Using more physiological stimuli like so called theta bursts

(1 TBS = 4 pulses at 100 Hz separated by 200 ms repeated 12 times):

three forms of LTP with different duration and mechanisms at CA3-CA1 synapses:

LTP1 (< 2 hours, 1 TBS) depends on modifications of postsynaptic proteins (mainly

by CaMKII, not PKC); depends also on Ca release from intraspine Ca stores via
RyR (Ca-induced Ca release).

LTP2 (4-5 hours, 4 TBS) requires new protein synthesis from preexisting mRNA
with translation machinery located in the dendrites, and depends on coactivation of
NMDARs and mGluRs, leading to activation of PKC

LTP3 (>= 24 hours, 8 TBS) requires both gene transcription and protein synthesis.
It develops slowly. It depends on Ca influx through CaV1.2 channels (not on
NMDAR and Ca release from stores) and on activation of MAP-kinase which
translocates in nucleus in PKA-dependent manner.
Activation of ras-MAPkinase pathway by Ca influx through CaV1.2 channels in
LTP3 leads to sustained CREB phosphorylation, in contrast with transient
phosphorylation produced by activation of the same pathway by Ca influx through
NMDARs
 PKC is activated as a
consequence of
coactivation of NMDARs
and mGluRs by the LTP-
inducing stimulus. Proteic
synthesis in dendrites is
regulated by the ERK-MAP
kinase pathway whose
activation depends on PKC.

Calcium entry via

CaV1.2 has a
priviliged role in
activation of gene Activation of PKC-
transcription (in dependent ERK-MAPK
particular CRE- pathway occurs
dependent) in independently from
response to activation of PKA-
neuronal activity dependent ERK-MAPK
pathway, due to
compartmentalization of
Ca2+ signals
Specific patterns of gene
transcription are selectively
induced by Ca2+ influx
through specific Ca2+ -
permeable channels.

Sustained phosphorylation
of CREB (and sustained
gene transcription) due to
activation of Ras-MAPK
pathway in LTP3 depend on
Ca2+ influx through CaV1.2
channels (not NMDAR
channels whose activation
The translocation in the
leads to transient
nucleus of MAPK is PKA-
phosphorylation of CREB,
dependent.
due to simultaneous
Activation of PKA depends on
activation of PP1).
coactivation of dopaminergic
and noradrenergic
metabotropic receptors

Nanou and Catterall (2018) Neuron

 Synaptic tagging

Synapse-specificity implies that signals are not only sent from synapse to nucleus but
also from nucleus to synapse, i.e.newly synthetized gene products, both mRNA and
proteins, have to be delivered to the specific synapses whose activation originally
triggered the gene expression.
 Synaptic tagging hypothesis: products of gene expression are delivered throughout
the cell, but are only used at synapses that have been tagged by previous activity.

Cf Frey and Morris (1997 Nature and Bear’s News and views for nice exp showing that
LTP3 and its associated synaptic changes are synapse specific).

How is an active synapse marked? What is the synapse-specific tag?

• a regulator of gene translation that activates protein synthesis at the synapse (cytoplasmic polyadenilation element
binding protein CPEB in Aplysia, with N terminus resembling prion domain endowing CPEB with similar self-
sustaining properties. At an inactive synapse CPEB is inactive or repressed; the active prion-like state is
established only at strongly activated synapses; once prion state is established, mRNA molecules made in the cell
body and distributed throughout the cell are translated but only at that activated synapse. Mouse homolog CPEB3,
which is activated by neuralized1, an E3 ubiquitin ligase).
• PKA-dependent phosphoprotein?
 Key role of NMDAR, as at the CA3-CA1
synapse.
Induction of LTP at DG-CA3 synapses
depends on Ca influx at the presynaptic
terminals that activates a CaM-dependent
adenylate cyclase, increases cAMP,
activates PKA and increases AP-evoked
glutamate release.

Non associative LTP, indipendent from depolarization of postsynaptic cell

 Long term depression (LTD)
at CA3-CA1 hippocampal synapses
LTD develops following low frequency
stimulation (1 Hz) for a long period (10-
15 min).

LTD can eliminate the increase in EPSP

amplitude produced by LTP (and
viceversa).
This suggests the involvement of
modulation at a common site.

