Significance: The standard view of neural signaling is that a neuron can influence its target cell by exciting or inhibiting it. An important aspect of the standard view is that learning consists of changing the efficacy of synapses, either strengthening (long-term potentiation) or weakening (long-term depression) them. In studying how cerebellar Purkinje cells change their responsiveness to a stimulus during learning of conditioned responses, we have found that these cells can learn the temporal relationship between two paired stimuli. The cells learn to respond at a particular time that reflects the time between the stimuli. This finding radically changes current views of both neural signaling and learning.

Cerebellar Purkinje cells can learn to respond to a conditioned stimulus with an adaptively timed pause in firing. This response was usually ascribed to long-term depression of parallel fiber to Purkinje cell synapses but has recently been shown to be due to a previously unknown form of learning involving an intrinsic cellular timing mechanism. Here, we investigate how these responses are elicited. They are resistant to blockade of GABAergic inhibition, suggesting that they are caused by glutamate release rather than by a changed balance between GABA and glutamate. We show that the responses are abolished by antagonists of the mGlu7 receptor but not significantly affected by other glutamate antagonists. These results support the existence of a distinct learning mechanism, different from changes in synaptic strength. They also demonstrate in vivo post-synaptic inhibition mediated by glutamate and show that the mGlu7 receptor is involved in activating intrinsic temporal memory.

Cerebellar learning requires context information from mossy fibers and a teaching signal through the climbing fibers from the inferior olive. Although the inferior olive fires in bursts, virtually all studies have used a teaching signal consisting of a single pulse. Following a number of failed attempts to induce cerebellar learning in decerebrate ferrets with a nonburst signal, we tested the effect of varying the number of pulses in the climbing fiber teaching signal. The results show that training with a single pulse in a conditioning paradigmin vivodoes not result in learning, but rather causes extinction of a previously learned response.

Significance: Learning is thought to rely on the strengthening or weakening of synapses. However, we have previously shown that a neuron can also learn when to time its response so that the timing reflects the interval between two stimuli, without any temporal information in the input signal. Here, we report that a neuron can even learn a sequence of at least two, and probably more, accurately timed responses. A single cell is in a sense “programmable,” and can encode a temporal response pattern. This means that the nature of what a cell can learn is very different from the traditional view, and that the information storage capacity may be far greater.

Many forms of learning require temporally ordered stimuli. In Pavlovian eyeblink conditioning, a conditioned stimulus (CS) must precede the unconditioned stimulus (US) by at least about 100 ms for learning to occur. Conditioned responses are learned and generated by the cerebellum. Recordings from the cerebellar cortex during conditioning have revealed CS-triggered pauses in the firing of Purkinje cells that likely drive the conditioned blinks. The predominant view of the learning mechanism in conditioning is that long-term depression (LTD) at parallel fiber (PF)–Purkinje cell synapses underlies the Purkinje cell pauses. This raises a serious conceptual challenge because LTD is most effectively induced at short CS–US intervals, which do not support acquisition of eyeblinks. To resolve this discrepancy, we recorded Purkinje cells during conditioning with short or long CS–US intervals. Decerebrated ferrets trained with CS–US intervals ≥150 ms reliably developed Purkinje cell pauses, but training with an interval of 50 ms unexpectedly induced increases in CS-evoked spiking. This bidirectional modulation of Purkinje cell activity offers a basis for the requirement of a minimum CS–US interval for conditioning, but we argue that it cannot be fully explained by LTD, even when previousin vitrostudies of stimulus-timing-dependent LTD are taken into account.