Signal Transduction Mechanisms and Early Events
Signal Transduction Mechanisms and Early Events
Scientists believe that long-lasting changes in synaptic function are essential for learning and memory in the mammalian brain. A widely studied example of such synaptic plasticity is long-term potentiation (LTP). The remarkable feature of LTP is that a short burst of synaptic activity can trigger persistent enhancement of synaptic transmission lasting for at least several hours, and possibly weeks or longer. There is great interest in understanding the cellular and molecular mechanisms that underlie this form of synaptic plasticity. First found in the hippocampus, this phenomenon is now known to exist in cerebral cortex and other areas of the mammalian central nervous system (CNS). Indeed, damage to the hippocampus can result in certain defects in memory acquisition (see Milner, Squire, and Kandel, 1998).
Most studies on LTP focus on the synapse between Schaffer collaterals and hippocampal CA1 neurons. In this system, a brief burst of afferent stimulation leads to induction of LTP in postsynaptic CA1 cells through a combination of (1) membrane depolarization and (2) activation of glutamate receptors of the NMDA subtype (e.g., Collingridge, Kehl, and McLennan, 1983). Researchers generally agree that the depolarization relieves Mg2+ (magnesium ion) block of NMDA receptor channels and allows a Ca2+(calcium ion) influx into dendritic spines that somehow triggers LTP (Nicoll, Kauer, and Malenka, 1988).
Back-Propagating Action Potentials
In hippocampal pyramidal neurons an important component of the membrane depolarization that allows opening of NMDA receptors is back-propagating action potentials. While action potentials are of course triggered in the active zone of the cell body, hippocampal pyramidal neurons along with many other types of CNS neurons can actively propagate action potentials into the dendritic regions. These dendritic action potentials are just like action potentials propagated down axons in that they are carried predominantly by voltage-dependent ion channels such as sodium channels. The penetration of the back-propagating action potential into the dendritic region provides a wave of membrane depolarization that allows for the opening of the voltage-dependent NMDA receptor/ion channels. In fact, the timing of the arrival of a dendritic action potential with synaptic glutamate input appears to play an important part in precise, timing-dependent triggering of synaptic plasticity in the hippocampus (Magee and Johnston, 1997). Moreover, modulatory neurotransmitter systems can regulate the likelihood of action potential back-propagation through controlling dendritic potassium channels, allowing for sophisticated information processing through an interplay of action potential propagation, glutamate release, and neuromodulation (Johnston, Hoffman, Colbert, and Magee, 1999).
Importance of a Rise in Postsynaptic [Ca2+]i
NMDA receptor activation leads to a transient [Ca2+]i increase arising in the postsynaptic neuron, an effect that researchers have measured by the use of fluorescent Ca2+ indicator dyes in pyramidal cell dendrites within hippocampal slices (Regehr and Tank, 1990). A variety of experiments have demonstrated the importance of the rise in [Ca2+]i for LTP. Ca2+ buffers such as EGTA or BAPTA have been introduced with the aim of suppressing the transient Ca2+ increase; such maneuvers are effective in preventing the induction of LTP (Lynch et al., 1983). Moreover, a rise in postsynaptic Ca2+. independent of glutamate receptors, has been imposed by photoactivation of a caged Ca2+ compound, nitr-5; this method for Ca2+ delivery causes a sustained synaptic potentiation (Malenka, Kauer, Zucker, and Nicoll, 1988).
Involvement of Protein Kinases
The key question at this point is how a relatively brief rise in [Ca2+]i can lead to a long-lasting enhancement of synaptic function. One popular hypothesis is that Ca2+ acts by activating signal transduction pathways sensitive to the elevation of postsynaptic Ca2+. The pathways that have been implicated are quite varied and include: the Ca2+-phospholipid dependent protein kinase (protein kinase C, PKC), multifunctional Ca/calmodulin-dependent protein kinase (CaMKII), calcium/calmodulin sensitive adenylyl cyclase and the PKA pathway, the ras/ERK MAP kinase pathway, and calcium-responsive nitric oxide (NO) synthase (Adams and Sweatt, 2002; Lisman and Zhabotinsky, 2001; Lu, Kandel, and Hawkins, 1999; Sweatt, 1999; Hrabetova and Sacktor, 1996).
