Immunohistochemical Evidence for Glutamatergic Regulation of Nesfatin-1 Neurons in the Rat Hypothalamus

Nesfatin-1, identified as an anorexigenic peptide, regulates the energy metabolism by suppressing food intake. The majority of nesfatin-1-synthesizing neurons are concentrated in various hypothalamic nuclei, especially in the supraoptic (SON), arcuate (ARC) and paraventricular nuclei (PVN). We tested the hypothesis that the glutamatergic system regulates nesfatin-1 neurons through glutamate receptors. Therefore, the first aim of the proposed studies was to examine effects of different glutamate agonists in the activation of nesfatin-1 neurons using c-Fos double immunohistochemical labeling. Experimental groups were formed containing male and female rats which received intraperitoneal injections of glutamate agonists kainic acid, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA) while the control rats received vehicle. The significant increase in the number of c-Fos-expressing nesfatin-1 neurons after agonist injections were observed both in female and male subjects and some of these effects were found to be sexually dimorphic. In addition, treatment with specific glutamate antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) or dizocilpine (MK-801) before each of the three agonist injections caused a statistically significant reduction in the number of activated nesfatin-1 neurons in the hypothalamic nuclei including supraoptic, paraventricular and arcuate nuclei. The second aim of the study was to determine the expression of glutamate receptor subunit proteins in the nesfatin-1 neurons by using a double immunofluorescence technique. The results showed that the glutamate receptor subunits, which may form homomeric or heteromeric functional receptor channels, were expressed in the nesfatin-1 neurons. In conclusion, the results of this study suggest that nesfatin-1 neurons respond to glutamatergic signals in the form of neuronal activation and that the glutamate receptors that are synthesized by nesfatin-1 neurons may participate in the glutamatergic regulation of these neurons.

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Proteins Regulated by Gonadal Steroids in the Medial Preoptic and Ventromedial Hypothalamic Nuclei of Male and Female Rats

Neuroendocrinology ◽

10.1159/000124183 ◽

1985 ◽

Vol 41 (3) ◽

pp. 237-245 ◽

Cited By ~ 30

Author(s):

Charles W. Scouten ◽

William E. Heydorn ◽

G. Joseph Creed ◽

Charles W. Malsbury ◽

David M. Jacobowitz

Keyword(s):

Gonadal Steroids ◽

Female Rats ◽

Male And Female ◽

Male And Female Rats ◽

Hypothalamic Nuclei

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A peripheral neuroimmune link: glutamate agonists upregulate NMDA NR1 receptor mRNA and protein, vimentin, TNF-α, and RANTES in cultured human synoviocytes

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00452.2009 ◽

2010 ◽

Vol 298 (3) ◽

pp. R584-R598 ◽

Cited By ~ 22

Author(s):

Terry A. McNearney ◽

Yinghong Ma ◽

Yueping Chen ◽

Giulio Taglialatela ◽

Huaizhi Yin ◽

...

Keyword(s):

