Systems Neurobiology Spring School 2005
Japanese / English
|title||Dissecting the cell cycle oscillator
We have been studying the systems-level design principles of the
Cdc2-APC system, the network of proteins responsible for driving cell
cycle oscillations. The system is driven by alternating periods of
cyclin accumulation (during interphase) and destruction (during
M-phase). The switch between accumulation and destruction is carried
out by a negative feedback loop. In addition, there are multiple
positive feedback loops: active Cdc2 activates its own activator
Cdc25, and active Cdc2 inactivates its inhibitors Wee1 and Myt1. We
have investigated the roles of the positive and negative feedback
loops individually, using biochemically-tractable Xenopus egg
extracts as our experimental system and computational modeling to
guide our experiments.
- Pomerening JR, Sontag ED, Ferrell JE. Building a cell cycle oscillator: hysteresis and bistability in the activation of Cdc2. Nat Cell Biol. 2003 Mar 10.
- Angeli D, Ferrell JE Jr, Sontag ED. Detection of multistability, bifurcations, and hysteresis in a large class of biological positive-feedback systems.
Proc Natl Acad Sci U S A. 2004 Feb 17;101(7):1822-7. Epub 2004 Feb 06.
|title||BDNF and Hippocampal Synaptic Plasticity
While traditionally viewed as secretory growth factors involved in long-term
survival and differentiation of neurons, neurotrophins have now been
recognized as important factors that regulate synaptic transmission and
plasticity, synapse development, and learning and memory. We have previously
demonstrated that brain-derived neurotrophic factor (BDNF) acutely
facilitates hippocampal long-term potentiation (LTP), a cellular model for
learning and memory.
A prevailing phenomenon is that BDNF preferentially modulates active
synapses. To address how diffusible molecules such as BDNF elicit local and
synapse-specific modulation, we have carried out a number of studies on the
cell biology of BDNF and its receptor TrkB. One mechanism is the
activity-dependent control of the trafficking of the TrkB receptors. We
demonstrated that high frequency neuronal activity facilitates the insertion
of TrkB on cell surface. The endocytosis of the BDNF-TrkB complex, which is
critical for certain functions of BDNF, is also regulated by neuronal
activity and BDNF-induced tyrosine kinase phosphorylation of the TrkB
itself. Recently, we have demonstrated that neuronal activity and consequent
elevation of intracellular cAMP concentration control TrkB translocation
into the dendritic spines as well as TrkB tyrosine kinase activity. Thus,
activity control of TrkB receptors is mediated by multiple mechanisms.
The second way by which the synapse-specific BDNF regulation could
be achieved is the activity-dependent secretion of BDNF locally at synapses.
Our recent studies showed that synaptic targeting and activity-dependent
secretion of BDNF may be critical for human memory and hippocampal function.
In the human BDNF gene, there is a single nucleotide polimorphism (SNP) that
convert a valine (val) to a methionin (met) in the 5' pro-region of the BDNF
protein. Human subjects with met allele exhibit lower neuronal activity and
abnormal hippocampal activation, as well as poorer episodic memory. We
showed that in transfected hippocampal cultures met-BDNF cannot be correctly
targeted to the synapses and cannot undergo activity-dependent secretion.
Recently studies indicate that pro-neurotrophins could serve as
signalling molecules by interacting with p75 neurotrophin receptor (p75NTR).
Interestingly, pro-neurotrophins often elicit biological effects that are
opposite to those induced by mature neurotrophins. Thus, protealytic
cleavage of pro-neurotrophins, either intracellularly or extracellularly,
represents a new mechanism that controls the direction of neurotrophin
regulation. In a recent study, we demonstrated that tPA, by activating the
extracellular protease plasmin, converts the precursor proBDNF to mature
BDNF, and that such conversion is critical for L-LTP expression in the
hippocampus 11. There is also evidence that secreted proBDNF, if not
cleaved, binds p75NTR on the dendritic spines of CA1 pyramidal neurons,
leading to an enhancement of NMDA-dependent LTD. Thus, cleavage of proBDNF
(or not) by tPA/plasmin system may serve as an important mechanism for
bi-directional control of hippocampal plasticity: uncleaved proBDNF enhances
LTD through p75NTR, while mature BDNF facilitates LTP though TrkB.
Taken together, our studies provide new insights into the molecular
mechanisms for the synaptic functions of neurotrophins.
- H. Lou, S.K. Kim, E. Zaitsev, C.R. Sneil, B. Lu, and Y.P. Loh (2005) Sorting and activity dependent secretion of BDNF requires an interaction with the sorting receptor Carboxypeptidase E, Neuron 45, 245-255.
- P. T. Pang, H. K. Teng, N. Woo, E. Zaitsev, K. Sakata, S. Zhen, K. K. Teng, W.-H. Yung, B. L. Hempstead, and B. Lu (2004) Cleavage of ProBDNF by the tPA/plasmin is essential for long-term hippocampal plasticity, Science 306, 487-491.