Involvement of Ca2+ -dependent protein kinases has been extensively tested and a wide variety of evidence is compatible with the general hypothesis of a necessity for postsynaptic protein kinase activation as being necessary for triggering LTP (see Figure 1). A necessity for PKC and CaMKII has been tested by intracellular injection of peptides that are potent and selective inhibitors of either PKC or CaMKII (Malinow, Schulman, and Tsien, 1989), and by characterization of kinase-deficient mice generated through gene knockout technology (Abielovich et al., 1993; Silva, Stevens, Tonegawa, and Wang, 1992). Pharmacologic inhibitors of MAP kinase activation have also been shown to block LTP induction (Adams and Sweatt, 2002). Involvement of the PKA pathway has been probed using pharmacologic and transgenic animal approaches, as has the involvement of the NO synthase/cGMP pathway (Lu, Kandel, and Hawkins, 1999; and Sweatt, 1999).
The results obtained with inhibition of PKC, CaMKII, PKA, MAP Kinase, and the NO/cGMP pathway can be interpreted in terms of a network of protein kinases, with protein phosphorylation as a link between the rise in [Ca2+]i and the eventual expression of enhanced synaptic function. However, the topology of the network and the nature of the interactions remain undefined. Indeed, there is no evidence to date to exclude the idea that one or more of these enzymes act in a merely permissive way. At one extreme, background activity of a particular kinase might be necessary only prior to induction, to set the stage for some other signaling mechanism triggered by Ca2+.
Evidence for Presynaptic Expression of LTP
The question of how the kinases act leads to consideration of ongoing debate about the nature of the maintained synaptic enhancement in LTP. There is a variety of evidence to support the view that both increased presynaptic transmitter release and enhanced glutamate receptor function postsynaptically are involved. The results indicating enhancement presynaptically support the idea of a retrograde signal that travels from the postsynaptic cell back to the presynaptic terminal. This line of reasoning has led to a search for specific compounds that might act as the retrograde messenger, with arachidonic acid, nitric oxide, and superoxide being the most widely considered possibilities (Sweatt, 1999). The precise mechanisms by which these retrograde signals might affect neurotransmitter release are unclear, although presynaptic activation of PKC and the cGMP-dependent protein kinase are appealing possibilities.
Little is known for certain about possible pre-synaptic mechanisms that might be set in motion by putative retrograde messengers. A widely considered mechanism for synaptic potentiation involves a persistent enhancement of a presynaptic protein kinase, such as PKC (Linden and Routtenberg, 1989). The maintained expression of LTP can be reversibly blocked by bath application of a relatively nonspecific kinase inhibitor, H-7 (Malinow, Madison, and Tsien, 1988), and biochemical measurements show a sustained enhancement of PKC in hippocampal slices (Klann, Chen, and Sweatt, 1993; Hrabetova and Sacktor, 1996). PKC is known to increase the efficiency of excitation-secretion coupling in many systems, including chromaffin cells, motor nerve terminals, and cultured hippocampal neurons. Further exploration of presynaptic mechanisms is needed to determine which steps leading to exocytosis are enhanced in LTP.
Mechanisms of Postsynaptic Enhancement
In terms of mechanisms for enhancing postsynaptic responsiveness, two types of mechanisms are being actively pursued experimentally. One type of process is referred to as activation of "silent synapses" (Poncer and Malinow, 2001; Isaac, Nicoll, and Malenka, 1999). The concept is that some latent synapses have available only NMDA receptors and thus are inactive in terms of normal baseline synaptic transmission. Activation of NMDA receptors by the mechanisms described above leads to calcium influx and calcium-mediated insertion of AMPA receptors locally. Thus previously "silent" synapses become capable of baseline synaptic transmission and the net result is an increased efficacy of transmission.
A second mechanism under investigation is kinase-dependent regulation of AMPA receptors through direct phosphorylation. It is known that phosphorylation of AMPA receptors by PKC and CaMKII leads to increased conductance of ions through the AMPA channel, and evidence is available that phosphorylation at these regulatory sites increases with LTP-inducing stimulation (Lee et al., 2000; Derkach, Barria, and Soderling, 1999; Barria et al., 1997). Thus it is hypothesized that through increased postsynaptic phosphorylation of AMPA receptors synaptic efficacy is increased by enhancement of AMPA receptor function. The "silent synapse" hypothesis and the enhanced AMPA receptor conductance hypothesis are not mutually exclusive; indeed both could be simultaneously occurring at different synapses between the same cells, or at the same synapses at different points in time.
