Time course of the effects of injection
of ApCREB2 antiserum on short- and long-term facilitation.
(A) Time course of excitatory postsynaptic potenial (EPSP)
amplitude changes recorded in motor neuron L7 in response to
stimulation of the sensory neuron (expressed as percent change in the
amplitude of the EPSP) after single and multiple applications of 5-HT
to Aplysia sensory–motor neuron cocultures. Changes in
EPSP amplitude after application of one 5-min pulse of 5-HT (1× 5-HT,
short-term facilitation) and one 5-min pulse of 5-HT paired with
injection of anti-ApCREB2 antibodies (1× 5-HT + CREB-2 Ab, both in
boldface lines) are compared with changes in EPSP amplitude induced by
five pulses of 5-HT (5× 5-HT) at 2 and 24 hr. While the EPSP
facilitation decays rapidly after one pulse of 5-HT (with a return to
base line after 10 min), pairing one pulse of 5-HT with injection of
anti-ApCREB2 antibodies induces a long-term
facilitation paralleling that of 5× 5-HT. This long-term facilitation
is abolished by the application of the protein synthesis inhibitor
anisomycin (1× 5-HT + CREB-2 Ab + ANISO) or the RNA synthesis
inhibitor actinomycin D (1× 5-HT + CREB-2 Ab + ACTINO) during the
training. The difference in EPSP amplitude at 2 hr between 5× 5-HT and
1× 5-HT + CREB-2 Ab may reflect the transient protein
synthesis-dependent, but RNA synthesis-independent, component of
long-term facilitation 2 hr after 5-HT stimulation (29). The controls
are either untreated (control), or injected with ApCREB2 antiserum
without 5-HT administration (CREB-2 Ab). (B) Comparison of
the time course of the EPSP amplitude changes in the first 2 hr after
application of a single 5-min pulse of 5-HT with or without injection
of CREB-2 antibody. The control cells were not exposed to 5-HT.
(C) Example of EPSPs recorded in motoneuron L7 after
stimulation of the sensory neuron before (0 hr) and 2 and 24 hr after
5-HT treatment. One pulse of 5-HT paired with the injection of an
ApCREB2 antiserum induces a significant increase in EPSP amplitude at 2
and 24 hr, but injection the preimmune serum (PRE-CREB-2 Ab) or
depleted immune serum does not induce long-term facilitation.
(D) Examples of EPSPs recorded at indicated times in
cocultures injected with ApCREB2 antiserum paired with one 5-min pulse
of 5-HT. [Reproduced with permission from Bartsch et
al. (27) (Copyright 1995, Cell Press).]