stress induction methods for establishing the chronic unpredictable mild stress rat model

Etablierung und Validierung eines Modells für chronisch unvorhersehbaren leichten Stress

Major depressive disorder (MDD) is a prevalent mental illness, ranking as the third leading cause of disability globally and affecting over 300 million people1,2,3. Notably, it is estimated that at least half of the affected individuals do not receive adequate treatment4. Given this gap, animal models serve as a crucial tool for investigating the etiology of depression. To date, more than 20 different animal models for depression exist5. Among these, the chronic unpredictable mild stress (CUMS) model, refined by Paul Winer in 1987, is the most frequently utilized6. The CUMS model operates on the premise that exposing rodents to a diverse range of socio-environmental stressors leads to symptoms akin to anxiety, tension, and depression. The methodology involves exposing animals to various mild stressors over several weeks, culminating in a range of behavioral alterations, including anhedonia and depressive-like behaviors7,8. These changes are accompanied by shifts in endocrine and neurotransmitter profiles, such as a reduction in 5-HT9,10. These outcomes closely mirror those observed in humans diagnosed with MDD, thereby validating the model's utility. The CUMS model is particularly valued for its effectiveness in evaluating antidepressants, manifesting high levels of surface, structural, and predictive validity11,12. Unlike other models, CUMS is sensitive to the effects of chronic administration of monoaminergic antidepressants. For instance, selective serotonin reuptake inhibitors (SSRIs) such as citalopram, paroxetine, and fluoxetine have been shown to prevent and reverse anhedonia under conditions of chronic stress12,13. Additionally, new rapid-acting antidepressants, such as ketamine, have also demonstrated efficacy in this model14,15. In contrast, other tests like the forced swimming test (FST) and tail suspension test (TST) are less reliable for modeling long-term behavioral changes, often reflecting adaptations to acute stress rather than enduring symptoms of depression16. These characteristics underscore the robust validity of the CUMS model in depression research. One of the most salient features of the CUMS model, recognized for its high reliability in classical studies, is anhedonia-the incapacity to experience pleasure or interest in everyday activities17,18. This phenomenon is commonly evaluated using sucrose preference tests, and many antidepressants have been shown to reverse reduced sucrose consumption. Several other metrics are also commonly employed in the CUMS literature, including the open field test (OFT), which assesses voluntary motor behavior, exploratory tendencies, and tension, thereby gauging the severity of depression19. Other tests such as the elevated plus maze (EPM) evaluate anxiety-like behaviors, the Morris water maze test (MWM) examines cognitive functioning20, and the FST assesses susceptibility to negative emotions and behavioral despair20. Furthermore, the majority of stressors that impact humans are social in nature. Individuals with suboptimal social relationships, characterized by limited social activities, networks, and support, are at a heightened risk for various diseases21,22. This is also relevant in the case of rodents, who are social animals living in groups. For example, rats that are housed in isolation exhibit characteristics of what is termed isolation syndrome, which induces social stress and expedites the onset of depression23.
Mongolian medicine, a significant branch of Chinese medicine, posits that the onset of disease is a complex interplay between intrinsic and external factors. These external factors, referred to as the four auxiliary conditions, encompass climate change, diet, lifestyle, and sudden events such as infections, startling incidents, and psychological disorders. The disease process is conceptualized as an ongoing interaction between three elements-referred to as the three types of homors-and the Seven Bodily Constituents in concert with the four auxiliary conditions24. Mongolian medicine holds that the human body functions as an integrated entity, maintained by a relative balance among the three homors. A disruption in this balance is considered to be a precursor to illness24. Given the pivotal role of animal experimentation in bridging traditional and modern medicine, it is crucial to develop animal models that are relevant for research in the field of Mongolian medicine. Accordingly, we employed a 28-day isolation methodology coupled with CUMS to simulate these physiological and psychological stressors. We selected nine specific unpredictable stressors and sought to underpin this modeling method through the Three homors theory of Mongolian medicine. Establishing a robust animal model is fundamental for advancing basic research in Mongolian medicine and will contribute significantly to its foundational studies.

