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INVESTIGATION OF GENES AND PROTEINS RELATED TO RESTLESS LEGS SYNDROME IN ANIMAL MODEL
Restless legs syndrome (RLS) is a neurological disorder that manifests itself as an uncontrollable desire to move, mainly, the legs to relieve uncomfortable sensations. Its pathophysiology is related to dopaminergic dysfunction, reduced iron and variations in gene expression, such as PTPRD (Protein Tyrosine Phosphatase Receptor Type Delta), which can be caused by single nucleotide polymorphisms. In addition, changes in this gene can also be linked to hypertension.
The present study aimed to evaluate the levels of gene transcripts and protein content of genes and proteins of the dopaminergic pathway and PTPRD; in addition to analyzing the sleep pattern, as well as the cardiovascular parameters in an animal model of the Restless Legs Syndrome (Spontaneously Hypertensive Rats-SHR).
For this purpose, the rats were divided into 2 groups: 1) Wistar-Kyoto (standard control); 2) SHR control (SHR CTRL). The animals were submitted to the measurement of cardiovascular parameters of systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate (HR), through tail plethysmography. To assess the sleep pattern, electrocortical and electromyographic analyzes were performed in the 24-hour period (12h light cycle/ 12h dark cycle). For the PTPRD analyses, the qPCR (mRNA levels) and indirect ELISA (protein content) techniques were used. To evaluate the tyrosine-hydroxylase (TH) enzyme, the dopamine transporter (DAT) and the type 2 dopaminergic receptor (DA-2) the qPCR (mRNA levels) and Western Blotting (protein content) techniques were used. For the quantification of iron, ferritin and transferrin the ELISA method was used.
The results showed that SHR animals have higher SBP and DBP, expressed lower levels of protein content of PTPRD, lower levels of gene transcripts and protein content DAT, lower serum concentrations of ferritin, besides changes in sleep pattern (higher number of awakenings, higher percentage of wakefulness, in addition to shorter total time of sleep, sleep efficiency, slow wave sleep and REM sleep), in relation to Wistar-Kyoto animals. Furthermore, SHR animals have higher SBP and DBP.
It is suggested that the molecular alterations observed in SHR animals, as well as in cardiovascular parameters, may be associated with alterations in the sleep pattern, as well as with RLS symptoms in these animals.
Cardiovascular Diseases; Polysomnography; Protein Tyrosine Phosphatase; Sleep- Related Movement Disorder
Faculdade de Ciências Aplicadas - UNICAMP - São Paulo - Brasil
Milca Abda Morais, Beatriz Silva Franco, Alessandro Spencer Souza Holanda, Adriana Souza Torsoni, Andrea Maculano Esteves