sCRACM: ChR2 circuit mapping

As has become a hallmark of the Svoboda lab, this new paper in Nature (advance online publication) combines several cutting edge technologies (rAAV-delivered ChR2, most prominently, and 2-photon 1-photon laser stimulation) to do some interesting synaptic physiology.

The subcellular organization of neocortical excitatory connections : Article : Nature.

They used ChR2 (with TTX and 4-AP to block action potentials) to find where on the dendritic tree particular inputs synapsed onto L3 and L5 cells and to measure the strength of those inputs. ChR2 depolarizes the input axon locally (60um spot diameter) at points of (potential) axodendritic contact. If you’ve heard the term “potential synapse” before, then think of this technique as a way of checking potential synapses and seeing if there really is an actual synapse there.

The technique allowed them to map on a L3 barrel cortex pyramidal cell where different thalamic inputs (VPm, POm) and cortical inputs (M1, barrel L2/3, barrel L4):

screenshot001

sCRACM stands for subcellular ChR2-assisted circuit mapping.

Plant neuroscience

Plants Found to Show Preferences for Their Relatives – NYTimes.com

Two amazing things here:

  1. Plants missing photosynthetic enzymes of their own that migrate directionally toward “victim” plants. This behavior has an uncanny resemblance to axon guidance. Make sure to view the time-lapse video in the NYT article. Here’s an image from the PSU website:

  2. Plants capable of identifying kin and “being nice” to kin while going into a competitive mode of root growth with non-kin. Amazing.

It refreshing to see this kind of interesting behavior without any neurons involved. It makes me think (realize) that the idea of a neuron or a neural system has many components and there might not be any good reason to assume that a single cell must have all of those properties or none of them. Something like a neuron-like cell that’s not a neuron in the classical sense. Anyone know of other examples?

Split GFP reconstituted: A dynamic synapse label

This new technique from Cori Bargmann’s lab is one of the neatest that I’ve seen in a while. The authors split GFP into two pieces, expressing one piece presynaptically and the other postsynaptically. This creates functional (ie. fluorescing) GFP only at sites of synaptic contact where the protein can reconstitute. They call the technique GFP Reconstitution Across Synaptic Partners (GRASP). Check out an example labeling here:
GRASP labeling figure
The neurons are expressing mCherry in the cytoplasm but GFP is expressed only at the site of synaptic contacts where the split GFP peptides can be reconstituted into a complete GFP fluorophore.

Real-time STED to visualize vesicle dynamics

Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement

Just the optical engineering alone here deserves mention: 28 frames per second at 62nm resolution (well below the diffraction limit of 260nm for light of the wavelength used)! STED (or stimulated emission depletion, developed in Stefan Hell’s group) is ideal for visualizing synaptic vesicles, whose small size (~50nm) has typically confined them to the domain of electron microscopists. The ability to get high-speed STED allowed the researchers to track individual vesicles and their path dynamics. They conclude that vesicle movement has both motor-driven and diffusive components (ie. a biased random walk). I’m sure with more time and more analysis there will be a lot of interesting applications for this kind of real-time vesicle tracking. Perhaps in the near future we will have single vesicle “minis” monitored at multiple sites through microscopy instead of just one or two sites electrophysiologically…

Here’s the resolution difference between STED and confocal for a single vesicle:
Sted vs. confocal vesicle picture

And, for those of you with ~$1.25M lying around, you can now purchase a STED setup directly from Leica!

Postdoctoral positions at Janelia Farm

Postdoctoral/research scientist positions are available in the inter-disciplinary group of Dmitri Chklovskii at the new Janelia Farm Research Campus of the Howard Hughes Medical Institute located in the suburbs of Washington, D.C. Candidates are expected to have a PhD in neuroscience, physics, computer science or electrical engineering. Most of the work is theoretical or computational and is done in collaboration with several experimental laboratories. Successful applicants will work on projects centered on neuronal circuits such as high-throughput reconstruction of wiring diagrams as well as combining structural and physiological data to infer circuit function. Salary will be commensurate with qualifications. For more information about research directions in the group please see: http://www.hhmi.org/research/groupleaders/chklovskii.html
Interested applicants should send their CV and a statement of research interests to mitya (at) janelia.hhmi.org, and arrange for three recommendation letters to be emailed to me.

Amputee Controls And Feels Bionic Arm as Her Own

(UPDATE 03-05-2007 – Upon closer inspection, it is clear that while the surgery has enabled the woman to have sensation in the nerves of her missing hand when the surface of her chest is touched, the arm she is fitted with at the time of publication did not relay sensory signals from the arm back to her chest. As soon as she is fitted with an arm that has the appropriate sensors, however, she will not have to undergo further surgery to have this kind of direct feedback. Thanks to astute readers for pointing this out.)

The Guardian reports on an article published today in the Lancet about a successful surgical procedure giving an amputee a bionic arm that both responds to motor commands from her remaining motor nerves to control it and provides sensory feedback to sensory nerves when it is touched. If there was any doubt left, the worlds of neural prosthetics and brain-machine interfaces have officially collided.

The Lancet article is accompanied by two movies of the woman using the arm that you should really check out.

Given the recent progress in the decoding of motor signals from the brain and older progress on sensory feedback from neural prosthetics, this was to be expected. Nonetheless, watching this woman use her arm brings the message home in a visceral way. The spooky thesis of MIT CSAIL’s Rodney Brooks that “we will become a merger between flesh and machines” is one step closer today.

Continue reading

Spontaneous Rewiring seen in 4 hrs.

It seems Markram is again back to getting some interesting results. Recently a new discovery from the Brain Mind Institute of the EPFL shows that the brain adapts to new experience by unleashing a burst of new neuronal connections, and only the fittest survive. The research further shows that this process of creation, testing, and reconfiguring of brain circuits takes place on a scale of just hours, suggesting that the brain is evolving considerably even during the course of a single day.

The paper can be found Here.

Presynaptic somatic membrane potential can influence EPSPs

Modulation of intracortical synaptic potentials by presynaptic somatic membrane potential : Nature

Very interesting work. Modulation of the somatic potential seems to influence the EPSP, as measured by paired patch recordings of two layer 5 cells in cortical slice. Somatic depolarization from resting potential to near threshold results in an increase in evoked EPSPs.

In synaptic physiology, we often make a point of distinguishing intrinsic changes (eg. membrane potential) from synaptic conductance changes. Now it looks like the line between those might be a bit blurry!

Here’s a N&V by Eve Marder too.

Synaptic tuning : Nature Reviews Neuroscience

Synaptic tuning : Nature Reviews Neuroscience

For those interested in neuromodulators:

Treatment of striatal neurons with a D1 receptor agonist led to an increase in the dendritic staining intensity of NMDA receptor NR2B subunits. There was also an increase in the association of NR2B subunits with PSD-95 — a scaffold protein required for the assembly of NMDA receptors — and in the surface localization of NR2B-containing receptors.

Original article in J. Neurosci. from Dunah and colleagues. An excerpt from the original aricle of a neat application of FRET continues after the jump.
Continue reading