Excitatory postsynaptic calcium transients at Aplysia sensory–motor neuron synapses allow for quantal examination of synaptic strength over multiple days in culture

A more thorough description of the changes in synaptic strength underlying synaptic plasticity may be achieved with quantal resolution measurements at individual synaptic sites. Here, we demonstrate that by using a membrane targeted genetic calcium sensor, we can measure quantal synaptic events at the individual synaptic sites of Aplysia sensory neuron to motor neuron synaptic connections. These results show that synaptic strength is not evenly distributed between all contacts in these cultures, but dominated by multiquantal sites of synaptic contact, likely clusters of individual synaptic sites. Surprisingly, most synaptic contacts were not found opposite presynaptic varicosities, but instead at areas of pre- and postsynaptic contact with no visible thickening of membranes. The release probability, quantal size, and quantal content can be measured over days at individual synaptic contacts using this technique. Homosynaptic depression was accompanied by a reduction in release site probability, with no evidence of individual synaptic site silencing over the course of depression. This technique shows promise in being able to address outstanding questions in this system, including determining the synaptic changes that maintain long-term alterations in synaptic strength that underlie memory.

Postactivation Depression of the Soleus H Reflex Measured Using Threshold Tracking

The interpretation of changes in the soleus H reflex is problematic in the face of reflex gain changes, a nonlinear input/output relationship for the motoneuron pool, and a nonhomogeneous response of different motoneurons to afferent inputs. By altering the stimulus intensity to maintain a constant reflex output, threshold tracking allows a relatively constant population of α-motoneurons to be studied. This approach was used to examine postactivation (“homosynaptic”) depression of the H reflex (HD) in 23 neurologically healthy subjects. The H reflex was elicited by tibial nerve stimulation at 0.05, 0.1, 0.3, 1, and 2 Hz at rest and during voluntary plantar flexion at 2.5, 5, and 10% of maximum. A computerized threshold tracking procedure was used to set the current needed to generate a target H reflex 10% of Mmax. The current needed to produce the target reflex increased with stimulus rate but not significantly beyond 1 Hz. In three subjects, the current needed to produce H reflexes of 5, 10, 15, and 20% Mmax at 0.3, 1, and 2 Hz increased with rate and with the size of the test H reflex. HD was significantly reduced during voluntary contractions. Using threshold tracking, HD was maximal at lower frequencies than previously emphasized, probably because HD is greater the larger the test H reflex. This would reinforce the greater sensitivity of small motoneurons to reflex inputs.