Jeong Won Jahng

Chronic food restriction in young rats results in depression- and anxiety-like behaviors with decreased expression of serotonin reuptake transporter

Evidence of semi-starvation is commonly found in patients with eating disorders. This study was conducted to examine the adverse effects of chronic caloric restriction in young rats, since there have been increasing incidence of eating disorders especially among young populations. Food restriction group was supplied daily with 50% of chow consumed by its ad libitum fed control group from postnatal day 28. After 5 weeks of food restriction, brain contents of serotonin (5-hydroxy-tryptamine; 5-HT) and its metabolite 5-hydroxyindol acetic acid were analyzed by high-performance liquid chromatography and mRNA expression of 5-HT reuptake transporter (5-HTT) by in situ hybridization. Plasma corticosterone levels were determined by radioimmunoassay. Behavioral assessments were performed with Porsolt swim test for depressive behavior and with elevated plus maze test for anxiety. Five weeks of food restriction markedly increased plasma level of corticosterone, and significantly decreased 5-HT turnover rates in the hippocampus and the hypothalamus. 5-HTT mRNA expression decreased in the raphe nucleus of food restricted rats compared with free fed controls. Immobility time during the swim test increased in the food restricted group, compared to the control group. Food restricted rats spent more time in the closed arms, less time in the open arms, of elevated plus maze compared with control rats. These results suggest that chronic caloric restriction in young rats may lead to the development of depressive and/or anxiety disorders, likely, in relation with dysfunction of brain 5-HT system.

Dexamethasone reduces food intake, weight gain and the hypothalamic 5-HT concentration and increases plasma leptin in rats.

This study was conducted to define the regulatory mechanisms underlying stress-induced decreases in food intake and weight gain. Rats received a single or 4 daily injections of dexamethasone (0.1 or 1 mg/kg). Food intake and weight gain were recorded, and plasma leptin, brain contents of serotonin (5-hydroxytryptamine; 5-HT), 5-hydroxy-indole-acetic acid (5-HIAA) and the raphe expression of tryptophan hydroxylase (TPH), monoamine oxidase A (MAO-A) and 5-HT reuptake transporter (5-HTT) genes were examined. A single injection of dexamethasone did not acutely affect food intake, but cumulative food intake and weight gain were suppressed dose-dependently by daily injections of dexamethasone. Both a single and repeated injections of dexamethasone elevated plasma leptin in a dose dependent manner. 5-HT contents in the hypothalamus was decreased, but 5-HIAA increased, both by a single and repeated dexamethasone. A single injection of dexamethasone did not affect mRNA expressions of TPH, MAO-A and 5-HTT genes, but repeated dexamethasone increased them in the dorsal raphe nucleus. These results suggest that plasma leptin may play a role in dexamethasone-induced anorexia. Additionally, increased expression of MAO-A and 5-HTT genes by repeated dexamethasone appears to be implicated in decreases of the brain 5-HT contents.

Post-weaning isolation promotes food intake and body weight gain in rats that experienced neonatal maternal separation.

Neonatal maternal separation (MS) in rats has been reported to result in permanent dysfunctions of the hypothalamic-pituitary-adrenal axis and the development of anxiety- and depression-like behaviors later in life. In this study, we examined the effects of post-weaning social isolation stress on food intake and body weight gain of rats with MS experience. MS was performed daily for 180 min during the first 2 weeks of birth and nonhandled control (NH) pups were left undisturbed. Weanling male pups were caged either in a group of three or singly (social isolation), and then subjected to behavioral sessions for anxiety- or depression-like behaviors at 2 months of age. Social isolation following MS experience, but neither MS nor social isolation alone, significantly increased food intake and weight gain. MS pups showed increased immobility in forced swim test, compared to NH pups, regardless of their housing conditions. In elevated plus maze test, group-caged MS pups spent less time in the open arms and more time in the closed arms than group-caged NH pups, but social isolation did not further affect the arm stay of MS pups. However, statistical analyses revealed an interaction between MS and social isolation not only in the time spent in each arms, but also in defecation scores during the ambulatory activity test. These results suggest that post-weaning social isolation may promote hyperphagia and weight gain in young rats that experienced neonatal maternal separation, perhaps, in relation with its impact on the psycho-emotional behaviors of MS pups.

