Effects of Fluoxetine Administration on Regional Galanin Expression in Obese Zucker Rat Hypothalamus

Abstract

The aim of the present work was to study the potential involvement of hypothalamic galanin system in the anorectic mechanism of fluoxetine in obese Zucker rats. Male obese Zucker ( fa/fa ) rats were administered fluoxetine (10 mg/kg; i.p.) daily for two weeks. The control group was given 0.9% NaCl solution. Significant decreases in food intake, final body weight and total body fat were observed after fluoxetine treatment. Although fluoxetine-treated rats showed a decrease in urine elimination, this effect was not enough to compensate decreased water intake, leading to dehydration, as showed by decreased body water content. Chronic fluoxetine administration increased the numbers of galanin positively immunostained neural cells in medial and lateral preoptic areas, lateral hypothalamic area and paraventricular nucleus (rostral and magnocellular regions), without changes in dorsomedial, ventromedial, supraoptic, suprachiasmatic and arcuate nuclei. Taken into account that galanin stimulates appetite, these results could represent rather a compensatory response against reduced food intake than a direct anorectic mechanism. Changes in the magnocellular region of the hypothalamic paraventricular nucleus suggest a role for galanin neural circuits at this level in fluoxetine-induced hydro-osmotic impairment.     

Galanin-like peptide (GALP) is a hypothalamic regulator of energy homeostasis and reproduction

Galanin-like peptide (GALP) was discovered in 1999 in the porcine hypothalamus and was found to be a 60 amino acid neuropeptide. GALP shares sequence homology to galanin (1-13) in position 9-21 and can bind to, as well as activate, the three galanin receptor subtypes (GalR1-3). GALP-expressing cells are limited, and are mainly found in the arcuate nucleus of the hypothalamus (ARC) and the posterior pituitary. GALP-positive neurons in the ARC project to several brain regions where they appear to make contact with multiple neuromodulators. These neuromodulators are involved in the regulation of energy homeostasis and reproduction, anatomical evidence that suggests a role for GALP in these physiological functions. In support of this idea, GALP gene expression is regulated by several factors that reflect metabolic state including the metabolic hormones leptin and insulin, thyroid hormones, and blood glucose. Considerable evidence now exists to support the hypothesis that GALP has a role in the regulation of energy homeostasis and reproduction; and, that GALP’s role may be independent of the known galanin receptors. In this review, we (1) provide an overview of the distribution of GALP, and discuss the potential relationship between GALP and other neuromodulators of energy homeostasis and reproduction, (2) discuss the metabolic factors that regulate GALP expression, (3) review the evidence for the role of GALP in energy homeostasis and reproduction, (4) discuss the potential downstream mediators and mechanisms underlying GALP’s effects, and (5) discuss the possibility that GALP may mediate its effects via an as yet unidentified GALP-specific receptor.     

Cardiac ischemia-reperfusion regulates sympathetic neuropeptide expression through gp130-dependent and independent mechanisms

Cardiac function is regulated by a balance of sympathetic and parasympathetic transmission. Neuropeptide Y (NPY) and galanin (GAL) released from cardiac sympathetic neurons inhibits parasympathetic transmission in the heart. Sympathetic peptides may contribute to autonomic imbalance, which is characterized by increased sympathetic and decreased parasympathetic transmission and contributes to life threatening cardiovascular pathologies. Several gp130 cytokines are increased in the heart after myocardial infarction (MI), and these cytokines stimulate neuropeptide expression in sympathetic neurons. We used mice whose sympathetic neurons lack the gp130 receptor (gp130^DBH-Cre/lox mice) to ask if cytokine activation of gp130 regulated neuropeptide expression in cardiac sympathetic nerves after MI. Myocardial infarction decreased NPY mRNA through a gp130 independent mechanism and increased VIP and PACAP mRNA via gp130, while GAL mRNA was unchanged. Immunohistochemistry revealed a gp130-dependent increase in PACAP38 in cells of the stellate ganglion after MI, and PACAP was detected in pre-ganglionic fibers of all genotypes and surgical groups. VIP was identified in a few sympathetic nerve fibers in all genotypes and surgical groups. GAL and PACAP38 were not detected in sham hearts, but peptide immunoreactivity was high in the infarct three days after MI. Surprisingly, peptides were abundant in cells that co-labeled with macrophage markers F4/80 and MAC2, but were not detected in sympathetic axons. PACAP protects cardiac myocytes from apoptosis, and GAL stimulates axon regeneration in addition to inhibiting parasympathetic transmission. Thus, these peptides may play an important role in cardiac and neuronal remodeling after ischemia-reperfusion.     

