Preference for corn oil in olfactory-blocked mice in the conditioned place preference test and the two-bottle choice test.

We studied the effects of olfactory stimuli on preference for corn oil in mice. In the conditioned place preference test, voluntary intake of 100% corn oil by both olfactory normal and ZnSO4-induced olfactory-blocked (anosmic) mice resulted in their place preference for the corn oil-related box. In the olfactory normal mice, place preference was also observed by voluntary intake of linoleic acid as well as of corn oil. In the two-bottle choice test, normal mice showed significant preference for test fluids that contained corn oil at all concentrations (1-10%) tested relative to vehicle alone. However, the lower concentrations (1 and 3%) of corn oil were not preferred in the anosmic mice. These results suggested that stimuli other than olfaction contributed to the rewarding effects of corn oil, but at lower concentrations olfactory stimuli might act as a signal for the oil.

Opioidergic contribution to conditioned place preference induced by corn oil in mice.

We previously reported that voluntary intake of corn oil in the light box showed place preference in the conditioned place preference (CPP) test in mice. In the present study, we investigated the contribution of opioidergic systems to the corn oil-induced CPP in mice. Acquisition of the place preference by corn oil intake was blocked by i.p. injections of an opioid mu antagonist, naloxone (0.1 and 0.3 mg/kg), and delta antagonists, 7-benzylidenenaltrexone (0.5 mg/kg) and naltriben (0.5 mg/kg) 15 min before conditioning. The opioid kappa agonist U-50488H (1 and 3 mg/kg i.p.) also blocked corn oil-induced CPP. Naloxone (1 mg/kg, i.p.) and naltriben (0.5 mg/kg, i.p.) did not affect corn oil intake in the home cage. However, 7-benzylidenenaltrexone (0.5 mg/kg, i.p.) and U-50488H (1 mg/kg i.p.) decreased and increased the corn oil intake, respectively. These results suggested that the rewarding effects of corn oil in the CPP test are at least partially mediated via opioidergic systems through mu and delta receptors. Further, we showed that an opioid kappa agonist reduced the rewarding effects of corn oil in the CPP test in mice, although it increased corn oil intake.

Long-term optional ingestion of corn oil induces excessive caloric intake and obesity in mice.

Corn oil is well tolerated by mice but tolerance may decrease with excessive ingestion. In the present study, we compared the effects of optional ingestion of excessive corn oil with ingestion of water (control) or a 20% sucrose solution in mice. During the entire study, mice consistently ingested 100% corn oil and incrementally ingested 20% sucrose. Food intake in the corn-oil group was approximately constant but that in the sucrose group was slightly decreased. Body-weight gains in the corn-oil group were higher than those in the control and sucrose groups. At the end of the study, hepatic hypertrophy and fatty liver were present, especially in the corn-oil group, and the visceral fat of mice fed corn oil increased significantly compared with the other two groups. These results suggest that mice, when given a choice, will continue to overeat corn oil over the long term, inducing excessive caloric intake and obesity.

Biochemical studies of dopaminergic activation by stimuli of corn oil in the oral cavity in mice.

We investigated the effects of corn oil stimuli in the oral cavity on monoaminergic neurones by measuring the amount of monoamines and their metabolites in brains of mice. Intraoral injection of corn oil (0.1 ml) increased dopamine (DA) turnover rate in the cortex 3, 5 and 10 min after injection in mice. A significant increase of DA turnover rate was also observed in midbrain 10 min after the corn oil injection. Turnover rates of noradrenaline (NA) and serotonin (5-HT) were not affected by the corn oil injection in any of the regions or times tested. In the chronic study, DA and 5-HT contents in the left and right cortexes and 5-HT contents in diencephalon in the chronic corn oil-fed group were significantly higher than those in the-control but NA contents were not significantly different between the two groups in all regions. In conclusion, the present results suggested that the stimuli of corn oil in the oral cavity might activate dopaminergic systems in the brain of mice and its chronic stimuli might increase DA contents in the brain. Although 5-HT contents increased in the brains of mice by the chronic corn oil intake, its physiological implication remains to be elucidated.

Grooming behavior in mice induced by stimuli of corn oil in oral cavity.

Mice show a strong preference for corn oil, which was thought to be elicited by stimulation in the oral cavity. Grooming behavior is known to be induced by dopamine D(1) stimulation in rodents. Therefore, we evaluated stimulation by corn oil in the oral cavity and the contribution of D(1) receptors to corn-oil-induced grooming in mice. Intraoral injection (0.1 ml) of corn oil induced grooming behavior similarly to SKF 38393 (10 mg/kg i.p.), a D(1) agonist, and both were antagonized by pretreatment with SCH 23390, a D(1) antagonist. However, a higher dose was needed for antagonism of the corn-oil-induced grooming compared with that induced by SKF 38393. Long-chain fatty acids, their methyl esters and alcohol, their triglycerides, mineral oil and silicone oil but not glycerin, a short-chain triglyceride, xanthan gum solution, or sucrose solution also induced grooming in mice. Xanthan gum solution, which was suggested to mask oil-like texture, attenuated the silicone-oil- but not corn-oil-induced grooming when injected intraorally as a mixture with an equal volume of the oil (50% suspension). The silicone-oil-induced grooming was reduced by SCH 23390 similarly to that induced by corn oil. These results suggested that stimulation by the oil-like texture in the oral cavity in mice induced grooming behavior and that it might be mediated at least partially via D(1) receptors. Moreover, stimuli other than texture might also contribute to the corn-oil-induced grooming.

Effect of a bovine hypothalamic factor on the release and biosynthesis of growth hormone.

Partial purification of growth hormone (GH)-releasing factor (GRF) by acid extraction followed by gel filtration on Sephadex G-25 has been attained from bovine hypothalami. When rat pituitaries were incubated in 2 ml Krebs Ringer-bicarbonate-glucose (KRBG) medium, a stimulatory effect of the GRF fraction on immunoreactive GH (IR-GH) release was observed, while that of the factor neither on GH synthesis nor release of the synthesized GH was demonstrated. Stimulation of the GH release was exerted maximally within 30 min of incubation. Cycloheximide and actinomycin D, at a concentration which inhibited protein and RNA synthesis to less than 5 and 20% of the control, respectively, were without effect on the stimulatory action of the factor on GH release. On the other hand, stimulation of GH synthesis was observed under incubation in 0.3 ml medium with the factor and enhancing effect of the factor on the IR-GH release was undetectable. These results suggest that stimulation of the release and synthesis of GH mediated by the hypothalamic GRF fraction is under influence of the pool size of incubation media.

Extraction and partial purification of prolactin-release stimulating factor in bovine hypothalami.

Partial purification of prolactin-release stimulating factor (PRF) was performed by Sephadex G-25 gel filtration of bovine hypothalamic extracts. PRF activity was evaluated on the basis of the measurement of immunoreactive prolactin released from the isolated rat hemipituitary in vitro. PRF activity was found in the fractions with Kav=0-0.49 and prolactin-release inhibiting activity was also detected in the fractions with Kav=0.69-0.89. The dose-response relationship was established between the partially purified PRF and its activity. The elution position of the partially purified PRF preceded that of TRH on Sephadex G-25. TRH at the dose of 100 nM stimulated the release of TSH in vitro, but not the release of prolactin. These results may indicate that there exists PRF with a relatively high molecular weight in the bovine hypothalamus.