adventurous motility - rice universitybioewhit/courses/bioe592/mat/brownell/synapse2003...
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Synaptic Transmission in Hair CellsW.E. Brownell
Sensory Neuroengineering, Rice8 October 2003
Hypothesis:OHC electromotility evolved from
hair cell synaptic mechanisms.
Edge of a Myxococcus xanthus colony - individual bacteria showing adventurous gliding motility, time lapse 600x speed (Kaiser lab website - Stanford).
Adventurous Motility
Outer membrane ripples on motile cells: Coincidence or functional roles?
OHC -Dieler et al. 1991
Oscillatoria -Adams et al. 1999
Flexibacter BH3 -Dickson et al. 1980
Trilaminate Walls
Oscillatoria –Adams et al. 1999
OHC
Neural membrane curvature Mechano-Electrical Transduction
Neurotransmission
Hair Cells Have Two Functions
Hair Cell Neurotransmission
Hudspeth, 1983Compton’s Interactive Encyclopedia, 1997
The output is neural
Frequencyincrease
The organ of Corti SensitiveSynapse
• Continuous release of neurotransmitter
• Rate of release modulated by < mV changes in membrane potential
Patterns - hair cells & photoreceptors
connectingcilium
kinocilium
Light dependent transmitter release
Dark – more release Light – less release
Schaeffer & Raveola, 1978
Phototransduction - dark current
The dark current depolarizes the membrane potential resulting in maximal neurotransmitter release. Light blocks the depolarizing current and decreases neurotransmitter release.
Visualizing the silent current
Geisler, 1974
Inner hair cell afferent synapse
Siegel & Brownell - 1986
Ribbon synapses
Lenzi & von Gersdorff, 2001
Ampullary organ of the North American Catfish
Mullinger, 1964
3-D reconstruction of frog hair cell synaptic ribbon
Lenzi et al. 1999
Recording from the synapse
Glowatzki & Fuchs, 2002
Interval between neurotransmitterrelease is Poisson
Glowatzki & Fuchs, 2002
Temporal precision
Kiang et al., 1965
Phase locking
Brownell, 1975
Intensity - invariance
Anderson, 1971
Specialized CNS synapses
Rowland et al., 2000
Preserve temporal coding
Protein-Protein Interactions in the Active Zone Matrix
Martin, 2002
Proteins bring membranes together
Weber et al., 1998
Vesicle fusion
Torri-Tarelli et al., 1985
Membrane fusion
Markin & Albanesi, 2002
(A) Parameters of the stalk.(B) Hemifusion,
- initial stage.(C) Hemifusion,
- transmonolayers contact.(D) Complete fusion
- fusion pore.
The fusion pore
attached to cellular membrane
Farrell & Cox, 2002
Geometry
xy
z
0 30-30-10
10
0
x (nm)
z (n
m)
0 30-30-10
10
0
x (nm)
cytosol cytosol
vesicle
extracellular
xy
z xy
z
0 30-30-10
10
0
x (nm)
z (n
m)
0 30-30-10
10
0
x (nm)
cytosol cytosol
vesicle
extracellular
Model
Ener
gy
Pore Reaction Coordinate
formation
fluctuation
expansion
50 k
T
Where do we get the energyto bend the membranes?
Phospholipids:the forgotten molecules
Don’t forget water
Membrane self assembly
Marrink, Lindahl, Edholm & Mark, 2001
+25 ns
Surface tension
attr
actio
n←
Ener
gy →
repu
lsio
n
Intermolecular distance
warmer
cooler
(
µ,
∂γ=−∂G
)TA
the energy required to increase the surface area of a liquid by a unit amount
Electrical potential changes γ:Lippmann mercury voltmeter
G. Lippmann, Ann. Phys. 149 (1873) DC Grahame (1947)
Voltage dependent membrane tension
Petrov & Sachs, 2002
Includes a differential change in surface tension at the two membrane interfaces
HEK electromotilitymeasured under voltage clamp with AFM
Zhang et al., 2001Mosbacher et al., 1998
V(+)
Voltage dependent membrane motion
Zhang et al., 2001
Voltage dependentpressure changes in squid axon
Terakawa, 1984
Electrical potential changes γ:Lippmann mercury voltmeter
G. Lippmann, Ann. Phys. 149 (1873) DC Grahame (1947)
Lippmann equation
Gabriel LippmannNobel Prize, physics 1908
A Gibbs adsorption equation for a polarizable interface,
∂γ=− ∂ −Γ∂µ−σ ∂oAS T E
:
o
-2 )
-2surface charge (Cm )-1: surface tension (Nm )
: electrical potential difference (V): chemical potential (V): temperature ( K ):surface concentration one component (moles m
:int erfacial entr
σ
γ
µ
Γ
o
A
E
T
S 2-1opy per unit area (JK m )−
contains the observed relation between surface charge and the ratio of the change in surface tension to the change in electrical potential,
( , )
ο
µ
∂γσ −∂
=TE
( , )
ο
µ
∂γσ −∂
=TE
Integrate the Lippmann under these boundary conditions:
εε κo=Ci
2j j2
j o
n z
RT
εε
= ∑2κ F1.