An animal model of eating disorders associated with stressful experience in early life

Experience of childhood abuse is prevalent among patients with eating disorders, and dysfunction of the hypothalamic-pituitary-adrenal (HPA) axis is implicated in its pathophysiology. Neonatal maternal separation is considered as an animal model of stressful experience early in life. Many of studies have demonstrated its impact both on the activity of HPA axis and the development of psycho-emotional disorders later in life. In this paper, a series of our researches on developing an animal model of eating disorders is reviewed. An animal model of neonatal maternal separation was used; Sprague-Dawley pups were separated from dam daily for 180 min during the first 2 weeks of life (MS) or undisturbed. Anxiety-/depression-like behaviors were observed in MS rats at the age of two months with decreased serotonergic activity in the hippocampus and the raphe. Post-weaning social isolation promoted food intake and weight gain of adolescent MS pups, with impacts on anxiety-like behaviors. Sustained hyperphagia was observed in the MS pups subjected to a fasting/refeeding cycle repeatedly during adolescence, with increased plasma corticosterone levels. Anhedonia, major symptom of depression, to palatable food was observed in adolescent MS pups with blunted response of the mesolimbic dopaminergic activity to stress. Results suggest that neonatal maternal separation lead to the development of eating disorders when it is challenged with social or metabolic stressors later in life, in which dysfunctions in the HPA axis and the brain monoaminergic systems may play important roles.

Fasting-induced increases of arcuate NPY mRNA and plasma corticosterone are blunted in the rat experienced neonatal maternal separation

This study was conducted to examine the effects of neonatal maternal separation on the hypothalamic expression of feeding peptides in later life. Pups in maternal separation (MS) groups were separated from their dam for 3 h daily from postnatal day (PND) 1-14, while pups in non-handled (NH) groups were left undisturbed. Rats were sacrificed on PND 60 to examine the gene expression of neuropeptide Y (NPY) and pro-opiomelanocortin (POMC) in the hypothalamic arcuate nucleus by mRNA in situ hybridization. Half of the rats from each group were food-deprived for 48 h before sacrifice. POMC mRNA expression increased in the free fed MS group compared with the free fed NH group. Food deprivation significantly decreased the arcuate POMC mRNA level in both groups. Body weight gain, basal levels of plasma corticosterone, leptin, and arcuate NPY mRNA were not modulated by experience of neonatal maternal separation. However, fasting-induced increases of plasma corticosterone and arcuate NPY expression were blunted in MS rats. These results suggest that neonatal maternal separation may increase the basal expression level of arcuate POMC mRNA, while inhibit the fasting-induced expression of arcuate NPY mRNA, later in life. Lastly, the altered expression of arcuate NPY mRNA, but not of arcuate POMC mRNA, appeared to be related with altered activity of the hypothalamic-pituitary-adrenal gland axis in offspring by neonatal maternal separation.

Systemic 5-hydroxy-L-tryptophan down-regulates the arcuate CART mRNA level in rats

This study was conducted to determine if serotonin (5-hydroxytryptamine; 5-HT) system correlates with the hypothalamic expression of cocaine-amphetamine-regulated transcript (CART) gene. Rats received intraperitoneal 5-hydroxy-L-tryptophan (5-HTP; a single or three daily injections at a dose of 100 mg/kg/10 ml), and CART mRNA level in the hypothalamus was examined by in situ hybridization at different time points. The 5-HT contents of the hypothalamus as well as the brainstem was increased persistently by 5-HTP injections, and food intake and body weight gain reduced. CART mRNA level decreased significantly in the hypothalamic arcuate nucleus by three daily 5-HTP, but not by a single injection. The pair-fed group of the chronic 5-HTP did not show a decrease in the arcuate CART mRNA level. The plasma leptin level markedly decreased in the chronic 5-HTP group, compared to the saline group, however, still higher than the pair-fed group with a statistical significance. These results suggest that 5-HT may suppress CART mRNA expression in the arcuate nucleus, not only by leptin signaling via its anorectic effect on the control of food intake, but also by some non-leptin mediated pathway.

Refeeding-induced expression of neuronal nitric oxide synthase in the rat paraventricular nucleus

We have previously reported that food deprivation decreases the expression of neuronal nitric oxide synthase (nNOS) in the hypothalamic paraventricular nucleus (PVN) of rats, and this reduction is inhibited by blockade of glucocorticoid receptors. In this study, we examined whether the fasting-induced decrease in nNOS gene expression in the PVN is restored by refeeding. The number of nNOS immunopositive cells in the PVN, which was markedly decreased by 48 h of food deprivation, increased significantly after 6 h of refeeding and was fully restored by 24 h after refeeding. The plasma corticosterone level, which was markedly increased by food deprivation, decreased significantly within 30 min after refeeding and returned to the free fed control level by 6 h. Synthetic glucocorticoid dexamethasone blocked the refeeding-induced nNOS expression in the PVN without suppressing food intake. Refeeding with a non-caloric food mash for 5 h failed to restore the fasting-induced decrease in the PVN-nNOS but did, however, successfully restore the plasma corticosterone level. These results suggest that the refeeding-induced nNOS expression in the PVN is a nutrient-directed event and that plasma glucocorticoids may play an inhibitory role in the regulatory pathway. Additionally, glucocorticoid disinhibition alone does not appear to be sufficient to induce nNOS expression in the PVN; nNOS expression in the PVN upon refeeding may require both nutrient supplementation and glucocorticoid disinhibition.

