New Lab

We, Katz lab, and the Neuroscience Institute have moved to the new building near Georgia State MARTA Station.
We are on the 8th floor.


Cool-looking building. Tornado-proof? Probably not.

My rig room has a window. I can see the horizon while poking cells!


Our aquarium is now 30 sec away from the lab. This is great.
I did lots of plumbing work to install new filtering system and protein skimmers. This is a hard work. I have had 3 leakage from various PVC joints I made. One of them was due to a crack that was developed during weekend. By Monday morning, half of the tank water leaked out.

One of the chillers failed because the compressor overload protector had fallen off and disconnected the power. I opened it up the chiller and rewired the circuit to skip this failed protector. Now it is running fine.

I was also in panic when I found the other chiller heated up in the morning. The radiator fan was not running. The water was like hot tub. I opened the chiller and found that the fan blade was stuck by touching the frame. I mended it back and now its working fine like new.

Setting up the aquarium is like buying a new Harley Davidson. You will have trouble after trouble until the machinery gets settled.
I would definitely buy Honda.

Backfill

I did backfills on two Melibe brains. Both appeared very nicely.

My success rate on backfilling is pretty high, almost 100%.
It is simple. All you have to do is to let dye solution touch only the cut end of the nerve you want to fill. And make sure the prep is in healthy condition until fixation. Poor results are mostly due to failure in these two things. It is not because of materials or protocols. It is how you manipulate them.

Plasticity during stroke recovery: from synapse to behavior

Timothy H. Murphy and Dale Corbett
Nature Reviews in Neuroscience 10: 861-872 (2009)

Role of ipsilateral pathways
Although canonical view of sensory and motor processing is that body parts are controlled by neurons in the cerebral hemisphere on the opposite side of the body, ipsilateral pathways are also present.

(although the redundancy of an unaffected cortex and the potential of ipsilateral pathways seem advantageous, the issues of lateralization and function are potentially complex and reflect both the degree of injury and the extent of recovery)

Diffuse connectivity
- surprisingly widespread intracortical connectivity between related regions of the cortex, ...

Location is everyting in cortical physiology
- much like the demand for a vacant lot in Manhattan, ...
- there is intense competetion for available cortical map territory
- How the remapping of lost function is initiated, and how seemingly stroke-compromised circuits in the peri-infarct cortex can compete and win in what is thought to be an activity-dependent process, is unclear.

---

Synaptic learning rules in recovery
synapse-based learning rules could help to create compensatory circuits after stroke (Fig. 3).

Homeostatic plasticity and Hebbian plasticity mechanisms
There are no direct evidence for their contribution in recovery.

Insights into a molecular switch that gates sensory neuron synapses during habituation in Aplysia

Tony D. Gover, Thomas W. Abrams

I think this paper spends so much space and time only to deny depletion hypothesis.
The effects of calpain and Arf sound very interesting and promissing.


1. Habituation of reflex responses in vertebrates and invertebrates: a simple form of neural plasticity
In addition to habituation, the repeated stimulus may lead to a strengthening of the response, known as sensitization.
Their observations led them to propose the ‘‘dual-process theory” in which the behavioral change actually reflects the
contribution of these two independent processes.

Whereas short-term depression at these synapses appears to be mediated entirely by presynaptic changes (Castellucci & Kandel, 1974; Manseau, Fan, Hueftlein, Sossin, & Castellucci, 2001), long-term depression that is initiated by extensive stimulation or blocks of stimuli delivered over hours appears to depend upon signals initiated by postsynaptic glutamate receptors, including Ca2+ influx through NMDA receptors (Lin & Glanzman, 1996; Mata, Chen, Cai, & Glanzman, 2008) (see also Ezzeddine & Glanzman, 2003).

2. Mechanisms of synaptic depression in other systems

2.1. Depletion of the readily releasable pool of vesicles as a mechanism of depression
Historically, the earliest proposed mechanism for synaptic depression was depletion of the neurotransmitter that is available for release. Vesicles at synaptic release sites that are available for immediate release are considered to constitute a readily releasable pool (Betz, 1970; Gingrich & Byrne, 1985; Stevens & Tsujimoto, 1995); these vesicles may already be docked at exocytosis sites.

2.2. Use-dependent inactivation of presynaptic Ca2+ channels

2.3. Desensitization of postsynaptic receptors

2.4. Autoreceptors and retrograde signaling
At parallel fiber-to-Purkinje cell synapses in cerebellum, high frequency presynaptic activity causes suppression of transmitter release, which is mediated by release of endocannabinoids from the postsynaptic Purkinje cell; the endocannabinoids act on receptors on presynaptic terminals, resulting in reduced Ca²⁺ influx (Brown, Brenowitz, & Regehr, 2003).