2.
p
dd oi i i i
ddm mi i i i
when
o e e e e
e e e ep
when and when
and
(V ) (V )
- q ) (V ) + q ) (V )
γ =γ γ =γ
γ =γ γ =γ
οο
οο
σ σ σ σ
σ σ σ σ
m
At voltage
Upon polarization to voltage V
= - = -
= -( = -(
oV
Voltage Dependent Tension
: j j
o w
Faraday's constant; n : concentration of species, j; z valency of species, j; R: gas constant;: permittivity of free space; : dielectric constant of water; : capacitance of double layer
ε ε
F
C
22 22
1 1
o om e m e ei i iV
e ie oo i
(C )T B B ( V ) C B B V
B B
≈ + ∆ + − σ + σ ∆
=± =±κ εε κ εε
Assume:Physiological medium 0.14 M External and internal surface charge -0.025 and -0.015 C/m2
Charging occurs by ions adsorbing onto or desorbing from∆V = 100 mV
Energy ≅ Pore Area * TV1000-5000 nm2 * 46µN/m10-50 kT
Tension is a linear function of voltage under physiological conditions (∆V< 100 mV)
: ; mvoltage dependent tension C membrane capacitance:VT
Flexoelectricity:coupling of membrane curvature of with the electric field
characterized in biological membranesby Petrov
Todorov, 1993
The flexoelectric effect
pillar
Plasma membrane
Spectrin
∆c
Vm1 Vm2
+≈
L
D
W
Cb
fffcUεεε0
D Cb
o w L
c membrane curvature; f flexoelectric coefficient (dipole); f flexoelectric coefficient (charge);: permittivity of free space; : dielectric constant of water; :dielectric constant of m: : :
ε ε ε embrane
Liquid Crystal Nature of BiomembranesProtein and lipid molecules comprising biomembranes
possess dipole moments
Dipoles contribute to the flexoelectric effect• curvature deformation changes membrane polarization
EP
PE
As c is increased, dipoles become more aligned increasing the polarization of the membrane
Direct flexoelectric effect
pore fluidcytosol
fused mem
brane
Ps Ps
-0.005 -0.001-0.003
4
3
5
2
Uf(
x 10
3 , µV
) 1
0Cb(nm-1)
Uf
pore fluidcytosol
fused mem
brane
Ps PsPs Ps
-0.005 -0.001-0.003
4
3
5
2
Uf(
x 10
3 , µV
) 1
0Cb(nm-1)
-0.005 -0.001-0.003
4
3
5
2
Uf(
x 10
3 , µV
) 1
0-0.005 -0.001-0.003
4
3
5
2
Uf(
x 10
3 , µV
) 1
0Cb(nm-1)
Uf
+≈
L
D
W
Cb
fffcUεεε0
The synaptic amplifier
Intrinsic tuning: - electrical
- membrane cycling
OHC electromotility – the other membrane based motor
Baylor:Amy Davidson, Ph.D.Brenda Farrell, Ph.D.
Olivier Lichtarge, M.D., Ph.D.Fred Pierera, Ph.D.Peter Saggau, Ph.D.
Buffalo:Fred Sachs, Ph.D.
Ken SnyderJohns Hopkins:
Aleksander S. Popel, Ph.D. Alexander A. Spector, Ph.D.
Rice:Bahman Anvari, Ph.D.
Robert M. Raphael, Ph.D.URLs: http://www.bcm.tmc.edu/oto/research/cochlea/
http://www.bioflexoelectricity.org
Collaborators
Hofmeister effect
Clarke & Lüpfert, 1999ClO4 > SCN > I > NO3 > Br > Cl > F > SO4
anion adsorption at membrane interface
Oliver et al., 2001I > Br > NO3 > Cl > HCO3 > F > SO4
Tension affectsvesicle recycling
Dai et al., 1997
Hyperpolarization & depolarization affect the charge on each interface
cytoplasmicextracellularmembrane
V
x=∞ x=-∞x=0 x=-δ
ψt
RestHyperpolarization Depolarization
V = ψ(-∞) - ψ(∞)= ψ(- ∞)
ψt = ψ(-δ)- ψ(0)Ψt : potential difference across the membrane V : transmembrane potential difference
V
ψt
x=0 x=-δ
V
ψt
x=0 x=-δ