Protein Kinase ◽

Glutamate Receptors ◽

Glutamate Receptor ◽

Immunocytochemical Staining ◽

Target Cells ◽

Synovial Cells ◽

Blocking Antibody ◽

Proinflammatory Mediators ◽

Tnf Α ◽

Glutamate Agonists

Human primary and clonal synovial cells were incubated with glutamate receptor agonists to assess their modulating influence on glutamate receptors N-methyl-d-aspartate (NMDA) NR1 and NR2 and inflammatory cytokines to determine potential for paracrine or autocrine (neurocrine) upregulation of glutamate receptors, as has been shown for bone and chondrocytes. Clonal SW982 synoviocytes constitutively express vimentin, smooth muscle actin (SMA), and NMDA NR1 and NR2. Coincubation (6 h) with glutamate agonists NMDA (5 μM), and the NMDA NR1 glycine site activator (±)1-aminocyclopentane- cis-1,3-dicarboxylic acid (5 μM), significantly increases cellular mRNA and protein levels of glutamate receptors, as well as increasing vimentin, SMA, tumor necrosis factor-α, and RANTES (regulated on activation, normal T-cell expressed and secreted), assessed qualitatively and quantitatively with nucleotide amplification, image analysis of immunocytochemical staining, fluorescein-activated cell sorting, Western blotting, and immunoassays. Human primary synovial cells harvested from patients with arthritic conditions also constitutively expressed NMDA NR1 with increases after agonist treatment. Glutamate receptor agonist-induced increases were blocked by the noncompetitive glutamate antagonist MK-801 (8 μg/ml) and NR1 blocking antibody. Coincubation with glutamate agonists and phorbol 12-myristate 13-acetate, a protein kinase C activator, significantly enhanced mean levels of TNF-α and RANTES in SW982 cell supernatants compared with incubation with either agent alone. Increases were diminished with protein kinase inhibitor and NR1 blocking antibody. The functional activation of glutamate receptors on human synoviocytes establishes a neurogenic cell signaling link between neurotransmitter glutamate released from nerve terminals and target cells in the joint capsule. The influence of glutamate on subsequent release of cellular proinflammatory mediators in non-neural tissue for activation of downstream immune events supports a peripheral neuroimmune link in arthritis.

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The role of glutamate and nitric oxide in the reproductive neuroendocrine system

Biochemistry and Cell Biology ◽

10.1139/o00-015 ◽

2000 ◽

Vol 78 (3) ◽

pp. 165-179 ◽

Cited By ~ 35

Author(s):

Krishnan M Dhandapani ◽

Darrell W Brann

Keyword(s):

Nitric Oxide ◽

Glutamate Receptors ◽

Recent Work ◽

Hypothalamic Nuclei ◽

New Findings ◽

Gnrh Release ◽

Glutamate Agonists ◽

Lh Release ◽

Reproductive Events

The preovulatory surge of gonadotropin releasing hormone (GnRH) is essential for mammalian reproduction. Recent work has implicated the neurotransmitters glutamate and nitric oxide as having a key role in this process. Large concentrations of glutamate are found in several hypothalamic nuclei known to be important for GnRH release and glutamate receptors are also located in these key hypothalamic nuclei. Administration of glutamate agonists stimulate GnRH and LH release, while glutamate receptor antagonists attenuate the steroid-induced and preovulatory LH surge. Glutamate has also been implicated in the critical processes of puberty, hormone pulsatility, and sexual behavior. Glutamate is believed to elicit many of these effects by activating the release of the gaseous neurotransmitter, nitric oxide (NO). NO potently stimulates GnRH by activating a heme containing enzyme, guanylate cyclase, which in turn leads to increased production of cGMP and GnRH release. Recent work has focused on identifying anchoring and (or) clustering proteins that target glutamate receptors to the synapse and couple the glutamate-NO neurotransmission system. The present review will discuss these new findings, as well as the role of glutamate and nitric oxide in important mammalian reproductive events, with a focus on the hypothalamic control of preovulatory GnRH release. Key words: glutamate, nitric oxide, GnRH, postsynaptic density, hypothalamus.

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Monosodium glutamate administration early in life alters pineal melatonin nocturnal profile in adulthood

Melatonin Research ◽

10.32794/mr11250084 ◽

2021 ◽

Vol 4 (1) ◽

pp. 99-114

Author(s):

Janaína B Garcia ◽

Fernanda G Do Amaral ◽

Daniela C Buonfiglio ◽

Rafaela FA Vendrame ◽

Patrícia L Alves ◽

...

