119 increased operant responding under fixed ratio, but not progressive ratio schedules. MTII also increased and SHU9119 decreased 24-h home-cage food intake. CONCLUSIONS: This study demonstrates that αMSH and AgRP act in the VTA to affect sucrose self-administration. Thus, it adds critical information to the growing literature showing that in addition to their well-characterized role in controlling "need-based" feeding, αMSH and AgRP can also act on the mesolimbic dopamine system to control reward-related behavior.

A Comparison of Internal Dispositions and Career Trajectories after Collaborative versus Apprenticed Research Experiences for Undergraduates.

Undergraduate research experiences confer benefits on students bound for science, technology, engineering, and mathematics (STEM) careers, but the low number of research professionals available to serve as mentors often limits access to research. Within the context of our summer research program (BRAIN), we tested the hypothesis that a team-based collaborative learning model (CLM) produces student outcomes at least as positive as a traditional apprenticeship model (AM). Through stratified, random assignment to conditions, CLM students were designated to work together in a teaching laboratory to conduct research according to a defined curriculum led by several instructors, whereas AM students were paired with mentors in active research groups. We used pre-, mid-, and postprogram surveys to measure internal dispositions reported to predict progress toward STEM careers, such as scientific research self-efficacy, science identity, science anxiety, and commitment to a science career. We are also tracking long-term retention in science-related career paths. For both short- and longer-term outcomes, the two program formats produced similar benefits, supporting our hypothesis that the CLM provides positive outcomes while conserving resources, such as faculty mentors. We discuss this method in comparison with course-based undergraduate research and recommend its expansion to institutional settings in which mentor resources are scarce.

Attenuated effects of experimenter-administered heroin in adolescent vs. adult male rats: physical withdrawal and locomotor sensitization.

OBJECTIVES: Early onset of heroin use during adolescence might increase chances of later drug addiction. Prior work from our laboratory suggests, however, that adolescent male rats are actually less sensitive than adults to some enduring effects of heroin self-administration. In the present study, we tested two likely correlates of sensitivity to behavioral reinforcement in rats: physical withdrawal and locomotor sensitization. METHODS: Adolescent (35 days old at start) and adult (79 days old) male Sprague-Dawley rats were administered escalating doses of heroin, increasing from 1.0 to 8.0 mg/kg (i.p.) every 12 h, across 13 days. Somatic signs of spontaneous withdrawal were scored 12 and 24 h after the last injection, and then every 24 h for 5 days; locomotion was recorded concurrently. Challenge injections of heroin (1 mg/kg i.p.) were given at four points: as the first of the escalating doses (day 1), at days 7 and 13 during the escalating regimen, and after 12 days of forced abstinence. Body mass and food intake were measured throughout experimentation. RESULTS: A heroin withdrawal syndrome was not observed among adolescents as it was among adults, including somatic signs as well as reduced locomotion, body mass, and food intake. On the other hand, heroin-induced locomotor sensitization did not differ across ages. CONCLUSION: Reduced withdrawal is consistent with the attenuated reinforcing effects of heroin among adolescent male rats that we reported previously. Thus, it is possible that adolescent rats could reveal important neuroprotective factors for use in treatment of heroin dependence.

A role for the prefrontal cortex in heroin-seeking after forced abstinence by adult male rats but not adolescents.

Adolescent drug abuse is hypothesized to increase the risk of drug addiction. Yet male rats that self-administer heroin as adolescents show attenuated drug-seeking after abstinence, compared with adults. Here we explore a role for neural activity in the medial prefrontal cortex (mPFC) in age-dependent heroin-seeking. Adolescent (35-day-old at start; adolescent-onset) and adult (86-day-old at start) male rats acquired lever-pressing maintained by heroin using a fixed ratio one reinforcement schedule (0.05 and 0.025 mg/kg per infusion). Following 12 days of forced abstinence, rats were tested for heroin-seeking over 1 h by measuring the number of lever presses on the active lever. Unbiased stereology was then used to estimate the number of Fos-ir(+) and Fos-ir(-) neurons in prelimbic and infralimbic mPFC. As before, adolescents and adults self-administered similar amounts of heroin, but subsequent heroin-seeking was attenuated in the younger rats. Similarly, the adolescent-onset group failed to show significant neural activation in the prelimbic or infralimbic mPFC during the heroin-seeking test, whereas the adult-onset heroin self-administration group showed two to six times more Fos-ir(+) neurons than their saline counterparts in both mPFC subregions. Finally, the overall number of neurons in the infralimbic cortex was greater in rats from the adolescent-onset groups than adults. The mPFC may thus have a key role in some age-dependent effects of heroin self-administration.

Heroin self-administration and reinstatement of heroin-seeking in adolescent vs. adult male rats.

RATIONALE: Heroin abuse is prevalent among teenagers, and early onset drug use might predict long-term drug dependence. However, adolescent sensitivity to drug reinforcement has not been explored thoroughly in animal models. OBJECTIVES: This study aimed to compare intravenous (i.v.) self-administration of heroin, as well as extinction and reinstatement of heroin-seeking, in adolescent vs. adult male rats. METHODS: Adolescent (35 days old at start) and adult (86 days old at start) male Sprague-Dawley rats spontaneously acquired lever pressing maintained by i.v. heroin infusions. In experiment 1, self-administration was tested on a fixed ratio 1 schedule of reinforcement (0.05 and 0.025 mg/kg per infusion), followed by within-session extinction and reinstatement tests after 1 or 12 days of abstinence. In experiment 2, self-administration was tested on a progressive ratio schedule (0.0125-0.1 mg/kg per infusion), followed 12 days later by a single test of extinction responding in the presence of cues. RESULTS: In experiment 1, adolescent rats self-administered more heroin than adults. After 1 or 12 days of abstinence, adolescents exhibited less heroin-seeking than adults, although levels of heroin-seeking increased over abstinence period for both age groups. In experiment 2, adolescents and adults reached the same maximal response ratio (breakpoint), although adolescents earned more infusions when response requirements were low. For extinction responding in the presence of cues, heroin-seeking was similar across ages. CONCLUSIONS: Lower levels of heroin-seeking suggest that younger rats are less sensitive than adults to some residual effects of heroin intake.