Autor im Rampenlicht: Etablierung eines Nagetiermodells zur Untersuchung von Depressionsfaktoren in der traditionellen mongolischen Medizin

Chronisch unvorhersehbarer milder Stress bei Ratten basierend auf der mongolischen Medizin

Stressinduktionsmethoden zur Etablierung des Modells der chronisch unvorhersehbaren leichten Stressratte

Nachdem Sie die Ratten in eine Kontroll-, Betrüger- und Modell- oder Mod-Gruppe unterteilt haben, halten Sie sie in einzelnen Käfigen unter. Sichern Sie für die Schwanzklemmmethode den Schwanz einer Ratte und messen Sie die Klemmzeit für eine Minute. Bei der Wasserentzugsmethode nehmen Sie die Babyflasche aus dem Käfig und nehmen die Ratte 24 Stunden lang auf.Für die Methode des Nahrungsentzugs nehmen Sie das Futter aus dem Käfig und nehmen Sie die Ratte 24 Stunden lang auf. Für eine Kältestimulation bei vier Grad Celsius legen Sie eine Ratte in einen Eimer mit kaltem Wasser und überwachen Sie ihre Aktivität fünf Minuten lang. Trocknen Sie die Ratte dann mit einem Gebläse ab und setzen Sie sie wieder in ihren ursprünglichen Käfig ein.Um 45 Grad Celsius Hitzestress zu bewältigen, stellen Sie die Ratte in einen Ofen und überwachen Sie sie fünf Minuten lang. Für die Umkehrung des Hell-Dunkel-Zyklus wickeln Sie den Käfig eine Stunde lang in ein schwarzes Tuch, um die Dunkelheit am Tag zu stimulieren. Beleuchten Sie den Käfig anschließend nachts 12 Stunden lang, um das Tageslicht nachzuahmen und die Daten 24 Stunden lang aufzuzeichnen.Gießen Sie für das Experiment mit der nassen Polsterung 200 Milliliter Wasser in einen Käfig mit 100 Gramm Polsterung. Setzen Sie die Ratte in den nassen Käfig und überwachen Sie sie 24 Stunden lang. Nachdem Sie die Ratte wie bereits gezeigt getrocknet haben, setzen Sie sie in einen Käfig mit frischen Holzspänen zurück.Setzen Sie die Ratte bei der Käfigneigungsmethode in einen Käfig, der um 45 Grad an einer Wand geneigt ist, und überwachen Sie sie 24 Stunden lang. Setzen Sie die Ratte für ein Schütteln mit hoher Geschwindigkeit in die mechanischen Shaker ein, die auf 160 U/min eingestellt sind, und zeichnen Sie die Daten 15 Minuten lang auf. Übertragen Sie nach dem Anwenden von Stressoren den Mod-Gruppenkäfig aus dem CUMS-Raum in den Haltungsraum.

stress-induction-methods-for-establishing-chronic-unpredictable-mild

Research

JoVE Journal

Medicine

Stress Induction Methods for Establishing the Chronic Unpredictable Mild Stress Rat Model

77 Aufrufe. Nachdem Sie die Ratten in eine Kontroll-, Betrüger- und Modell- oder Mod-Gruppe unterteilt haben, halten Sie sie in einzelnen Käfigen unter. Sichern Sie für die Schwanzklemmmethode den Schwanz einer Ratte und messen Sie die Klemmzeit für eine Minute. Bei der Wasserentzugsmethode nehmen Sie die Babyflasche aus dem Käfig und nehmen die Ratte 24 Stunden lang auf.Für die Methode des Nahrungsentzugs nehmen Sie das Futter aus dem Käfig und nehmen Sie die Ratte 24 Stunden lang auf. Fü...

Serie: Stressinduktionsmethoden zur Etablierung des Modells der chronisch unvorhersehbaren leichten Stressratte

Chronic Unpredictable Mild Stress in Rats based on the Mongolian medicine

Depression is a prevalent affective disorder and constitutes a leading cause of global disability. The limitations of current pharmacological interventions contribute to the substantial health burden attributed to this condition. There is a pressing need for a deeper understanding of the underlying mechanisms of depression, making pre-clinical models with translational potential highly valuable. Mongolian medicine, a subset of traditional medicine, posits that disease occurrence is closely tied to the equilibrium of wind, bile, and Phlegm. In this study, we introduce a protocol for the chronic unpredictable mild stress (CUMS) method in rats. Within this framework, rats are subjected to a series of fluctuating, mild stressors to induce a depression-like phenotype, mimicking the pathogenesis of human depression. Behavioral assays employed in this protocol include the sucrose preference test (SPT), indicative of anhedonia-a core symptom of depression; the open field test (OFT), which measures anxiety levels; and the Morris water maze test (MWM), which evaluates spatial memory and learning abilities. The CUMS method demonstrates the capability to induce anhedonia and to cause long-term behavioral deficits. Furthermore, this protocol is more aligned with Mongolian medical theory than other animal models designed to elicit depression-like behavior. The development of this animal model and subsequent research provide a robust foundation for future innovative studies in the realm of Mongolian medicine.

This protocol outlines a chronic unpredictable mild stress (CUMS) model for depression based on Mongolian medical theory, along with methods for validating behavioral tests.