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Using AAV for neuronal tracing

Posted by Klaus Wanisch on Aug 9, 2018 9:04:52 AM

Background on neuronal tracing

A key aspect to understanding the brain’s function is knowing its architecture, in particular the connections between different brain regions. For example, communication between the hippocampus and the prefrontal cortex brain regions is involved in the formation of episodic memory, a special type of memory which includes autobiographical events (see Jin & Maren, 2015). Directional flow of information between different parts of the brain is mediated via individual neurons. Neurons are composed of a cell body, with dendrites receiving incoming information, and a projecting axon sending information onwards to other neuronal cells. Synapses at the terminals of axons form connections to dendrites of proximal neuronal cells. In the specific example of episodic memory, a subset of hippocampal neurons projects axons directly to the prefrontal cortex, but also indirectly via synapses to neurons in other brain regions. Further, the connections between regions are often reciprocal, forming a neuronal loop which is activated and strengthened during memory formation and memory retrieval.

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Topics: Viral Vectors, Cell Tracing, AAV

Rabies and Neuronal Tracing

Posted by Leila Haery on May 29, 2018 9:51:06 AM

Why study neural connectivity?

One of the early lessons many of us learned in biology is that the body’s architecture and plumbing are important. We started with learning the head is connected to the neck. Shortly after, we learned about organs and the jobs they perform. This became foundational later on when we studied biological processes, like how our stem cells are housed in specific locations and give rise to progenitors during growth and development or that blood flows through the heart and lungs and oxygenates the body. However, in neuroscience, this architecture is frequently still an open question. The connections between neurons are what define how the brain operates, and thus, are a major part of the answer to many biological questions about the brain. To address this, molecular tools to map neuronal connectivity are widely used in neuroscience. In this post, I’ll describe how rabies virus (RABV) can be used in the brain to visualize how neurons are connected.

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Topics: Viral Vectors, Cell Tracing

Retrograde AAV: Making the Journey from Axon to Nucleus

Posted by Leila Haery on May 16, 2017 10:30:00 AM

The concept that the brain has a structure is not obvious. While it’s been a long time since Aristotle argued the heart was the thought center of the body, it wasn’t until the 1700s that scientists hypothesized and began to gather evidence that the brain has distinct regions with specialized functions. Phineas Gage, the man whose personality changed drastically after an accident where an iron spike was driven through his head, is a famous early example of the link between brain regions and behavior.  Also around that time, French scientists Marc Dax and Paul Broca independently discovered the speech production center of the brain when autopsies of speech-impaired patients revealed lesions in a particular brain region, later named the Broca’s area. In this post I’ll describe a new virus with retrograde function and how it’s enabling scientists to access neurons in a powerful way. Keep reading to find out what retrograde function is and how it gives us better access and ultimately a better understanding of the brain.

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Topics: Viral Vectors, Cell Tracing, AAV, Neuroscience

New Tool for Lineage Tracing: The ClonTracer Library

Posted by Tyler Ford on Sep 22, 2015 10:30:00 AM

This article is based on an interview with Novartis researcher, Carrie Bhang.

The ClonTracer Library, deposited by Carrie Bhang, a research investigator in the In Vivo Pharmacology group at Novartis Oncology, is an exciting new tool that allows researchers to individually label millions of mammalian cells through lentiviral infection and to monitor their abundance and clonal dynamics over time using next generation sequencing (NGS). The library was developed when Carrie was a post-doc in Frank Stegmeier’s lab in Novartis Oncology. 

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Topics: Viral Vectors, Cancer, Cell Tracing, Retroviral and Lentiviral Vectors

Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

Posted by Mary Gearing on Apr 24, 2015 10:39:00 AM

CRISPR-Cas9 genome editing may be the hot new way to manipulate gene expression, but other gene manipulation systems remain valuable to biology. Cre-lox recombination, discovered in the 1980s, is one of the most important ways to spatially and temporally control gene expression, especially in in vivo models, and new Cre-lox based technologies are still being developed today. In this post, I will highlight the evolution of the  Brainbow multicolor labeling system - a perfect example of the continued utility of Cre-lox. Check out our previous blog post, Plasmids 101: Cre-lox, if you need a quick primer on how Cre-lox recombination works.

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Topics: Fluorescent Proteins, Fluorescent Imaging, Cell Tracing, Neuroscience

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