- Y. Ji, P. T. Pang, L. Feng, and B. Lu (InPress) Cyclic AMP Controls BDNF-induced TrkB Phosphorylation and Dendritic Spine Formation in Hippocampal Neurons, Nature Neuroscience.
|title||Distinc Dynamics of transient and sustained ERK activation
To elucidate how epidermal growth factor (EGF) and nerve growth factor
(NGF) specifically encode their distinct physical properties into
transient, and transient and sustained extracellular signal-regulated
kinase (ERK) activation respectively, we developed a kinetic simulation
model of ERK signalling networks by constraining in silico dynamics on
the basis of in vivo dynamics in PC12 cells. Measured in vivo dynamics
can be consistently reproduced in dose- and temporal-dependent manners
in growth factors in silico. This model allowed us to predict in silico
and validate in vivo that transient ERK activation depends on rapid
increases of EGF and NGF but not on their final concentrations, whereas
sustained ERK activation depends on the final concentration of NGF but
not on the temporal rate of increase. This system of ERK dynamics is
produced by Ras and Rap1 dynamics, which capture the rapid temporal rate
of growth factors and the final concentration of NGF, respectively.
These results indicate that the Ras and Rap1 systems capture the
temporal rate and concentration of growth factors, and encode these
distinct physical properties into transient and sustained ERK
- Vaudry, D., Stork, P. J., Lazarovici, P. & Eiden, L. E. Signaling
pathways for PC12 cell differentiation: making the right connections.
Science 296, 1648-1649 (2002).
- Bhalla, U. S. & Iyengar, R. Emergent properties of networks of
biological signaling pathways. Science 283, 381-387 (1999).
Molecular mechanisms of neurite growth: Co-ordinated roles of the
cytoskeleton, cell adhesion molecules, and membrane microenvironment
During development, neurons send out neurites that differentiate into
axons or dendrites. The axons elongate along the correct path to
reach their appropriate targets. The tip of neurites, called the
growth cone, expresses a variety of functional molecules including the
cytoskeleton (actin filaments and microtubules) and cell adhesion
molecules (CAMs). Actin filaments move in a backward direction and
generate traction force that can be transmitted to the extracellular
substrate via CAMs. The growth cone internalizes and recycles CAMs in
a region-specific manner to polarize its adhesivity. Furthermore,
CAM-associated signals could be influenced by the lipid
microenvironment in the cell membrane. In this seminar, I will
demonstrate the importance of dynamic and co-ordinated functions of
these molecules in neurite growth.
- Nishimura K, Yoshihara F, Tojima T, Ooashi N, Yoon W, Mikoshiba K, Bennett V, Kamiguchi H: L1-dependent neuritogenesis involves ankyrinB that mediates L1-CAM coupling with retrograde actin flow. J Cell Biol 163: 1077-1088, 2003.
- Nishimura T, Fukata Y, Kato K, Yamaguchi T, Matsuura Y, Kamiguchi H, Kaibuchi K: CRMP-2 regulates polarized Numb-mediated endocytosis for axon growth. Nat Cell Biol 5: 819-826, 2003.
- Kamiguchi H, Lemmon V: Recycling of the cell adhesion molecule L1 in axonal growth cones. J Neurosci 20: 3676-3686, 2000.
Calcium signaling in growth cone induced by diffusible guidance molecules
Growth cone navigation is an essential process in establishing neural connections in the developing nervous system. Nerve growth cones are guided to specific targets by substrate-bound or diffusible guidance molecules. Most known guidance molecules have bi-functional roles, serving either as attractants or repellents, depending on the developmental stage or the specific classes of neurons they encounter. Using the nerve growth cone turning assay, combined with whole-cell recordings and Ca2+ imaging of Xenopus spinal neuronal growth cones, our studies seek to understand the molecular mechanisms underlying the bi-functional roles of these guidance molecules. Our studies have found that Ca2+ influx through voltage-dependent Ca2+ channels and Ca2+-induced Ca2+ release from internal Ca2+ stores tightly regulate intracellular Ca2+ levels which, in turn, mediate the polarity of growth cone turning in response to the diffusible guidance molecules netrin-1 and Sema3A. However, different classes of Ca2+ channels respond to each of these distinct guidance signals, suggesting that coordinated regulation of guidance molecules and Ca2+ channels account for specific neuronal guidance during development.
- Nishiyama M, Hoshino A, Tsai L, Henley JR, Goshima Y, Tessier-Lavigne M, Poo MM, Hong K. Cyclic AMP/GMP-dependent modulation of Ca2+ channels sets the polarity of nerve growth-cone turning. Nature. 2003 Jun 26;423(6943):990-5.
- Hong K, Nishiyama M, Henley J, Tessier-Lavigne M, Poo M. Calcium signalling in the guidance of nerve growth by netrin-1. Nature. 2000 Jan 6;403(6765):93-8.
- Hong K, Hinck L, Nishiyama M, Poo MM, Tessier-Lavigne M, Stein E. A ligand-gated association between cytoplasmic domains of UNC5 and DCC family receptors converts netrin-induced growth cone attraction to repulsion. Cell. 1999 Jun 25;97(7):927-41.