Potential Roles for Protein Phosphatase Regulation
Finally, while increased protein phosphorylation has been hypothesized to play a key role in triggering LTP for quite some time, studies conducted since the turn of the century highlight a similar importance of tightly regulating protein dephosphorylation as one of the mechanisms controlling the induction of long-term synaptic change and lasting memory (Winder and Sweatt, 2001). In one series of studies, Isabel Mansui and her colleagues (Mallaret et al., 2001) engineered a mouse line in which the activity of the calcium-dependent phosphatase calcineurin (protein phosphatase 2B) could be regulated, in a reversible manner, by tetracycline administration to adult animals. In engineering their mouse they capitalized on the known mechanism of regulation of calcineurin; that is, its regulation by an intrinsic autoinhibitory domain. Mansui's group engineered a mouse in which they could regulate the expression of the isolated autoinhibitory domain of the calcineurin A α subunit. In their mice, treatment of animals with tetracycline led to expression of the calcineurin autoinhibitory domain protein in several brain regions, including the hippocampus. They then investigated long-term potentiation in hippocampal slices, specifically in area CA1 where many of the earlier studies implicating kinases had been performed. Calcineurin inhibition enhanced the magnitude and duration of long-term potentiation in this region when LTP was triggered by high-frequency stimulation. These studies and a variety of others (Winder and Sweatt, 2001) suggest the possibility of an important role for calcium-dependent phosphatases in regulating the magnitude of LTP induction in the hippocampus.
Since the early 1990s, much progress has been made in scientific understanding of the mechanisms involved in LTP induction. A role for protein phosphorylation in LTP induction is clear. Dendritic action potentials were inconceivable in the early 1990s but in the twenty-first century are known to be critical components of information processing in neurons, through their contributions to LTP induction. Many questions remain concerning the targets of protein kinases in LTP, the role of morphological changes and local protein synthesis in early stages of LTP, and the seemingly perpetual question of pre-versus postsynaptic loci of LTP expression. Scientists are hopeful that continued research and progress will shed light on these topics.
See also:GLUTAMATE RECEPTORS AND THEIR CHARACTERIZATION; GUIDE TO THE ANATOMY OF THE BRAIN: HIPPOCAMPUS AND PARAHIPPOCAMPAL REGION; LONG-TERM POTENTIATION: OVERVIEW: COOPERATIVITY AND ASSOCIATIVITY; SECOND MESSENGER SYSTEMS
Abeliovich, A., Chen, C., Goda, Y., Silva, A. J., Stevens, C. F., and Tonegawa, S. (1993). Modified hippocampal long-term potentiation in PKC gamma-mutant mice. Cell 75 (7), 1,253-1,262.
Adams, J. P., and Sweatt, J. D. (2002). Molecular psychology: Roles for the ERK MAP kinase cascade in memory. Annual Review of Pharmacology and Toxicology 42, 135-163.
Barria, A., Muller, D., Derkach, V., Griffith, L. C., and Soderling, T. R. (1997). Regulatory phosphorylation of AMPA-type glutamate receptors by CaM-KII during long-term potentiation. Science 276 (5,321), 2,042-2,045.
Collingridge, G. L., Kehl, S. J., and McLennan, H. (1983). Excitatory amino acids in synaptic transmission in the Schaffer collateral-commissural pathway of the rat hippocampus. Journal of Physiology 334, 33-46.
Derkach, V., Barria, A., and Soderling, T. R. (1999). Ca2+/calmodulin-kinase II enhances channel conductance of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate type glutamate receptors. Proceedings of the National Academy of Sciences of the United States of America 96 (6), 3,269-3,274.
Hrabetova, S., and Sacktor, T. C. (1996). Bidirectional regulation of protein kinase M zeta in the maintenance of long-term potentiation and long-term depression. Journal of Neuroscience 16 (17), 5,324-5,333.