MK801 increases feeding and decreases drinking in nondeprived, freely feeding rats.

The noncompetitive NMDA receptor antagonist MK801 has been reported to increase food intake in rats during scheduled test meals of palatable foods or after food deprivation, but not in nondeprived rats given rodent chow. To determine if MK801 has an effect on spontaneous meals, MK801 (100 microg/kg) was administered 15 min prior to dark onset to nondeprived rats maintained on powdered rodent chow, and spontaneous food and water access was measured. MK801 increased the length of the first meal and the amount of time spent feeding within the meal. Conversely, MK801 decreased the length and size of the first drinking bout. MK801 did not alter the latency to the first meal or drinking bout, nor the intervals between successive meals or bouts. The effects of MK801 on feeding and drinking bouts were partially confirmed by measuring total chow and water intake over the first 2 h of the dark period. Thus, acute MK801 can significantly alter spontaneous chow feeding and drinking in nondeprived rats when administered prior to dark onset.

Adolescence fluoxetine increases serotonergic activity in the raphe-hippocampus axis and improves depression-like behaviors in female rats that experienced neonatal maternal separation.

This study was conducted to examine if fluoxetine, a selective 5-hydroxytryptamine (5-HT) reuptake inhibitor, would reverse adverse behavioral effects of neonatal maternal separation in female rats. Sprague-Dawley pups were separated from dam daily for 3h during postnatal day (PND) 1-14 (maternal separation; MS) or left undisturbed (non-handled; NH). Female NH and MS pups received intraperitoneal injection of fluoxetine (10mg/kg) or vehicle daily from PND 35 until the end of the whole experimental period. Rats were either subjected to behavioral tests during PND 44-54, or sacrificed for neurochemical analyses during PND 43-45. Daily food intake and weight gain of both NH and MS pups were suppressed by fluoxetine, with greater effects in MS pups. MS experience increased immobility and decrease swimming in forced swim test. Swimming was increased, although immobility was not significantly decreased, in MS females by adolescence fluoxetine. However, adolescence fluoxetine increased immobility during forced swim test and decreased time spent in open arms during elevated plus maze test in NH females. Fluoxetine normalized MS-induced decrease of the raphe 5-HT levels and increased 5-HT metabolism in the hippocampus in MS females, and increased the hypothalamic 5-HT both in NH and MS. Fluoxetine decreased the raphe 5-HT and increased the plasma corticosterone in NH females. Results suggest that decreased 5-HTergic activity in the raphe nucleus is implicated in the pathophysiology of depression-like behaviors, and increased 5-HTergic activities in the raphe-hippocampus axis may be a part of anti-depressant efficacy of fluoxetine, in MS females. Also, an extra-hypothalamic 5-HTergic activity may contribute to the increased anorectic efficacy of fluoxetine in MS females. Additionally, decreased 5-HT in the raphe and elevated plasma corticosterone may be related with fluoxetine-induced depression- and/or anxiety-like behaviors in NH females.

Histamine H1 Receptor Induces Cytosolic Calcium Increase and Aquaporin Translocation in Human Salivary Gland Cells

One of the common side effects of antihistamine medicines is xerostomia (dry mouth). The current consensus is that antihistamine-induced xerostomia comes from an antimuscarinic effect. Although the effect of antihistamines on salivary secretion is both obvious and significant, the cellular mechanism whereby this happens is still unclear because of the lack of knowledge of histamine signaling in human salivary glands. Here, we have studied histamine receptors and the effect of antihistamines on human submandibular acinar cells. In primary cultured human submandibular gland and a HSG cell line, histamine increased the intracellular Ca2+ concentration. The histamine-induced cytosolic free Ca2+ concentration ([Ca2+]i) increase was inhibited by histamine H1 receptor-specific antagonists, and the expression of the functional histamine H1 receptor was confirmed by reverse transcription–polymerase chain reaction. Interestingly, histamine pretreatment did not inhibit a subsequent carbachol-induced [Ca2+]i rise without “heterologous desensitization.” Chlorpheniramine inhibited a carbachol-induced [Ca2+]i increase at a 100-fold greater concentration than histamine receptor antagonism, whereas astemizole and cetrizine showed more than 1000-fold difference, which in part explains the xerostomia-inducing potency among the antihistamines. Notably, histamine resulted in translocation of aquaporin-5 to the plasma membrane in human submandibular gland cells and green fluorescent protein-tagged aquaporin-5 expressing HSG cells. We found that histidine decarboxylase and the histamine H1 receptor are broadly distributed in submandibular gland cells, whereas choline acetyltransferase is localized only at the parasympathetic terminals. Our results suggest that human salivary gland cells express histamine H1 receptors and histamine-synthesizing enzymes, revealing the cellular mechanism of antihistamine-induced xerostomia.