Model for predicting and phenotyping at normal weight the long-term propensity for obesity in Sprague-Dawley rats

Tests were conducted to determine whether weight gain or nutrient intake measures during the first week of exposure to a macronutrient diet can accurately predict an animal's long-term propensity towards obesity. In multiple groups of normal-weight Sprague-Dawley rats (n=35-70/group), daily weight gain during the first 5 days on a high-fat diet (45-60% fat) was found to be strongly, positively correlated (r=+0.71 to r=+0.82) with accumulated body fat in 4 dissected depots after 4-6 weeks on the diet. This measure consistently identified obesity-prone (OP) rats which, relative to the obesity-resistant (OR) rats, were only slightly heavier (+15 g, 4%) and hyperphagic (+9 kcal, 8%) after 5 days but markedly heavier (+70g) with up to 2-fold greater fat mass after several weeks on the diet. Other dietary conditions and measures revealed weaker relationships to ultimate body fat accrual. The OP rats identified by their 5-day weight-gain score exhibited at this early stage clear disturbances characteristic of markedly obese rats. These included elevated leptin, insulin, triglycerides and glucose, along with increased lipoprotein lipase activity (LPL) in adipose tissue and galanin expression in the paraventricular nucleus. Most notable were significant reductions in muscle of LPL activity and ratio of beta-hydroxyacyl-CoA dehydrogenase to citrate synthase activity, indicating a decline in lipid transport and capacity of muscle to metabolize lipids. By occurring early with initial weight gain, these hypothalamic and metabolic disturbances in OP rats, favoring fat storage in adipose tissue over fat oxidation in muscle, may have causal relationships to long-term accumulation of body fat.     

前列腺组织Galanin及GalR2表达及意义

目的探讨Galanin及Galanin 2型受体（GalR2）在前列腺良恶性组织中的表达及其临床意义。方法收集50例前列腺增生（BPH）、50例未转移前列腺癌（PCa）及30例发生骨转移前列腺癌（BM）标本,应用免疫组织化学法检测Galanin及GalR2的表达,并进行临床病理相关性分析。结果 BPH组、PCa组和BM组Galanin阳性表达分别为9、34、25例;GalR2阳性表达分别为40、7、2例。PCa组及BM组Galanin的阳性表达率显著高于BPH组（χ2=25.50、32.75,P〈0.01）,而PCa组与BM组比较差异无统计学意义（P〉0.05）;BPH组GalR2的阳性表达率显著高于PCa组及BM组（χ2=43.72、40.43,P〈0.01）,而PCa组与BM组比较差异无统计学意义（P〉0.05）。在PCa组及BM组内,Galanin与GalR2的表达均呈显著负相关（rs=-0.55、-0.77,P〈0.01）。结论 Galanin在前列腺癌组织有较高的阳性表达;GalR2在BPH组织有较高的阳性表达,而在恶性肿瘤组织中低表达。Galanin在前列腺癌组织可能通过与GalR2之外的两种受体相结合从而发挥生物学作用;Galanin与GalR2蛋白的联合检测对前列腺良恶性病变的诊断具有较好的临床意义。     

Differential effects of enterostatin, galanin and opioids on high-fat diet consumption

Enterostatin, the activation peptide of pancreatic procolipase, suppresses high-fat diet consumption both centrally and peripherally. κ-opioid agonists are also known to stimulate fat intake. These experiments were conducted to determine if an opioidergic central pathway might mediate the effects of enterostatin and galanin on fat intake. Male Sprague-Dawley rats were adapted to a high-fat diet (56% energy) and were implanted with cannulae aimed at the lateral cerebral ventricle (LV) or third cerebral ventricle (3V). Injection of enterostatin (1 nmol, LV) suppressed high-fat diet consumption in fasted (20 h) rats. This inhibition of high-fat intake by enterostatin was attenuated by central injection of the specific κ-agonist U50488 (2.15, 21.5 and 215 nmol, LV) in a dose-dependent manner in fasted rats while only the highest dose of U50488 (215 nmol, LV) independently produced stimulation of high-fat diet consumption in sated rats. Galanin (0.1 nmol, 3V) induced consumption of high-fat diet in sated rats similar to that seen with U50488 and this stimulation was attenuated by peripheral injection of naloxone (1.0 mg/kg i.p.). We present a model which integrates the present data, as well as previous findings, in explaining a potential common opioid pathway modulating fat consumption.     