Isaac, J. T., Nicoll, R. A., and Malenka, R. C. (1999). Silent glutamatergic synapses in the mammalian brain. Canadian Journal of Physiology and Pharmacology 77 (9), 735-737.
Johnston D., Hoffman, D. A., Colbert, C. M., and Magee J. C. (1999). Regulation of back-propagating action potentials in hippocampal neurons. Current Opinion in Neurobiology 9 (3), 288-292.
Klann, E., Chen, S.-J., and Sweatt, J. D. (1993). Mechanism of PKC activation during the induction and maintenance of LTP probed using a novel peptide substrate. Proceedings of the National Academy of Sciences of the United States of America 90, 8,337-8,341.
Lee, H. K., Barbarosie, M., Kameyama, K., Bear, M. F., and Huganir, R. L. (2000). Regulation of distinct AMPA receptor phosphorylation sites during bidirectional synaptic plasticity. Nature 405 (6,789), 955-959.
Linden, D. J., and Routtenberg, A. (1989). The role of protein kinase C in long-term potentiation: A testable model. Brain Research Reviews 14, 279-296.
Lisman, J. E., and Zhabotinsky, A. M. (2001). A model of synaptic memory: A CaMKII/PP1 switch that potentiates transmission by organizing an AMPA receptor anchoring assembly. Neuron 31 (2), 191-201.
Lu, Y. F., Kandel, E. R., and Hawkins, R. D. (1999). Nitric oxide signaling contributes to late-phase LTP and CREB phosphorylation in the hippocampus. Journal of Neuroscience 19 (23), 10,250-10,261.
Lynch, G., Larson, J., Kelso, S., Barrionuevo, G., and Schottler, F. (1983). Intracellular injections of EGTA block induction of hippocampal long-term potentiation. Nature 305, 719-721.
Magee, J. C., and Johnston, D. (1997). A synaptically controlled, associative signal for Hebbian plasticity in hippocampal neurons. Science 275 (5,297), 209-213.
Malenka, R. C., Kauer, J. A., Zucker, R. S., and Nicoll, R. A. (1988). Postsynaptic calcium is sufficient for potentiation of hippocampal synaptic transmission. Science 242, 81-84.
Malinow, R., Madison, D. V., and Tsien, R. W. (1988). Persistent protein kinase activity underlying long-term potentiation. Nature 335, 820-824.
Malinow, R., Schulman, H., and Tsien, R. W. (1989). Inhibition of postsynaptic PKC or CaMKII blocks induction but not expression of LTP. Science 245, 862-866.
Malleret, G., Haditsch, U., Genoux, D., Jones, M. W., Bliss, T. V., Vanhoose, A. M., Weitlauf, C., Kandel, E. R., Winder, D. G., and Mansuy, I. M. (2001). Inducible and reversible enhancement of learning, memory, and long-term potentiation by genetic inhibition of calcineurin. Cell 104, 675-686.
Milner, B., Squire, L. R., and Kandel, E. R. (1998). Cognitive neuroscience and the study of memory. Neuron 20 (3), 445-468.
Nicoll, R. A., Kauer, J. A., and Malenka, R. C. (1988). The current excitement in long-term potentiation. Neuron 1, 97-103.
Poncer, J. C., and Malinow, R. (2001). Postsynaptic conversion of silent synapses during LTP affects synaptic gain and transmission dynamics. Nature Neuroscience 4 (10), 989-996.
Regehr, W. G., and Tank, D. W. (1990). Postsynaptic NMDA receptor-mediated calcium accumulation in hippocampal CA1 pyramidal cell dendrites. Nature 345, 807-810.
Silva, A. J., Stevens, C. F., Tonegawa, S., and Wang, Y. (1992). Deficient hippocampal long-term potentiation in alpha-calciumcalmodulin kinase II mutant mice. Science 257 (5,067), 201-206.
Sweatt, J. D. (1999). Toward a molecular explanation for long-term potentiation. Learning and Memory 6, 399-416.
Winder, D. G., and Sweatt, J. D. (2001). Roles of serine/threonine phosphatases in hippocampal synaptic plasticity. Nature Reviews Neuroscience 2, 461-474.
Revised byJ. DavidSweatt