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1.
Cell ; 174(2): 465-480.e22, 2018 07 12.
Artículo en Inglés | MEDLINE | ID: mdl-30007418

RESUMEN

Modern genetic approaches are powerful in providing access to diverse cell types in the brain and facilitating the study of their function. Here, we report a large set of driver and reporter transgenic mouse lines, including 23 new driver lines targeting a variety of cortical and subcortical cell populations and 26 new reporter lines expressing an array of molecular tools. In particular, we describe the TIGRE2.0 transgenic platform and introduce Cre-dependent reporter lines that enable optical physiology, optogenetics, and sparse labeling of genetically defined cell populations. TIGRE2.0 reporters broke the barrier in transgene expression level of single-copy targeted-insertion transgenesis in a wide range of neuronal types, along with additional advantage of a simplified breeding strategy compared to our first-generation TIGRE lines. These novel transgenic lines greatly expand the repertoire of high-precision genetic tools available to effectively identify, monitor, and manipulate distinct cell types in the mouse brain.


Asunto(s)
Encéfalo/metabolismo , Técnicas de Inactivación de Genes/métodos , Genes Reporteros , Animales , Encéfalo/citología , Calcio/metabolismo , Línea Celular , Hibridación Fluorescente in Situ , Luz , Ratones , Ratones Transgénicos , Microscopía Fluorescente , Neuronas/metabolismo , Optogenética , ARN no Traducido/genética , Transgenes/genética
2.
Nature ; 563(7729): 72-78, 2018 11.
Artículo en Inglés | MEDLINE | ID: mdl-30382198

RESUMEN

The neocortex contains a multitude of cell types that are segregated into layers and functionally distinct areas. To investigate the diversity of cell types across the mouse neocortex, here we analysed 23,822 cells from two areas at distant poles of the mouse neocortex: the primary visual cortex and the anterior lateral motor cortex. We define 133 transcriptomic cell types by deep, single-cell RNA sequencing. Nearly all types of GABA (γ-aminobutyric acid)-containing neurons are shared across both areas, whereas most types of glutamatergic neurons were found in one of the two areas. By combining single-cell RNA sequencing and retrograde labelling, we match transcriptomic types of glutamatergic neurons to their long-range projection specificity. Our study establishes a combined transcriptomic and projectional taxonomy of cortical cell types from functionally distinct areas of the adult mouse cortex.


Asunto(s)
Perfilación de la Expresión Génica , Neocórtex/citología , Neocórtex/metabolismo , Animales , Biomarcadores/análisis , Femenino , Neuronas GABAérgicas/metabolismo , Ácido Glutámico/metabolismo , Masculino , Ratones , Corteza Motora/anatomía & histología , Corteza Motora/citología , Corteza Motora/metabolismo , Neocórtex/anatomía & histología , Especificidad de Órganos , Análisis de Secuencia de ARN , Análisis de la Célula Individual , Corteza Visual/anatomía & histología , Corteza Visual/citología , Corteza Visual/metabolismo
3.
J Comp Neurol ; 530(1): 6-503, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34525221

RESUMEN

Increasing interest in studies of prenatal human brain development, particularly using new single-cell genomics and anatomical technologies to create cell atlases, creates a strong need for accurate and detailed anatomical reference atlases. In this study, we present two cellular-resolution digital anatomical atlases for prenatal human brain at postconceptional weeks (PCW) 15 and 21. Both atlases were annotated on sequential Nissl-stained sections covering brain-wide structures on the basis of combined analysis of cytoarchitecture, acetylcholinesterase staining, and an extensive marker gene expression dataset. This high information content dataset allowed reliable and accurate demarcation of developing cortical and subcortical structures and their subdivisions. Furthermore, using the anatomical atlases as a guide, spatial expression of 37 and 5 genes from the brains, respectively, at PCW 15 and 21 was annotated, illustrating reliable marker genes for many developing brain structures. Finally, the present study uncovered several novel developmental features, such as the lack of an outer subventricular zone in the hippocampal formation and entorhinal cortex, and the apparent extension of both cortical (excitatory) and subcortical (inhibitory) progenitors into the prenatal olfactory bulb. These comprehensive atlases provide useful tools for visualization, segmentation, targeting, imaging, and interpretation of brain structures of prenatal human brain, and for guiding and interpreting the next generation of cell census and connectome studies.


Asunto(s)
Atlas como Asunto , Encéfalo/crecimiento & desarrollo , Corteza Entorrinal/crecimiento & desarrollo , Hipocampo/crecimiento & desarrollo , Animales , Femenino , Humanos , Embarazo
4.
eNeuro ; 4(5)2017.
Artículo en Inglés | MEDLINE | ID: mdl-28932809

RESUMEN

Transgenic mouse lines are invaluable tools for neuroscience but, as with any technique, care must be taken to ensure that the tool itself does not unduly affect the system under study. Here we report aberrant electrical activity, similar to interictal spikes, and accompanying fluorescence events in some genotypes of transgenic mice expressing GCaMP6 genetically encoded calcium sensors. These epileptiform events have been observed particularly, but not exclusively, in mice with Emx1-Cre and Ai93 transgenes, of either sex, across multiple laboratories. The events occur at >0.1 Hz, are very large in amplitude (>1.0 mV local field potentials, >10% df/f widefield imaging signals), and typically cover large regions of cortex. Many properties of neuronal responses and behavior seem normal despite these events, although rare subjects exhibit overt generalized seizures. The underlying mechanisms of this phenomenon remain unclear, but we speculate about possible causes on the basis of diverse observations. We encourage researchers to be aware of these activity patterns while interpreting neuronal recordings from affected mouse lines and when considering which lines to study.


Asunto(s)
Calcio/metabolismo , Corteza Cerebral/fisiopatología , Epilepsia , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Neuronas/fisiología , Animales , Corteza Cerebral/diagnóstico por imagen , Corteza Cerebral/metabolismo , Modelos Animales de Enfermedad , Doxiciclina/farmacología , Epilepsia/genética , Epilepsia/patología , Epilepsia/fisiopatología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Integrasas , Ratones , Ratones Transgénicos
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