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The effect of metabolic programming by caloric restriction in areas of central nervous system of Wistar rats involved with feeding behavior: the solitary tract nucleus and the lateral hypothalamic area

Grant number: 16/13136-6
Support type:Regular Research Grants
Duration: December 01, 2016 - November 30, 2018
Field of knowledge:Biological Sciences - Morphology - Anatomy
Principal Investigator:José de Anchieta de Castro e Horta Júnior
Grantee:José de Anchieta de Castro e Horta Júnior
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Maternal nutritional status during critical periods of offspring development (fetal and neonatal), is considered an important Programming inductor, since the nutrient deficiency during pregnancy and lactation, causes significant changes in body weight, energy balance, food intake and expression of neuropeptides. In the central nervous system, these changes occur, particularly in areas that develop during embryonic and neonatal periods and are important to the control of feeding behavior, such as the nucleus of the solitary tract (NTS) which plays an essential role in integration of visceral information related to the control of satiety and the lateral hypothalamic area (LHA) involved with regulation of energy balance and control of food intake. The NTS is a complex consisting of 12 subnuclei where the satiety related signaling is mediated largely by the glutamatergic primary afferents from stomach and duodenum, immunoreactive to vesicular glutamate transporter - type 2 (VGLUT2) and modulated by various neuropeptides. The melanin-concentrating hormone (MCH) and the cocaine and amphetamine regulated transcript (CART) are neuropeptides widely expressed in the central nervous system, especially in the LHA but also in the NTS, in which the MCH plays orexigenic role and CART, anorexigenic. So, our aim is to analyze the expression of MCH and CART and the ultrastructure of immunoreactive neurons to these neuropeptides (MCH-ir and CART-ir) in LHA, to compare the cytoarchitectonic organization and map the immunorreactivity to MCH, CART and VGLUT2 in the NTS of rats whose mothers were subjected to caloric restriction during pregnancy and lactation. For its implementation, mating of adult rats of the Wistar strain will be realized to obtain the offsprings, which are object of study. After detection of pregnancy, females will be separated to form two groups: control group (CG), normal diet ad libitum and caloric restriction group (RG), 50% restriction diet compared to the control group during the periods of pregnancy and lactation. After birth, number of pups and body weight in postnatal day 1 will be evaluated. The male pups, from the restriction (RG) and control (CG) groups, will be divided into five age subgroups (n = 8), 21, 28, 50, 90 and 540 days. In each animal will be evaluated: the body weight and length, food intake after weaning and blood levels of glucose, insulin and leptin. All animals will be transcardially perfused with fixative, the visceral and retroperitoneal adipose tissues and the brain will be weighted. The brains will be processed for analysis of MCH and CART expression in the LHA, according immunohistochemistry, situ hybridization (n=5) and electron microscopy transmission (n=3) protocols. The mapping and estimate of the number of neurons MCH-ir and CART-ir in the LHA will be performed using three-dimensional reconstruction and stereological study respectively. The comparative analysis of the quantity of MCH and CART mRNA in the different experimental groups will be performed by measuring the integrated optical density of labeled area by in situ hybridization. The ultrastructure of neurons MCH-ir and CART-ir will be analyzed by transmission electron microscopy. In the NTS, the distribution of MCH-ir and VGLUT2-ir fibers and CART-ir fibers and somata will be mapped though immunohistochemistry detection followed by optical densitometry and experiments of colocalization by immunofluorescence in the fibers immunoreactive to VGLUT2 and MCH or CART. It will be studied the cytoarchitectonic organization of NTS using the immunoreactivity to calbindin D-28K and nitric oxide synthase as markers, followed by three-dimensional reconstruction. The quantitative data obtained will be compared and statistically analyzed (p <0.05). (AU)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
EMILIO-SILVA, MAYCON T.; RODRIGUES, VINICIUS P.; BUENO, GABRIELA; OHARA, RIE; MARTINS, MARINA G.; HORTA-JUNIOR, JOSE A. C.; BRANCO, LUIZ G. S.; ROCHA, LUCIA R. M.; HIRUMA-LIMA, CLELIA A. Hypothermic Effect of Acute Citral Treatment during LPS-induced Systemic Inflammation in Obese Mice: Reduction of Serum TNF-alpha and Leptin Levels. BIOMOLECULES, v. 10, n. 10 OCT 2020. Web of Science Citations: 0.
PEREIRA, LAIS DA SILVA; GOBBO, DENISE RIBEIRO; FERREIRA, JOZELIA GOMES PACHECO; HORTA-JUNIOR, JOSE DE ANCHIETA DE CASTRO E; SA, SUSANA ISABEL; BITTENCOURT, JACKSON CIONI. Effects of ovariectomy on inputs from the medial preoptic area to the ventromedial nucleus of the hypothalamus of young adult rats. Journal of Anatomy, v. 238, n. 2 SEP 2020. Web of Science Citations: 0.
CABRAL RODRIGUES, LAISA TAIS; SALATA, BRUNO; HORTA-JUNIOR, JOSE DE ANCHIETA C.; GARGAGLIONI, LUCIANE H.; DIAS, MIRELA BARROS. Adenosine in the lateral hypothalamus/perifornical area does not participate on the CO2 chemoreflex. Respiratory Physiology & Neurobiology, v. 276, MAY 2020. Web of Science Citations: 0.

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