ABSTRACT
Major depressive disorder is highly prevalent worldwide and has been affecting an increasing number of people each year. Current first line antidepressants show merely 37% remission, and physicians are forced to use a trial-and-error approach when choosing a single antidepressant out of dozens of available medications. We sought to identify a method of testing that would provide patient-specific information on whether a patient will respond to a medication using in vitro modeling. Patient-derived lymphoblastoid cell lines from the Sequenced Treatment Alternatives to Relieve Depression study were used to rapidly generate cortical neurons and screen them for bupropion effects, for which the donor patients showed remission or non-remission. We provide evidence for biomarkers specific for bupropion response, including synaptic connectivity and morphology changes as well as specific gene expression alterations. These biomarkers support the concept of personalized antidepressant treatment based on in vitro platforms and could be utilized as predictors to patient response in the clinic.
Subject(s)
Depressive Disorder, Major , Antidepressive Agents/therapeutic use , Biomarkers , Depression , Depressive Disorder, Major/drug therapy , Humans , Neurons , Treatment OutcomeABSTRACT
We investigated the efficacy of optogenetic inhibition at presynaptic terminals using halorhodopsin, archaerhodopsin and chloride-conducting channelrhodopsins. Precisely timed activation of both archaerhodopsin and halorhodpsin at presynaptic terminals attenuated evoked release. However, sustained archaerhodopsin activation was paradoxically associated with increased spontaneous release. Activation of chloride-conducting channelrhodopsins triggered neurotransmitter release upon light onset. Thus, the biophysical properties of presynaptic terminals dictate unique boundary conditions for optogenetic manipulation.
Subject(s)
Biophysical Phenomena/physiology , Neural Inhibition/physiology , Optogenetics/methods , Presynaptic Terminals/metabolism , Synaptic Transmission/physiology , Animals , Cells, Cultured , Female , HEK293 Cells , Halorhodopsins/metabolism , Hippocampus/metabolism , Humans , Male , Mice , Mice, Inbred C57BL , Organ Culture Techniques , Rats , Rats, Sprague-DawleyABSTRACT
Protein kinase M zeta (PKMζ), an atypical isoform of protein kinase C (PKC), has been implicated in long-term maintenance of neuronal plasticity and memory. However, the cellular machinery involved in these functions has yet to be elucidated. Here, we investigated the effects of PKMζ overexpression on the morphology and function of cortical neurons in primary cultures. Transfection with a plasmid construct expressing the PKMζ gene modified the distribution of spine morphologies and reduced spine length, while leaving total spine density and dendritic branching unchanged. A significant increase in magnitude but not frequency of miniature excitatory post synaptic currents was detected in the PKMζ overexpressing cells. These results suggest that PKMζ is involved in regulation of dendritic spine structure and function, which may underlie its role in long-term synaptic and behavioral plasticity.
Subject(s)
Cerebral Cortex/enzymology , Cerebral Cortex/ultrastructure , Dendritic Spines/ultrastructure , Neurons/enzymology , Neurons/ultrastructure , Protein Kinase C/biosynthesis , Animals , Cell Size , Cell Survival , Electrophysiological Phenomena , Excitatory Postsynaptic Potentials/genetics , Excitatory Postsynaptic Potentials/physiology , Gene Expression Regulation, Enzymologic/physiology , Immunohistochemistry , Neuronal Plasticity/physiology , Plasmids/genetics , Protein Kinase C/physiology , Rats , Synapses/physiology , TransfectionABSTRACT
Memories are more easily disrupted than improved. Many agents can impair memories during encoding and consolidation. In contrast, the armamentarium of potential memory enhancers is so far rather modest. Moreover, the effect of the latter appears to be limited to enhancing new memories during encoding and the initial period of cellular consolidation, which can last from a few minutes to hours after learning. Here, we report that overexpression in the rat neocortex of the protein kinase C isozyme protein kinase Mζ (PKMζ) enhances long-term memory, whereas a dominant negative PKMζ disrupts memory, even long after memory has been formed.
Subject(s)
Memory, Long-Term , Neocortex/metabolism , Protein Kinase C/genetics , Protein Kinase C/metabolism , Animals , Conditioning, Psychological , Gene Expression , Gene Transfer Techniques , Genetic Vectors , Isoenzymes/genetics , Isoenzymes/metabolism , Lentivirus/genetics , Male , Mutant Proteins/metabolism , Mutation , Neurons/metabolism , Rats , Rats, WistarABSTRACT
The purpose was to describe blood flow waveform of pelvic organs obtained by Doppler according to their unique characteristics. A prospective study was designed and 79 premenopausal and postmenopausal women were screened. Transvaginal ultrasonography combined with color Doppler was performed. Arterial blood flow of the uterus, fallopian tubes and both ovarian center and periphery were assessed, by a unique computerized program exclusively developed for this research (MATLAB language). Waveform characterization was performed by calculating alpha and beta angles, representing upward curve of each waveform and angles of refraction gamma and delta. alpha to delta angles were found significantly different for each of the pelvic organs. Significant differences in the characteristics of Doppler waveforms were also observed between pre and postmenopausal women. Luteal and follicular phase blood flow waveforms were similar. These findings contribute to our ability to classify the origin of blood vessel by processing Doppler waveforms by a computerized method.