New perspectives on adolescent motivated behavior: attention and conditioning.

Adolescence is a critical transition period, during which fundamental changes prepare the adolescent for becoming an adult. Heuristic models of the neurobiology of adolescent behavior have emerged, promoting the central role of reward and motivation, coupled with cognitive immaturities. Here, we bring focus to two basic sets of processes, attention and conditioning, which are essential for adaptive behavior. Using the dual-attention model developed by Corbetta and Shulman (2002), which identifies a stimulus-driven and a goal-driven attention network, we propose a balance that favors stimulus-driven attention over goal-driven attention in youth. Regarding conditioning, we hypothesize that stronger associations tend to be made between environmental cues and appetitive stimuli, and weaker associations with aversive stimuli, in youth relative to adults. An attention system geared to prioritize stimulus-driven attention, together with more powerful associative learning with appetitive incentives, contribute to shape patterns of adolescent motivated behavior. This proposed bias in attention and conditioning function could facilitate the impulsive, novelty-seeking and risk-taking behavior that is typical of many adolescents.

A high fructose diet does not affect amphetamine self-administration or spatial water maze learning and memory in female rats.

High energy diets can have a detrimental effect on brain plasticity. For example, a high fructose diet impairs spatial memory in male rats. The aim of the present study was to determine whether a high fructose diet impairs another form of learning and memory: drug reinforcement learning. Female Sprague-Dawley rats were fed a high fructose diet (60%) from weaning at postnatal day (PND) 21, then allowed to acquire lever-pressing maintained by intravenous (i.v.) amphetamine at PND 68, 109, or 165. Acquisition was tested on a fixed ratio one (FR1) schedule of reinforcement (0.025 mg/kg/infusion, 1h daily sessions, 10 sessions over 14 days), followed by testing for reinforcing efficacy on a progressive ratio (PR) schedule (0.025, 0.01, and 0.1mg/kg/infusion), 14 days of abstinence, and within-session extinction and reinstatement tests. Subsequently, water maze acquisition and retention were tested in these subjects as well as a separate cohort tested in the water maze only. The diet had no effect on acquisition, reinforcing efficacy, extinction, or reinstatement of amphetamine seeking. Nor did the diet alter any measures of spatial memory. The high fructose diet did decrease body mass and increase relative liver and spleen mass, but did not affect plasma triglyceride concentrations consistently. Together with prior research on males, these results suggest that the metabolism of fructose and the effects of a high fructose diet on learning and memory may be sex-dependent.

Time-dependent increases in cue-induced reinstatement of sucrose seeking after sucrose self-administration in adolescence.

Previously we reported that time-dependent increases in cue-induced reinstatement of cocaine seeking were attenuated in rats self-administering cocaine as adolescents, compared with adults (Li and Frantz (2009)). Now using sucrose self-administration, we report time-dependent increases in cue-induced reinstatement of sucrose seeking are similar across age groups, suggesting that age differences in reinstatement of cocaine seeking depend on specific effects of cocaine, not a compromised ability among younger rats to associate cues with rewards.

Attenuated incubation of cocaine seeking in male rats trained to self-administer cocaine during periadolescence.

RATIONALE AND OBJECTIVES: Although onset of drug use during adolescence appears to increase long-term vulnerability to drug dependence in humans, relatively little is known about extinction and reinstatement of drug seeking after periadolescent onset of drug self-administration in laboratory animals. Furthermore, although cue-induced reinstatement of cocaine seeking increases progressively during abstinence from cocaine self-administration in adult subjects, this "incubation of cocaine craving" remains unexplored after adolescent drug intake in animal models. MATERIALS AND METHODS: We allowed periadolescent (postnatal day (PND) 35 at start) and adult (PND 83-95 at start) male Wistar rats to self-administer cocaine (0.36 mg/kg/infusion) in 2-h daily sessions on a fixed ratio 1 schedule of reinforcement over 14 days. Then, we compared extinction and cue-induced or cocaine priming-induced reinstatement (10 mg/kg cocaine, intraperitoneal) of cocaine seeking in both age groups after 30 days of abstinence in home cages. In separate cohorts, we tested for time-dependent increases in cue-induced reinstatement over approximately 1, 14, 30, or 60 days of abstinence in both age groups. RESULTS: Adolescent and adult rats self-administered similar amounts of cocaine. Subsequent cue-induced reinstatement was lower in the adolescent-onset group after a 30-day abstinence period, but cocaine priming-induced reinstatement did not differ across ages. Also, extinction responding and time-dependent increases in cue-induced reinstatement (incubation) were less pronounced in rats that took cocaine as adolescents compared with adults. CONCLUSIONS: Surprisingly, these results may reflect resistance among adolescent subjects to some enduring effects of drug self-administration, such as reward learning.