Altered ratio of proapoptotic and antiapoptotic proteins in different brain regions of female rats in model of post-traumatic stress disorder

Abstract

Objective: The B-cell lymphoma/leukemia-2 (Bcl-2) family of proteins governs mitochondrial membrane permeability where the programmed apoptotic process is controlled by the balance between proapoptotic (Bax) and antiapoptotic (Bcl-2) proteins. We aimed to investigate the [Bcl-2]/[Bax] in different brain regions in a post-traumatic stress disorder rat model. Methods: Female Sprague-Dawley rats were exposed to dirty cat litter (trauma) for 10 min and the protocol was repeated 1 week later with a trauma reminder (clean litter) in reversed 12 h light/dark cycle. The rats received intraperitoneal saline, fluoxetine (2.5 mg/kg/day) or propranolol (10 mg/kg/day) for 7 days between exposure sessions. Following exposure to the trauma reminder, elevated plus maze experiments were done. Immunoblotting was used to quantify and [Bax] proteins in the homogenates of the dorsal hippocampus, the frontal cortex and the amygdaloid complex. pathology of the disease [16] and our previous data indicated that the cat litter test is a reproducible experiment where the rats subjected to dirty cat litter (the trauma) had longer freezing times and higher anxiety indices when exposed to the situational reminder, the clean cat litter [17]. Our previous another data showed that noradrenaline (NA) content in the rostral pons increased in traumatized rats with cat litter and this increase may serve as an additional neurochemical evidence for the model. Pirenzepine suppressed the anxiety index but it was not that effective in decreasing the prolonged freezing times [18].In this current study, we wanted to demonstrate the ratio between proapoptotic and antiapoptotic proteins in the dorsal hippocampus, the amygdaloid complex and the frontal cortex of female rats traumatized with cat litter. The effects of fluoxetine or propranolol on behavioral parameters and neuronal apoptosis were also investigated.

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“…In the next stage, they were incubated for another hour with a secondary antibody containing alkaline phosphatase conjugated to rabbit monoclonal anti-goat immunoglobulin G. After the secondary antibody incubation, membranes were washed once more using TBS-T for 3x15 min. Following the secondary antibody incubation, distinctive bands were identified on the membranes after being washed using TBS-T for 3×15 min one more time and usingn blue tetrazolium chloride/5-Bromo-4chloro-3indolyl phosphate color improvement solution (18,19).…”

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“…Apoptotic induction induced movement of Bax from cytosol to mitochondria, leading to multimerization, integration and cytochrome c release, caspase activation, and apoptotic cell death (Aykaç et al 2015). While Bcl-2 defends cells from apoptotic cell death, the over expression of Bax can stimulate apoptosis (Salim et al 2014).…”