3. Proposed mechanisms for synaptic depression at the sensory neuron-to-motor neuron synapse in Aplysia
3.1. Short-term homosynaptic depression at sensory neuron-to-motor neuron synapses involves a presynaptic mechanism, rather than alteration in postsynaptic responses

3.2. Ca2+ current inactivation
With repeated stimulation of the sensory neuron, the action potential duration decreased, suggesting that Ca2+ influx decreased with each successive action potential. Klein et al. (1980) also measured the Ca²⁺ current in
the soma of the presynaptic sensory neuron under voltage clamp.
With repeated depolarizing voltage steps, they observed that the Ca²⁺ current decreased in parallel with the decline in the amplitude of the EPSP recorded in the motor neuron.
Gingrich and Byrne (1985) also reached the conclusion that Ca²⁺ current inactivation alone during the normal duration ac-tion potential does not account for depression at this synapse.

Armitage and Siegelbaum (1998) used fluorescent Ca²⁺ indicators to directly measure Ca²⁺ influx at presynaptic varicosities of sensory neurons through the dihydropyridine-insensitive Ca²⁺ channels. No change in Ca²⁺ influx through these channels that initiate release was observed during synaptic depression.

3.3. Depletion of the readily releasable pool of vesicles as the primary mechanism for synaptic depression during habituation
Thus, depletion of the readily releasable pool of vesicles can contribute to depression, but probably only with relatively prolonged presynaptic activity.
Recent evidence implicated the Ca²⁺-activated protease calpain both in modulation of depression at the sensory neuron-to-motor neuron synapse and in serotonin-induced facilitation after depression (Khoutorsky & Spira, 2005).
This intriguing observation has been interpreted to indicate that this protease acts to untether vesicles in the reserve
pool, and thereby initiates mobilization of vesicles to the readily releasable pool. Although this is a plausible model, another possible function for calpain is the proteolytic cleavage of protein kinase C (PKC), converting the enzyme to the persistently active PKM form (Sutton, Bagnall, Sharma, Shobe, & Carew, 2004). PKC is known to participate in facilitation of depressed sensory neuron-to-motor neuron synapses by serotonin (Ghirardi et al., 1992; Manseau
et al., 2001), and also acts to prevent the development of depression during bursts of activity (see below). Thus, calpain cleavage of PKC could reduce synaptic depression.

3.4. Switching of release sites to a silent state as a mechanism of synaptic depression

Gover et al. (2002) considered five mechanisms of synaptic depression:
(1) vesicle depletion, which by default must be dependent on vesicular release (Fig. 3);
(2) reduction in the probability of release of individual releasable vesicles after a release event;
(3) reduction in the probability of release of individual releasable vesicles after an action potential, but independent of a release event;
(4) silencing of release sites after a release event, and
(5) silencing of release sites, after an action potential, but independent of a release event (Fig. 4).

These data suggest that the probability of release does not change gradually; rather individual release sites are simply silenced, or switched off, during development of depression.
Furthermore, Royer et al. (2000) argued that given their data, if p were nonuniform across release sites, then homosynaptic depression due to silencing of release sites would be independent of release [which is consistent with the conclusions of Gover et al. (2002)].

3.5. Other evidence suggesting depletion of vesicles is not the major mechanism for synaptic depression at Aplysia sensory neuron synapses


4. Evidence from other systems suggesting that vesicle depletion is not the primary mechanism responsible for synaptic depression
Rather depression with relatively few stimuli or with low frequency stimulation cannot be explained primarily by depletion.

5. Ca²⁺ influx initiates the silencing of release sites during depression at sensory neuron-to-motor neuron synapse in Aplysia
In the absence of Ca²⁺ influx no detectable synaptic depressin developed. Thus the switching off of sensory neuron release sites during repetitive presynaptic activity appears to be initiated by presynaptic Ca²⁺ influx.

6. The switch that silences sensory neuron-to-motor neuron synapses during depression in Aplysia involves the small G protein Arf
One possibility is that because facilitation of depressed synapse involves protein kinase C (Ghirardi et al., 1992; Manseau et al., 2001), the switching of synapses to a silent state is mediated by a protein phosphatase. Indeed, some types of long-term depression involve protein phosphatases (...).

... a general inhibitor of Arf signaling powerfuly inhibited transmission at sensory neuron-to-motor neuron synapses and occluded the development of synaptic depression. Similarly, a binding domain of an Arf effector protein acted as a dominant negative and substantially reduced the depression that developed with single action pontentials. Reciprocally, we found that constitutively active Arf6, when injected into presynaptic sensory neurons, prevented the development of synaptic depression.

7. Bursts of sensory neuron activity prevent the development of homosynaptic depression"burst-dependent protection" from synaptic depression
Burst-dependent protection from synaptic depression involves Ca²⁺-dependent activation of protein kinase C (PKC). Chelating Ca²⁺ in the presynaptic sensory neuron with intracellular EGTA, which is not a sufficiently rapid buffer to block transmitter release, eliminates burst-dependent protection.

