Sirtuin 1 Expression in the Rat Ventromedial and Dorsomedial Hypothalamic Nuclei during Ageing
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Bulletin of Experimental Biology and Medicine, Vol. 169, No. 5, September, 2020
BIOGERONTOLOGY Sirtuin 1 Expression in the Rat Ventromedial and Dorsomedial Hypothalamic Nuclei during Ageing A. A. Spirichev, K. Yu. Moiseev, and P. M. Masliukov
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 169, No. 5, pp. 631-633, May, 2020 Original article submitted December 6, 2019 We studied the expression of sirtuin 1 (Sirt1) in the dorsomedial and ventromedial nuclei of the hypothalamus in young (2 months) and old (2 years) rats by immunohistochemical methods and Western blotting. In aged males and females, a decrease in Sirt1 expression in dorsomedial nucleus was observed. In ventromedial nucleus, the expression of Sirt1 did not change with age. The results confirm the hypothesis that aging is associated with a decrease in the content of sirtuins in the hypothalamic nuclei. Key Words: sirtuins; hypothalamus; immunohistochemistry; Western blotting; aging
The hypothalamus plays an important role in homeostasis maintenance and integration of the autonomic, endocrine, and somatic systems. The hypothalamus is also involved in aging mechanisms [3,6,8,11]; an important role is played by the middle group of hypothalamic nuclei, including the ventromedial (VMH) and dorsomedial (DMH) [10]. VMH and DMH are involved in the regulation of trophic processes and maintenance of the energy homeostasis, as well as in the control of peripheral circadian rhythms [7,8]. The expression of calcium-binding proteins and neuronal NO synthase in VMH and DMH changes during aging [1]. Sirtuins (Silent Information Regulator 2 (Sir2) proteins) are a class of proteins with the properties of histone deacetylase and monoribosyl transferase [2]. In many cells, sirtuins interact with transcriptional factors involved in ageing control, such as FOXO and target of rapamycin (mTOR) [6]. An important role in the regulation of ageing and longevity in mammals is played by sirtuin 1 (Sirt1) [6]. However, changes in Yaroslalv State Medical University, Ministry of Health of the Russian Federation, Yaroslavl, Russia. Address for correspondence: mpm@ ysmu.ru. P. M. Masliukov 1
Sirt1 expression in hypothalamic nuclei during ageing remain poorly studied. Here we studied the expression of Sirt1 in VMH and DMH of the hypothalamus during ageing by immunohistochemical methods and by Western blotting.
MATERIALS AND METHODS The study was carried out on 20 white Wistar male and female rats at the age of 2 months (young) and 2 years (aged) in compliance with the Rules for Carrying out Work using Experimental Animals. DMH and VMH nuclei were analyzed. After intraperitoneal injection of a lethal dose of urethane (3 g/kg), the animals were transcardially perfused with 0.01 M PBS (pH 7.4; BioloT). For immunohistochemical studies, the animals were perfused with 4% paraformaldehyde solution (Sigma) in PBS. Then, the brain was removed and the hypothalamic region containing DMH and VMH was extracted according to the coordinates of rat brain atlas [4]. Corona
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