Multiple modulations of neuronal functions by brain-derived
neurotrophic factor (BDNF)
Some genes and chromosomal domains have been associated with risks for the illness, although no single gene or genes have been defined as causal for this complicated brain dysfunction. The gene for brain-derived neurotrophic factor (BDNF) has been suggested to be implicated in the risk for schizophrenia. Individual human has one or more different sequential variations (alleles) that are determined in part by past mutations and their permanent incorporation in the genome. This is also the case for BDNF gene. One frequent and non-conservative polymorphism for BDNF is a single nucleotide polymorphism (SNP) found in its pro-domain whose biological functional role remains to be fully understood. In the rodent brain, BDNF facilitate long-term potentiation in hippocampus and cortex, by enhancing synaptic transmission and vesicle docking. Therefore, we have looked for subtle cognitive and physiological differences between persons with different BDNF alleles and the underlying molecular mechanisms using several measures. Here I present our recent studies which demonstrate that different polymorphisms in the BDNF gene alter the distribution of its product, activity-dependent secretion manner, biological activity to decide neuronal survival or death, electrophysiological property to elicit synaptic transmission, and cognitive performance on tests of human learning and memory using an array of different tools. This integrative approach is of the most importance as it suggests that a polymorphism in neuronal genes modulates cognitive performance and clinical expression of brain dysfunction in human. In this lecture, I also talk about a novel mechanism of BDNF signaling through its receptor TrkB on cell surface. One idea for specific signal-function coupling is that different signaling pathways may be transduced in different subcellular compartments. More specifically, it has been proposed that cholesterol/sphingolipid-rich micro-domains called lipid rafts make a specialized signaling platform in the plasma membrane, and therefore can transduce signals different from those in the non-raft membrane. Here we show that BDNF rapidly recruits full-length TrkB (TrkB-FL) receptor into cholesterol-rich lipid rafts from non-raft regions of neuronal plasma membranes. Translocation of TrkB-FL was blocked by Trk inhibitors, suggesting a role of TrkB tyrosine kinase in the translocation. Disruption of lipid rafts by depleting cholesterol from cell surface blocked the ligand-induced translocation. Moreover, disruption of lipid rafts prevented potentiating effects of BDNF on transmitter release in cultured neurons and synaptic response to tetanus in hippocampal slices. In contrast, lipid rafts are not required for BDNF regulation of neuronal survival. Therefore, ligand-induced TrkB translocation into lipid rafts may represent a signaling mechanism selective for synaptic modulation by BDNF in the CNS.
- "BDNF-induced Recruitment of TrkB Receptor into Neuronal Lipid Rafts:
Roles in Synaptic Modulation", S. Suzuki, T. Numakawa, K. Shimazu,
H. Koshimizu, L. Mei, B. Lu, and M. Kojima, The Journal of Cell
Biology, vol.167, 1205-1215, 2004
- "Variant brain-derived neurotrophic factor (BDNF) (Met66) alters the
intracellular trafficking and activity-dependent secretion of
wild-type BDNF in neurosecretory cells and cortical neurons", Chen ZY,
Patel PD, Sant G, Meng CX, Teng KK, Hempstead BL, Lee FS, The Journal
of Neuroscience, vol.24, 4401-4411, 2004
"Genes and the parsing of cognitive processes", Goldberg TE,
Weinberger DR. Trends. Cogn. Sci., vol.8, 325-335, 2004
- "Lipid Rafts Mediate Chemotropic Guidance of Nerve Growth Cones",
C. Guirland, S. Suzuki, M. Kojima, B. Lu, and J. Q. Zheng, Neuron,
vol.42, 51-62, 2004
- "The BDNF val66met polymorphism affects activity-dependent secretion
of BDNF and human memory and hippocampal function", M. F. Egan,
M. Kojima, J. H. Callicott, T. E. Goldberg, B. S. Kolachana,
A. Bertolino, E. Zaitsev, B. Gold, D. Goldman, M. Dean, B. Lu, and
D. R Weinberger, Cell, vol.112, 257-269, 2003
- "Lipid rafts and the control of neurotrophic factor signaling in the
nervous system: variations on a theme", Paratcha, G., and
C.F. Ibanez., Curr. Opin. Neurobiol., vol.12, 542-549, 2002
- "Biological characterization and optical imaging of brain-derived
neurotrophic factor-green fluorescent protein suggest an
activity-dependent local release of brain-derived neurotrophic factor
in neurites of cultured hippocampal neurons", M. Kojima, N. Takei,
T. Numakawa, Y. Ishikawa, S. Suzuki, T. Matsumoto, R. Katoh-Semba,
H. Nawa, and H. Hatanaka, Journal of Neuroscience Research, vol.64,
- "Lipid rafts and signal transduction", Simons, K., and
D. Toomre. Nat. Rev. Mol. Cell Biol. vol.1, 31-39, 2000