Keyword(s):

Insulin Resistance ◽

Body Weight ◽

Pineal Gland ◽

Serum Insulin ◽

Monosodium Glutamate ◽

Female Rats ◽

Pineal Melatonin ◽

Melatonin Synthesis ◽

Male And Female Rats ◽

Melatonin Production

The pineal gland synthesizes melatonin exclusively at night, which gives melatonin the characteristic of a temporal synchronizer of the physiological systems. Melatonin is a regulator of insulin activities centrally and also peripherally and its synthesis is reduced in diabetes. Since monosodium glutamate (MSG) is often used to induce the type 2 diabetic and metabolic syndrome in animal models, the purpose of this work is to evaluate the potential effects of MSG given to neonates on the pineal melatonin synthesis in different aged male and female rats. Wistar rats were subcutaneously injected with MSG (4mg/g/day) or saline solution (0.9%) from the second to eighth post-natal day. The circadian profiles both melatonin levels and AANAT activity were monitored at different ages. Body weight, naso-anal length, adipose tissues weight, GTT, ITT and serum insulin levels were also evaluated. Typical obesity with the neonatal MSG treatment was observed, indicated by a great increase in adipose depots without a concurrent increase in body weight. MSG treatment did not cause hyperglycemia or glucose intolerance, but induced insulin resistance. An increase of melatonin synthesis at ZT 15 with phase advance was observed in in some animals. The AANAT activity was positively parallel to the melatonin circadian profile. It seems that MSG causes hypothalamic obesity which may increase AANAT activity and melatonin production in pineal gland. These effects were not temporally correlated with insulin resistance and hyperinsulinemia indicating the hypothalamic lesions, particularly in arcuate nucleus induced by MSG in early age, as the principal cause of the increase in melatonin production.

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THE PITUITARY AND ADRENAL RESPONSES TO ADMINISTRATION OF OESTRIOL IN GONADECTOMIZED RATS

Acta Endocrinologica ◽

10.1530/acta.0.0380050 ◽

1961 ◽

Vol 38 (1) ◽

pp. 50-58 ◽

Cited By ~ 3

Author(s):

N. E. Borglin ◽

L. Bjersing

Keyword(s):

Pituitary Gland ◽

Adrenal Cortex ◽

Clinical Experience ◽

Uterine Cervix ◽

Morphological Changes ◽

Physiological Significance ◽

Female Rats ◽

Experimental Conditions ◽

Male And Female ◽

Male And Female Rats

ABSTRACT Oestriol (oestra-1,3,5(10)-triene-3,16α,17β-triol) is a weakly oestrogenic substance which, however, in contrast to what was formerly believed, is of physiological significance. Its effect is localized largely to the uterine cervix and vagina. Clinical experience argues both for and against an effect on the pituitary gland. This investigation is concerned with the morphological changes in the pituitary gland and adrenal cortex of gonadectomized male and female rats after the injection of oestriol. It was found that oestriol has the same type of action on these glands as other oestrogens, but under the experimental conditions used, this effect proved much weaker than that produced by oestradiol (oestra-1,3,5(10)-triene-3,17β-diol).

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INDUCTION OF GOITRE BY PTU OR KClO4 IN MALE AND FEMALE RATS. EFFECT OF GONADECTOMY

Acta Endocrinologica ◽

10.1530/acta.0.0740088 ◽

1973 ◽

Vol 74 (1) ◽

pp. 88-104 ◽

Cited By ~ 2

Author(s):

T. Jolín ◽

M. J. Tarin ◽

M. D. Garcia

Keyword(s):

