Respiratory deficit in an experimental model of Parkinson's Disease may be associated with changes in the Phox2b gene at the level of the ventrolateral medulla

• Grant number: 17/11165-1
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: July 31, 2017
• End date: June 14, 2018
• Field of knowledge: Biological Sciences - Pharmacology - General Pharmacology
• Principal Investigator: Ana Carolina Takakura Moreira
• Grantee: Silvio de Araujo Fernandes Junior
• Supervisor: José Javier Otero
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: Ohio State University, Columbus, United States
• Associated to the scholarship: 14/20695-6 - Analysis of the selective activation of medullary neurons in an experimental model of Parkinson's Disease: a time study of the possible mechanisms for restoration of respiratory function, BP.DR

Abstract

Parkinson's disease (PD) is a disorder characterized by motor and non-motor symptoms, such as respiratory deficits and is associated with loss of dopaminergic neurons in the compact part of the substantia nigra (SNc). However, other brainstem neurons may also be degenerated, contributing to the appearance of non-motor abnormalities. Previous study showed a decrease in the number of phox2b-expressing neurons of retrotrapezoid nucleus (RTN) and a reduction in resting and hypercapnia-induced respiratory frequency. Objective: compare the neuroanatomical and biomolecular changes in the RTN and investigate whether respiratory functional deficit following induction of a PD model may occur by biomolecular changes or neuronal death in cells expressing phox2b nuclear transcription factor within the RTN. Methods: Adult male rats with bilateral injections of 6-hydroxy-dopamine (6-OHDA, 24 µg/µl) or vehicle into the striatum will be used. Brainstem will be cut into 8 ¼m sections and kept at 80°C until use. Laser capture and pressure catapulting (LMPC) will be performed using the laser microdissection system from PALM Technologies (Carl Zeiss MicroImaging GmbH, Munchen, Germany). Total number of cells captured will be determined using the conversion factor of 50,000 ¼m2. Upon completion of microdissection, the captured material will be spun down into a 0.2 ml PCR tube and held at -80°C until protein retrieval. For mass spectrometry, peptides will be separated by reversed-phase HPLC (Dionex Ultimate 3000 capillary/nano HPLC system, Dionex, Sunnyvale, CA) and mass analyzed with a Thermo Fisher LTQ Orbitrap XL, equipped with micro/nanospray ionization sources (Michrom Bioresources Inc., Auburn, CA). Immunohistochemistry will be performed for the detection of the tyrosine-hydroxylase and evaluate the extent and selectivity of the 6-OHDA lesion. (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)

FERNANDES-JUNIOR, SILVIO A.; OLIVEIRA, LUIZ M.; CZEISLER, CATHERINE M.; MO, XIAOKUI; ROY, SASHWATI; SOMOGYI, ARPAD; ZHANG, LIEWN; MOREIRA, THIAGO S.; OTERO, JOSE J.; TAKAKURA, ANA C.. Stimulation of retrotrapezoid nucleus Phox2b-expressing neurons rescues breathing dysfunction in an experimental Parkinson's disease rat model. Brain Pathology, v. 30, n. 5, JUL 2020. (16/23281-3, 14/20695-6, 15/23376-1, 17/11165-1)

OLIVEIRA, LUIZ M.; FERNANDES-JUNIOR, SILVIO A.; CABRAL, LAIS M. C.; MIRANDA, NICOLE C. S.; CZEISLER, CATHERINE M.; OTERO, JOSE J.; MOREIRA, THIAGO S.; TAKAKURA, ANA C.. Regulation of blood vessels by ATP in the ventral medullary surface in a rat model of Parkinson's disease. Brain Research Bulletin, v. 187, p. 17-pg., 2022-07-12. (15/23376-1, 17/11165-1, 19/09323-3, 14/20695-6, 19/01236-4)