Also induction of LTP depends on

activation of NMDARs and Ca influx into
the spine, but the key difference seems
to be the extent of Ca2+ increase: in LTD
Mg2+ block is removed in only a few
NMDARs and there is a small Ca2+
increase sufficient to activate Ca-
dependent phosphatases (and
consequent activation of other
phosphatases) but not CaMKII (or other
Ca-dependent kinases).
The expression of LTD depends on
internalization of AMPA receptors

NB also for LTD, the underlying mechanisms can be different at different synapses
Despite the low frequency of their discharge,
climbing fibers modulate the input of parallel
fibers to Purkinje cells. In particular, climbing
fibers can selectively induce long-term
depression in the synapses between parallel
fibers and Purkinje neurons that are activated
concurrently with the climbing fibers.

Concurrent stimulation of climbing fibers

and parallel fibers depresses the Purkinje
cell responses to subsequent stimulation
of the same parallel fibers but not to
stimulation of parallel fibers that had not
been stimulated earlier along with
climbing fibers. The resulting
depression can last for minutes to hours.
What functional effects might this long-term
depression have?
According to the theories of Marr, Albus and
Ito, altering the strength of certain synapses
between parallel fibers and Purkinje cells
shapes or corrects eye and limb movements.
During an inaccurate movement the climbing
fibers respond to specific movement errors and
depress the synaptic strength of parallel fibers
involved with those errors, namely those that
had been activated with the climbing fiber. With
successive movements the parallel fiber inputs
conveying the flawed central command
are increasingly suppressed, a more
appropriate pattern of simple-spike activity
emerges, and eventually the climbing-fiber
error signal disappears
 LTD at the synapses between parallel fibers and Purkinje cells induced by
simultaneous activation of parallel fibers and climbing fibers:

Climbing fiber Parallel fiber

Weakened synapse

Climbing fiber activation

depolarizesmembrane
and open CaV2.1

Simultaneous activation of CaV2.1

channels (due to large EPSP produced
by climbing fiber activation) and of
mGluRs at PF-PC synapses (resulting in
IP3 and DAG production) leads to release
of Ca2+ from intracellular stores and
synergistic activation of PKC by Ca and
DAG. At this synapses proteins
phosphorylated by PKC leads to
internalization of AMPA receptors and
LTD.

Note that here the coincidence detectors

are the IP3Rs and PKC
 STPD: spike timing-dependent plasticity:
a form of LTP induced by pairing of a single presynaptic AP with the firing of a single AP in the postsynaptic cell
Presynaptic AP before
postsynaptc AP EPSC (% control)
The pairing protocol produces
LTP only if the postsynaptic cell
Postsynaptic V
fires a few ms after the
beginning of the EPSP: in this
case the postsynaptic AP
relieves the Mg block at a time
when the NMDAR has been
Presynaptic AP after
opened by the binding of
postsynaptc AP glutamate (but inconsistent with
time window: other mechanisms
Postsynaptic V
are involved).

Different mechanisms (still

unclear) at different synapses

Pre before
Post before
post
The two APs need to be
pre
EPSC (% control) separated by >40 ms

Important role in cerebral processing

:
Feldman (2012) Neuron

Interval
Interval between
between presynaptic
presynaptic and
AP and postsynaptc AP
Neural circuits can represent many different external events and encode
a wide range of memories. It is assumed that any individual neurons can
participate in different representations or memories.
How can a new memory be formed through altered synaptic strength
without overwriting a preexisting memory encoded in a neuron’s
synapses?

Kandel et al (2014) Cell

The molecolar and systems biology of memory
 Kandel et al (2014) Cell:
The molecular and systems biology of memory

Tonegawa et al (2015) Curr Opin Neurobiology

Memory engram storage and retrieval

Using c-fos based genetic tagging approach, ChR2 is expressed specifically in

neurons that were activated during learning in a contextual fear-conditioning
test.
When light pulses were delivered to the dentate gyrus to stimulate the ChR2
expressing neurons the animals showed fear light-induced reactivation of
the cells active during learning results in memory retrieval
These labelled cells were reactived also upon presentation of retrieval cues;
their reactivation can elicit memory recall
They are part of a memory engram ensemble (Liu et al, 2012 Nature; Ramirez
et al, 2013 Science)
Engram cells display changes in synaptic weight typical of LTP (Ryan et al,
2015 Science).