Effects of the Selective Serotonin Reuptake Inhibitor Fluoxetine on Developing Neural Circuits in a Model of the Human Fetal Cortex

The developing prenatal brain is particularly susceptible to environmental disturbances. During prenatal brain development, synapses form between neurons, resulting in neural circuits that support complex cognitive functions. In utero exposure to environmental factors such as pharmaceuticals that alter the process of synapse formation increases the risk of neurodevelopmental abnormalities. However, there is a lack of research into how specific environmental factors directly impact the developing neural circuitry of the human brain. For example, selective serotonin reuptake inhibitors are commonly used throughout pregnancy to treat depression, yet their impact on the developing fetal brain remains unclear. Recently, human brain models have provided unprecedented access to the critical window of prenatal brain development. In the present study, we used human neurons and cortical spheroids to determine whether the selective serotonin reuptake inhibitor fluoxetine alters neurite and synapse formation and the development of spontaneous activity within neural circuits. We demonstrate that cortical spheroids express serotonin transporter, thus recapitulating the early developmental expression of serotonin transporter associated with cortical pyramidal neurons. Cortical spheroids also appropriately express serotonin receptors, such as synaptic 5-HT2A and glial 5-HT5A. To determine whether fluoxetine can affect developing neural circuits independent of serotonergic innervation from the dorsal and medial raphe nuclei, we treated cortical neurons and spheroids with fluoxetine. Fluoxetine alters neurite formation in a dose-dependent fashion. Intriguingly, in cortical spheroids, neither acute nor chronic fluoxetine significantly altered excitatory synapse formation. However, only acute, but not chronic fluoxetine exposure altered inhibitory synaptogenesis. Finally, fluoxetine reversibly suppresses neuronal activity in a dose-dependent manner. These results demonstrate that fluoxetine can acutely alter synaptic function in developing neural circuits, but the effects were not long-lasting. This work provides a foundation for future studies to combine serotonergic innervation with cortical spheroids and assess the contributions of fluoxetine-induced alterations in serotonin levels to brain development.

Information capacity and robustness of encoding in the medial prefrontal cortex are modulated by the bioavailability of serotonin and the time elapsed from the cue during a reward-driven task

Serotonin (5-HT) is a key neuromodulator of medial prefrontal cortex (mPFC) functions. Pharmacological manipulation of systemic 5-HT bioavailability alters the electrical activity of mPFC neurons. However, 5-HT modulation at the population level is not well characterized. In the present study, we made single neuron extracellular recordings in the mPFC of rats performing an operant conditioning task, and analyzed the effect of systemic administration of fluoxetine (a selective serotonin reuptake inhibitor) on the information encoded in the firing activity of the neural population. Chronic (longer than 15 days), but not acute (less than 15 days), fluoxetine administration reduced the firing rate of mPFC neurons. Moreover, fluoxetine treatment enhanced pairwise entropy but diminished noise correlation and redundancy in the information encoded, thus showing how mPFC differentially encodes information as a function of 5-HT bioavailability. Information about the occurrence of the reward-predictive stimulus was maximized during reward consumption, around 3 to 4 s after the presentation of the cue, and it was higher under chronic fluoxetine treatment. However, the encoded information was less robust to noise corruption when compared to control conditions.

Chronic Fluoxetine Enhances Extinction Therapy for PTSD by Evaluating Brain Glucose Metabolism in Rats: An [18F]FDG PET Study