Iodine Content ◽

High Plasma ◽

Disc Electrophoresis ◽

Female Rats ◽

Male Rats ◽

Adult Rats ◽

Male And Female ◽

Glucose Levels ◽

Male And Female Rats ◽

Tsh Levels

ABSTRACT Male and female rats of varying ages were placad on a low iodine diet (LID) plus KClO4 or 6-propyl-2-thiouracil (PTU) or on the same diet supplemented with I (control rats). Goitrogenesis was also induced with LID plus PTU in gonadectomized animals of both sexes. The weight of the control and goitrogen treated animals, and the weight and iodine content of their thyroids were determined, as well as the plasma PBI, TSH, insulin and glucose levels. The pituitary GH-like protein content was assessed by disc electrophoresis on polyacrylamide gels. If goitrogenesis was induced in young rats of both sexes starting with rats of the same age, body weight (B.W.) and pituitary growth hormone (GH) content, it was found that both the males and females developed goitres of the same size. On the contrary, when goitrogenesis was induced in adult animals, it was found that male rats, that had larger B.W. and pituitary GH content than age-paired females, developed larger goitres. However, both male and female rats were in a hypothyroid condition of comparable degree as judged by the thyroidal iodine content and the plasma PBI and TSH levels. When all the data on the PTU or KClO4-treated male and female rats of varying age and B.W. were considered together, it was observed that the weights of the thyroids increased proportionally to B.W. However, a difference in the slope of the regression of the thyroid weight over B.W. was found between male and female rats, due to the fact that adult male rats develop larger goitres than female animals. In addition, in the male rats treated with PTU, gonadectomy decreased the B.W., pituitary content of GH-like protein and, concomitantly, the size of the goitre decreased; an opposite effect was induced by ovariectomy on the female animals. However, when goitrogenesis was induced in weight-paired adult rats of both sexes, the male animals still developed larger goitres than the females. Among all the parameters studied here, the only ones which appeared to bear a consistent relationship with the size of the goitres in rats of different sexes, treated with a given goitrogen, were the rate of body growth and the amount of a pituitary GH-like protein found before the onset of the goitrogen treatment. Moreover, though the pituitary content of the GH-like protein decreased as a consequence of goitrogen treatment, it was still somewhat higher in male that in female animals. The present results suggest that GH may somehow be involved in the mechanism by which male and female rats on goitrogens develop goitres of different sizes, despite equally high plasma TSH levels.

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THE UPTAKE AND LOCALIZATION OF 3H-OESTRADIOL IN THE HYPOTHALAMUS OF MALE AND FEMALE RATS

Acta Endocrinologica ◽

10.1530/acta.0.061s194 ◽

1969 ◽

Vol 61 (1_Suppl) ◽

pp. S194 ◽

Cited By ~ 1

Author(s):

Arne Attramadal

Keyword(s):

Female Rats ◽

Male And Female ◽

Male And Female Rats

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GONADOTROPHIN PATTERNS IN MALE AND FEMALE RATS:

Acta Endocrinologica ◽

10.1530/acta.0.0580600 ◽

1968 ◽

Vol 58 (4) ◽

pp. 600-612 ◽

Cited By ~ 1

Author(s):

Robert Boyd ◽

Donald C. Johnson

Keyword(s):

Ascorbic Acid ◽

Testosterone Propionate ◽

Female Rats ◽

Female Rat ◽

Feedback Effects ◽

Male And Female ◽

Prostate Weight ◽

Male And Female Rats ◽

Males And Females ◽

Normal Males

ABSTRACT The effects of various doses of testosterone propionate (TP) upon the release of luteinizing hormone (LH or ICSH) from the hypophysis of a gonadectomized male or female rat were compared. Prostate weight in hypophysectomized male parabiotic partners was used to evaluate the quantity of circulating LH. Hypophyseal LH was measured by the ovarian ascorbic acid depletion method. Males castrated when 45 days old secreted significantly more LH and had three times the amount of pituitary LH as ovariectomized females. Administration of 25 μg TP daily reduced the amount of LH in the plasma, and increased the amount in the pituitary gland, in both sexes. Treatment with 50 μg caused a further reduction in plasma LH in males, but not in females, while pituitary levels in both were equal to that of their respective controls. LH fell to the same low level in partners of males or females receiving 100 μg TP. When gonadectomized at 39 days, males and females had the same amount of plasma LH, but males had more stored hormone. Pituitary levels were unchanged from controls following treatment with 12.5, 25 or 50 μg TP daily, but plasma values dropped an equal amount in both sexes with the latter two doses. Androgenized males or females, gonadectomized when 39 days old, were very sensitive to the effects of TP and plasma LH was significantly reduced with 12.5 μg daily. Pituitary LH in androgenized males was higher than that of normal males but was reduced to normal by small amounts of TP. The amount of stored LH in androgenized females was not different from that of normal females and it was unchanged by any dose of TP tested. Results are consistent with the conclusion that the male hypothalamic-hypophyseal axis is at least as sensitive as the female axis to the negative feedback effects of TP. Androgenization increases the sensitivity to TP in both males and females.

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