The recovery paper

My latest paper will appear in the Journal of Neuroscience.
October 21, 2009 | Volume 29 | Number 42 |

They will introduce my paper in "This Week in The Journal"
Development/Plasticity/Repair - "Nerve Transection Induces Circuit Reorganization in Tritonia"

The article title is:
"Functional Recovery after Lesion of a Central Pattern Generator"
by Akira Sakurai and Paul S. Katz

In this paper, we found that severing a set of connections between some CPG neurons impaired motor pattern production but that the system spontaneously recovered over the course of a few hours to a day. Furthermore, we observed corresponding changes in synaptic strength that can account for the functional recovery.

Neuromodulation of motor systems

Among many reviews about CPG funciton, I like this one by Ole Kiehn and Paul S. Katz.
The introduction starts with a brief talk about dancing.

"...by changing cellular and synaptic properties, neuromodulators choreograph circuits from an ensemble of interacting neurons capable f dancing with a variety of partners."

2. The elements of neuromodulation in motor systems: alterations of cellular and synaptic properties
The CPGs - Localized neuronal networks in the central nervous system control the timing of the coordinated muscle activities, capable of producing rhythmic movement even when isolated from the sensory input (Delcomyn 1980).

The CPG function depends on synaptic interconnections and intrinsic membrane properties.
Neuromodulation changes both of them.

1) Rewiring circuits
We may often think of the nervous system as a hard-wired device whose connectivity is changed only during the developental period or as a result of learning.
NO, the strength of connections between neurons is not fixed, but can vary continusously under moment-to-moment neuromodulatory control.
The wiring diagram for a circuit is merely an outline of potential connections and does not uniquely determine the flow of information at all times.
- Modulation of chemical transmission
The effect of neuromodulation can be a functional disconnection of cells or a strengthening of the communication between cells.
Thus, the wiring diagram of synaptic connections is strongly dependent upon which neuromodulator is present.
In vertebrate locomotion, 5-HT and noradrenaline modulate glycinergic synapses to increase circuit flexibility.
- Modulation of electrical coupling

2) Changing neuronal personalities

- Modulation of resting conductances can determine neuronal participation in a network

- Modulation of conductances involved in phase transisions

- Modulation of conductances that determine spike rate

- Modulation of conductances underlying neuronal bistability

- Modulation of conductances underlying conditional bursting


3) Changes in cellular and synaptic properties produce secondary effects

The differential actions of neuromodulators on neurons in motor circuits underlie some forms of behavioral plasticity such as motor pattern selection.


3. Choreographing motor patterns: the effects of neuromodulators on the output of motor circuits

Neuromodulatory substances can initiate motor patterns by endowing neurons with the properties that are needed to form a functional CPG circuit.
Neuromodulatory sunstances can alter (or reorganize) motor patterns by changing those properties.

1) Neuromodulators can activate motor patterns
As a rule, the initiation of rhythmic movements requires non-rhythmic input from a source external to the CPG network itself.
-fast synaptic input (tadpole escape)
-neuromodulatory input (Tritonia swim, cats, rats, rabbits)

2) Neuromodulators can alter ongoing motor activity
- changing the speed/frequency
- muscle force
- phase relationship

3) Neuromodulators can reconfigure networks
-stomatogastric system
- At the moment, little is known about these types of network reorganizations in CPGs other than those in the stomatogastric system. Reconfiguration in the larger neuronal networks that control thythmic activity in vertebrates is difficult to evaluate because the CPG networks are poorly difined and it is impossible to be sure that one has recorded from all possible members of a functional circuit.

4) Neuromodulation can alter the ability of a CPG to drive its follower motor neurons

4. INtegrating neuromodulation into neuronal circuits

1) Properties of neuromodulatory neurons

2) Sources of neuromodulation
- Extrinsic vs Intrinsic

3) Convergence of modulation

5. Long-term alteration of motor patterns
Fast proprioceptive adjustment mechanisms are plastic and that they can adjust to long-term changes in the sensory signaling.

- in spinalized cats where locomotion on a treadmill is evoked by L-DOPA injection, cutting the lateral-gastrocnemius-soleus nerve results in long-term up-regulation of the load-compensating effects from group I afferents in the synergistic medial-gastrocnemius nerve, allowing the cat to slowly recover its normal stepping behavior (Whelan and Pearson 1997).

Neuromodulatory inputs may play a role in promoting long-term plasticity of CPG circuits. In spinalized cats, daily intraperitoneal or intrathecal injections of the alpha-2 adrenergic receptor agonist, clonidine, enhanced the recovery of locomotion when combined with training on a treadmill (Chau et al., 1998).

Melibe leonina

I am working on Melibe brain now. It is so small.

There are two small ganglia on Pleural-Pleural connective nerve. What they do?
A copepod is sitting on the brain in the dish. I think it came out from the stomach, but it is still alive. It is so pity that I have to kill it to protect my prep.

I saw a pair of green tube that seem to go to the dorsal gill from the stomach.
Is it also solar-powered?