Recent studies suggest that selective serotonin reuptake inhibitors (SSRIs) and exposure therapies have been used to reduced footshock-induced posttraumatic stress disorder (PTSD) symptoms. However, the therapeutic effect of the combination of SSRIs treatment with exposure therapy remains a matter of debate.This study aimed to evaluate these therapeutic effect through the behavioral and the neuroimaging changes by positron emission tomography (PET) in model rats. We adopted a Pavlovian fear conditioning paradigm to establish model rats, and serial [18F]fluorodeoxyglucose (FDG) PET imaging was performed to investigate the neurofunctional changes before and after the therapies (chronic fluoxetine and/or extinction). Behavioral tests indicated combined therapy group performanced lower freezing (%) than the two other groups.Compared with the baseline, fear conditioning increased 18F-FDG uptake in the right amygdala but decreased uptake in the left primary somatosensory cortex. After extinction retrieval, there was increased [18F]FDG uptake in the left striatum but decreased uptake in the anterior cingulate cortex in the extinction group. Fluoxetine increased [18F]FDG uptake in the left hippocampus but decreased uptake in the cuneiform nucleus. The combined therapy increased [18F]FDG uptake in the left hippocampus, left striatum, right insular cortex but reduced uptake in the cerebellar lobule. c-Fos expression was significantly higher in the hippocampal dentate gyrus and anterior cingulate cortex in the fluoxetine and combined groups than that in the extinction group, with no significant difference between the two groups. Taken together, chronic fluoxetine enhanced the effects of extinction training in a rat model of PTSD. In vivo PET imaging may provide a promising approach for evaluation chronic fluoxetine treatment of PTSD.

Neuroinflammation-Induced Parvalbumin Interneuron and Oscillation Deficits Might Contribute to Neurobehavioral Abnormities in a Two-Hit Model of Depression

Background: Depression is a common neuropsychiatric disorder that causes profound disability worldwide, yet the underlying mechanism remains unclear. Thus, the present study aimed to evaluate the effects of a two-hit model of depression on glial activation, parvalbumin (PV) interneuron, oscillation activity, and behavior alternations, and whether chronic fluoxetine treatment can reverse these abnormalities. Methods: Male mice were submitted to lipopolysaccharide (LPS) injection, followed by a modified chronic unpredictable stress (CUS) protocol. Results: In our study, we showed that mice exposed to LPS and CUS exhibited reduced body weight, anhedonic-like behavior as well as cognitive and anxiety symptoms. These behavioral alternations were related to enhanced neuroinflammation, as reflected by significantly increased IL-1β and IL-6 levels and microglia activation in the prefrontal cortex (PFC). In addition, mice exposed to LPS and CUS displayed significantly decreased PV expression and disturbance of theta and gamma oscillations in the PFC. However, chronic fluoxetine treatment reversed most of these abnormalities. Conclusion: Our study of this two-hit model of depression is clinically relevant and suggests the combination of different etiological and pathophysiological components of depression may provide with a more translational value.

Basolateral Amygdala but Not Medial Prefrontal Cortex Contributes to Chronic Fluoxetine Treatments for PTSD Symptoms in Mice

Do chronic fluoxetine treatments reduced footshock-induced posttraumatic stress disorder (PTSD) symptoms, including fear and comorbid depression, in the situational reminder phase? Moreover, are the subareas of the medial prefrontal cortex (mPFC), including the cingulate cortex 1 (Cg1), prelimbic cortex (PrL), infralimbic cortex (IL), and basolateral amygdala (BLA), involved in the fluoxetine amelioration of PTSD symptoms? These two crucial issues were addressed in the present study. All mice were injected with chronic fluoxetine or normal saline treatments for the adaptation (14 days), footshock fear conditioning (1 day), and situational reminder (3 days) phases. After adaptation, the mice were subjected to footshock (2 mA, 10 seconds) or nonfootshock and stayed 2 min in a footshock box for 2 min for fear conditioning. Later, they were placed in the footshock box for 2 min in the situational reminder phase. In the final session of the situational reminder phase, a forced swimming test (FST) and immunohistochemical staining were conducted. The results indicated that footshock induced fear and comorbid depression. Meanwhile, chronic fluoxetine treatments reduced fear and depression behaviors. The Cg1, PrL, IL, and BLA were seemingly to increase c-Fos expression after footshock-induced PTSD symptoms in the situational reminder phase. The fluoxetine treatments reduced only the BLA’s c-Fos expression. The findings suggest that BLA contributes to the fluoxetine amelioration of PTSD symptoms; however, the mPFC, including the Cg1, PrL, and IL, did not mediate PTSD symptoms’ amelioration stemming from fluoxetine. The present data might help us to further understand the neural mechanism of fluoxetine treatments in PTSD symptoms.