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Expression data from the basolateral amygdala of Long-Evans rats with a history of limited intermittent sucrose snacks

ABSTRACT: To study the molecular mediators of naturally rewarding effects of palatable food we used a model of palatable “snacking” (Ulrich-Lai et al., 2007) in which rats are given chronic, brief access to a limited amount of sucrose solution (30%). Single housed, male Long-Evans rats (250g) (n=12 per group) from Harlan Labs (Indianapolis, IN) received normal rat chow (Harlan Teklad) and water ad libitum for the duration of the experiment. After a one-week period of acclimation, rats were randomly assigned to drink treatment groups of either 30% sucrose solution or water. Rats received a 14-day regimen of twice daily (9:30 and 15:30) brief (maximum of 30 minutes) limited (up to 4 mL) access of their assigned drink solution. Drink solutions were delivered via a graduated sipper placed onto the cage top in addition to the existing water bottle and sippers were immediately removed when the animal had consumed 4mL or after the 30-minute access period, whichever occurred first. Drink intake, food intake, and body weight were monitored throughout the experiment to verify that the rats learned to drink sucrose, that they adjusted chow intake for calories consumed from sucrose (~10%), and that there was no effect on body weight gain as is normally seen with this model (Ulrich-Lai et al., 2007). Drink treatment terminated on day 14 and at 8:00 on the morning of day 15, the rats were sacrificed by rapid decapitation. BLA tissue was dissected, RNA extracted, and gene expression changes between water and sucrose groups were accessed by microarray.

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE20344 | GEO | 2010/02/17

SECONDARY ACCESSION(S): PRJNA125423

REPOSITORIES: GEO

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Project description:To study the molecular mediators of naturally rewarding effects of palatable food we used a model of palatable âsnackingâ (Ulrich-Lai et al., 2007) in which rats are given chronic, brief access to a limited amount of sucrose solution (30%). Single housed, male Long-Evans rats (250g) (n=12 per group) from Harlan Labs (Indianapolis, IN) received normal rat chow (Harlan Teklad) and water ad libitum for the duration of the experiment. After a one-week period of acclimation, rats were randomly assigned to drink treatment groups of either 30% sucrose solution or water. Rats received a 14-day regimen of twice daily (9:30 and 15:30) brief (maximum of 30 minutes) limited (up to 4 mL) access of their assigned drink solution. Drink solutions were delivered via a graduated sipper placed onto the cage top in addition to the existing water bottle and sippers were immediately removed when the animal had consumed 4mL or after the 30-minute access period, whichever occurred first. Drink intake, food intake, and body weight were monitored throughout the experiment to verify that the rats learned to drink sucrose, that they adjusted chow intake for calories consumed from sucrose (~10%), and that there was no effect on body weight gain as is normally seen with this model (Ulrich-Lai et al., 2007). Drink treatment terminated on day 14 and at 8:00 on the morning of day 15, the rats were sacrificed by rapid decapitation. BLA tissue was dissected, RNA extracted, and gene expression changes between water and sucrose groups were accessed by microarray. We used microarray to assess the changes in gene expression in the BLA of rats that have had a history of sucrose snacks (n=12) vs control (n=12). Rats received access (up to 4 mL) to extra bottles containing water (control) or 30% sucrose twice daily for 14 days. Rats had ab lib access to chow and water for the duration of the experiment. Rats were sacrificed the morning of day 15 and the BLA was removed for microarray analysis. Functional cluster analysis was performed on the list of 145 significantly upregulated genes using Ingenuity Pathway Analysis (Ingenuity Systems, Redwood City, CA). See supplementary file at the foot of this record.

Project description:Analysis of gene expression profiles of epididymal fat from DIO rats; We applied a comparative functional genomics approach to evaluate diet-induced obese (DIO) rats as an obesity model Experiment Overall Design: Gene expression profiles from DIO and lean rats were generated and compared. A private colony of male Long-Evans rats was set up at Harlan (Harlan-Sprague Dawley, Indianapolis, IN). The animals had been on either a high fat/high sucrose diet (TD95217, Harlan Takled, Madison, WI) or a regular chow (Purina 5008; Ralston-Purina, St. Louis, MO) since weaning. Rats were shipped at 14-weeks of age to our facility and maintained on a 12:12-hour light-dark photoperiod (light on 10:00 PM). They were individually housed throughout the study at ambient temperature, and had free access to diets and water. After 2 weeks acclimation to the facility, body weight of the rats and weight of food consumed in the last 7 days were measured once a week. Body fat mass was analyzed by nuclear magnetic resonance (NMR) using an Echo Medical System (Houston, TX) instrument 3 times throughout the study. Fat free mass is the difference between body mass and fat mass. All animal studies were conducted in compliance with approved institutional animal care and use protocols according to NIH guidelines (NIH publication No. 86-23, 1985). OGTT was conducted after an overnight fasting when animals were 21 weeks of age. After a baseline blood collection (time 0), animals were dosed orally with 1.5 gram/kg glucose. Subsequence blood samples were collected at 30, 60, and 120 minutes post glucose challenge for the determination of plasma glucose and insulin levels. At 25 weeks of age, blood was collected by tail bleeding after overnight fasting. Rats were killed a week later at 09:00am by rapid decapitation. Trunk blood was collected and epididymal fat tissue excised. Various circulating metabolic parameters were measured under fasted (week 25) and fed conditions (week 26). Total RNA from rat epididymal fat was isolated with RNA STAT-60 (Tel-Test) according to the manufacturer's protocol. 5 micrograms of total RNA were labeled and hybridized to Affymetrix RAE230_2 arrays according to the Affymetrix protocol.

Project description:By using high-density oligonucleotide arrays, we profiled gene expression in reward-related brain regions of rats that developed escalated cocaine intake after extended access to cocaine (6 h per day). Rats allowed restricted daily access to cocaine (only 1 h) that displayed a stable level of cocaine intake and cocaine naive rats were used for controls. Four analysis methods were compared: Affymetrix microarray suite 4 and microarray suite 5, which use perfect-match-minus-mismatch models, and dchip and rma, which use perfect-match-only models to generate expression values. Results were validated by RT-PCR in individual animals from an independent replication of the experiment. A small number of genes was associated with escalated cocaine intake (ESC genes). Unexpectedly, of the brain regions examined [prefrontal cortex, nucleus accumbens, septum, lateral hypothalamus (LH), amygdala, and ventral tegmental area], the LH was the most transcriptionally responsive in escalation of cocaine intake. Most of the ESC genes identified are also expressed during synaptogenesis and synaptic plasticity and include genes that code for several presynaptic and postsynaptic proteins involved in neurotransmission. These results suggest that LH intrinsic circuitry undergoes a structural reorganization during escalation of cocaine use. This remodeling of LH circuitry could contribute to the chronic deficit in reward function that has been hypothesized to drive the transition to drug addiction. Results also support the value of using multiple analysis strategies to identify the most robust changes in gene expression and to compensate for the biases that affect each strategy. Experiment Overall Design: The experimental design for this study comprised three groups of rats (n = 8) with different daily access to a continuous schedule of i.v. cocaine self-administration (0.25 mg per injection): 0 h access per day (drug-naive control rats), 1 h access per day (ShA rats), or 6 h access per day (LgA rats). Experiment Overall Design: Gene expression profiling was then performed for each dissected brain region by using Affymetrix rat neurobiology arrays.

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Expression data from the basolateral amygdala of Long-Evans rats with a history of limited intermittent sucrose snacks

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