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McLean Hospital

7 ARTICLES PUBLISHED IN JoVE

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Biology

Obtaining High Quality RNA from Single Cell Populations in Human Postmortem Brain Tissue
Charmaine Y. Pietersen 1, Maribel P. Lim 1, Tsung-Ung W. Woo 1,2,3
1Department of Structural and Molecular Neuroscience, McLean Hospital, 2Department of Psychiatry, Harvard Medical School, 3Department of Psychiatry, Beth Israel Deaconess Medical Center

We describe a process using laser-capture microdissection to isolate and extract RNA from a homogeneous cell population, pyramidal neurons, in layer III of the superior temporal gyrus in postmortem human brains. We subsequently linearly amplify (T7-based) mRNA, and hybridize the sample to the Affymetrix human X3P microarray.

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Neuroscience

Methods for Studying the Mechanisms of Action of Antipsychotic Drugs in Caenorhabditis elegans
Limin Hao 1,2, Edgar A. Buttner 1,2
1Department of Psychiatry, Harvard Medical School, 2Mailman Research Center, McLean Hospital

Approaches for testing the effects of antipsychotic drugs (APDs) in Caenorhabditis elegans are demonstrated. Assays are described for testing drug effects on development and viability and on pharyngeal pumping rate. These methods are also applicable for pharmacogenetic experiments with drug classes other than APDs.

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Neuroscience

Isolation and Culture of Endothelial Cells from the Embryonic Forebrain
Peeyush Kumar T. 1,2, Anju Vasudevan 1,2
1Department of Psychiatry, Harvard Medical School, 2Angiogenesis and Brain Development Laboratory, Division of Basic Neuroscience, McLean Hospital

This video demonstrates an easy and reliable strategy for preparation of pure cultures of endothelial cells from the embryonic forebrain within 10-12 days and will be useful for research focused on many aspects of cerebral angiogenesis.

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Biology

Production of Nurr-1 Specific Polyclonal Antibodies Free of Cross-reactivity Against Its Close Homologs, Nor1 and Nur77
Pierre Leblanc 1, Minho Moon 1,2, Woori Kim 1, Inhye Jeong 1, Chun-Hyung Kim 1, Kwang-Soo Kim 1
1Molecular Neurobiology Laboratory, Psychiatry, McLean Hospital, 2Department of Biochemistry, College of Medicine, Konyang University

Here, we selectively target antibodies against a specific member of a highly conserved family of proteins by immunizing animals with their most divergent regions followed by removing cross reactive antibodies by pre-adsorption.

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Neuroscience

Migration, Chemo-Attraction, and Co-Culture Assays for Human Stem Cell-Derived Endothelial Cells and GABAergic Neurons
Debkanya Datta 1,2, Anju Vasudevan 1,2
1Department of Psychiatry, Harvard Medical School, 2Angiogenesis and Brain Development Laboratory, Division of Basic Neuroscience, McLean Hospital

We present three simple in vitro assays-the long-distance migration assay, the co-culture migration assay, and chemo-attraction assay-that collectively evaluate the functions of human stem cell derived periventricular endothelial cells and their interaction with GABAergic interneurons.

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Neuroscience

Derivation, Expansion, Cryopreservation and Characterization of Brain Microvascular Endothelial Cells from Human Induced Pluripotent Stem Cells
Sovannarath Pong 1,2,3, Paulo Lizano 1,2,3,4, Rakesh Karmacharya 1,3,4,5
1Center for Genomic Medicine, Massachusetts General Hospital, 2Department of Psychiatry, Beth Israel Deaconess Medical Center, 3Chemical Biology and Therapeutic Science Program, Broad Institute of MIT and Harvard, 4Department of Psychiatry, Harvard Medical School, 5Schizophrenia and Bipolar Disorder Program, McLean Hospital

This protocol details an adapted method to derive, expand, and cryopreserve brain microvascular endothelial cells obtained by differentiating human induced pluripotent stem cells, and to study blood brain barrier properties in an ex vivo model.

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Neuroscience

Co-Culturing Microglia and Cortical Neurons Differentiated from Human Induced Pluripotent Stem Cells
Kara Lopez-Lengowski 1,2, Annie Kathuria 1,2,3, Kaia Gerlovin 1,2, Rakesh Karmacharya 1,2,3,4,5,6,7
1Center for Genomic Medicine, Massachusetts General Hospital, 2Chemical Biology Program, Broad Institute of MIT & Harvard, 3Department of Psychiatry, Harvard Medical School, 4Schizophrenia and Bipolar Disorder Program, McLean Hospital, 5Program in Neuroscience, Harvard University, 6Program in Chemical Biology, Harvard University, 7Harvard Stem Cell Institute

This protocol describes a methodology to differentiate microglia from human iPSCs and maintain them in co-culture with iPSC-derived cortical neurons in order to study mechanistic underpinnings of neuroimmune interactions using human neurons and microglia.

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