Chronic social stress increases levels of preprogalanin mRNA in the rat locus coeruleus

Galanin is a 29 amino acid neuropeptide that coexists with norepinephrine in approximately 80% of locus coeruleus (LC) neurons in the rat. The effects of chronic, naturalistic stress on preprogalanin mRNA in the LC was studied. The visible burrow system (VBS) served as the stress paradigm. Long-Evans rats (three males and two females) were housed together in the VBS for 13 days. The males established dominance hierarchies during this period. On day 14, preprogalanin mRNA in the LC was significantly elevated in subordinate rats compared to dominant and control rats, as measured by quantitative in situ hybridization. Levels of mRNA were positively correlated with the number of wounds by day 7 and day 14 and negatively correlated with body weight gain by day 14. These results suggest that the neuropeptide galanin may be overexpressed during chronic stress in rats.     

Effects of galanin on the secretory activity of the rat adrenal cortex: in vivo and in vitro studies

Galanin, a 28-amino-acid peptide originally isolated from pig intestine, was found to cause dose-dependent rises in the plasma concentration of corticosterone in hypophysectomized rats and in both basal and submaximally ACTH-stimulated in vitro corticosterone production by adrenal quarters and isolated zona fasciculata/reticularis cells. These findings indicate that galanin exerts a direct glucocorticoid secretagogue effect on the inner adrenocortical zones and suggest that galanin, high concentrations of which are contained in adrenal chromaffin cells, may be included in that group of regulatory peptides, by way of which adrenal zona medullaris is thought to exert a paracrine control on the function of the zona corticalis. Galanin was also found to increase the plasma level of aldosterone in hypophysectomized rats in a dose-dependent manner without inducing changes in natremia, kalemia, plasma renin activity, or ACTH plasma concentration. Galanin did enhance both basal aldosterone output and that following submaximal ACTH or angiotesin II stimulation aldosterone out-put from adrenal quarters, but not from isolated zona glomerulosa cells. This last, rather unexpected, result suggests that galanin exerts an indirect mineralocorticoid secretagogue action, which seems to require the structural integrity of the adrenal cortex and the presence of the adrenal medulla. The hypothesis is advanced that galanin may control the release of some medullary peptides, which in turn may affect mineralocorticoid secretion of the zona glomerulosa in a paracrine manner.     

Galantide stimulates sexual behavior in male rats

While intracerebroventricular injection of galanin (5 μg/rat) inhibited sexual behavior in experienced male rats - without producing any other locomotor or behavioral deficit -, injection of the galanin antagonist, galantide, by the same route (1 or 2 μg/rat) stimulated sexual behavior (improving arousal, motivation and performance indexes) and antagonized the effect of galanin. These data further suggest that galanin plays a physiological role in male sexual behavior.     

Neuronal colocalization of adenosine deaminase, monoamine oxidase, galanin and 5-hydroxytryptophan uptake in the tuberomammillary nucleus of the rat

Neurons of the tuberomammillary nucleus (TM) in the rat have previously been shown to contain the enzymes adenosine deaminase (ADA), histidine decarboxylase (HDC) and glutamate decarboxylase (GAD). Some neurons coextensive with this cell group also exhibit immunoreactivity for the neuropeptide galanin, express monoamine oxidase activity (MAO), or display the ability to accumulate and decarboxylate 5-hydroxytryptophan (5-HTP). Histochemical and immunohistochemical techniques were used to determine the extent to which these neurochemical properties are colocalized in neurons immunoreactive for adenosine deaminase. Galanin was found to coexist with ADA in about 45% of the neurons in the TM. In addition, a large number of cells immunoreactive for galanin alone were observed in the posterior hypothalamus outside the confines of TM. Neurons displaying MAO activity formed a subpopulation of those immunoreactive for ADA; all neurons containing MAO also contained ADA whereas only 60% of the ADA-immunoreactive cells were reactive for MAO. Approximately 20% of ADA-immunoreactive neurons represented nearly all cells having 5-HTP uptake capability. However, a very few cells in TM showing 5-HTP uptake capability appeared to be devoid of ADA immunoreactivity. These results demonstrate that although neurons of TM are homogeneous with respect to a number of possible neurotransmitters markers and associated enzymes, these neurons are heterogeneous with respect to their expression of galanin, MAO and 5-HTP uptake. In certain respects the segregation of histochemical properties within TM correlates with previous histochemical work by others, and suggests the possibility of functional diversity of TM.