神経細胞(ニューロン) の構造naibunpi/oka/komabalecture-1.pdf2009 sfn chicago, poster...
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• "Neuroscience“ Editors: Dale Purves et al., Sinauer, (ISBN: 978-0-87893-695-3)Hardcover 5th edition (2012)
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•20121212 (ISBN9787878-88-4-44-4444--2747474-444-211959595-55-9)
• http://ssl.ohmsha.co.jp/cgi-bin/menu.cgi?&ISBN=978-4-274-21195-9
•2015
•2006
•
• From Neuron To Brain. 4th Ed. Nicholls, J.G., et al., Sinauer, 2001 (ISBN 0-87893-439-1)
• Ion Channels of Excitable Membranes. 3rd
Ed. Hille, B. Sinauer, Associates, 2001 (ISBN 0-87893-321-2)
,
•
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••• e-mail: [email protected]• http://www.bs.s.u-tokyo.ac.jp/~naibunpi/
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Society for Neuroscience
SfN 2011, DC
Neuroscience 2010 by the numbers:31,975 attendees
Society for Neuroscience 2009399thth Annual Meeting, Chicago (29,980 attendees)
2009 SfN Chicago, Poster Sessions
McCormick Place, Chicago Convention Center
Special Lecture, President T. Carew
Lake Michigan
McCormick Place
Society for Neuroscience 2012422ndnd Annual Meeting, New Orleans (28,505 attendees)
2012 SfN New Orleans
Drs. L. Jan & Y.N. Jan Poster Sessions Eric Kandel
2008.10.09
GFPFP-P-GnRHH medakaTransgenic GFP-GnRH1 neurons Transgenic GFP-GnRH3 neurons
Medaka
•Genome database•Transparent•Spawn daily
Tg gnrh1
Tg gnrh2
Tg gnrh3
Tg gnrh2
Brain
Remove tectum
Electrode
Transgenic GFP-GnRH2 neurons
Whole brain in vitroOkubo et al. (2006)
Kanda et al. (2010)
Pituitary
POA
Spawn da
i GFWhole brain
G RFP
ailydan in vitro
RH2RHo prep
GFP
WT F0 ( )
F1
MedakaIn vivooo Caaa2+++++
–– –
Inverse pericamCRISPR-Cas9 / TALEN
GFP / -
1
GFP / GFPGFP / --/ -
GFP / -
F0
F1
F2
F2
Kiss1/kiss2
SV40 polyAPcmcl2DsRed
SV40polyA
F2F2
Daily cyclicity in pituitary lh gene expression
lhb mRNA
POAA-A-GGGG RH1 neuron activity y and d lhhh gene POAAA GGGG RH1 neuroRexpression show
on activityy ndandeurow w daily cyclicity
Daily cyclicity in AP firing activity of GnRH1 neurons
Karigo et al., 2012
Spontaneous action potential(AP) firing13:00
16:00
may be related to preovulatory GnRH/LH surge may be related to preovulator DL (spawning) cyclicity
IP
LH FSH Ca2+
LHβ-IP
FSHβ-IP
50μm
50μm
LH
Ca
FSHCa
Baseline peak 20-80%
Cells from 6 fish
173 147
BBas peak 20 80selineseelinese
[Ca2+]i GnRH
LH
FSH
http://www.nobelprize.org/nobel_prizes/medicine/laureates/2013/
(10/04/2004)
http://nobelprize.org/
200002000
(10/04/2004)200404200004
200002000
20000020000
199191199991http://sites.sinauer.com/neuroscience5e/animations04.01.html
2004
1.
2.
3.
4.
10/09 00:37
Fig 1. The dividing wall between the cell and the outside world – including other cells – is far from being an impervious shell. On the contrary, it is perforated by various channels. Many of these are specially adapted to one specific ion or molecule and do not permit any other type to pass. Here to the left we see a water channel and to the right an ion channel.
Fig 2. Peter Agre’s experiment with cells containing or lacking aquaporin. The aquaporin is necessary for making the cell absorb water and swell.
Brain•••
2004 Sinauer Associates, Inc.
(Kole, 2011)
2004 Sinauer Associates, Inc.
(A) – (D)
(G), (H)
(E), (F)
(I)(I)(J) ( ) ( )(K)
(L)
(M) (N) (O)
(P)
2004 Sinauer Associates, Inc.
A/D D/A
1.5mm
Patch pipettes
m
in vitro
ON
Tel.
Cut
OB
rostral
caudal
Cut
OT
••
••
(mV
)(n
A)
2004 Sinauer Associates, Inc.
-90
-80
-70
-60
-50
-40
-30
-20
-10
0
(mV
)(n
A)
A B
E
C
D
V=Vmax 1- exp(-t/RC)RC=
nA
mV
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nA
0mV
-65mV
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G
+/- 0.05 nA +/- 0.10 nA +/- 0.15 nA
F
Vm
ax/V
V=Vmax 1- exp(-t/ )
2004 Sinauer Associates, Inc.
G
E=0 E=EK
K+K+
A- A-
K+ K+
A- A-
---- +
+
++
K+
1
221 ][
][lnKK
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K+
K+
Nernst
K+ =
K+K+ K+
→
K+
58mV
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SEsR (8.315joule/deg mol)T 20℃ 293K)F (96,480coul/mol)Goldman-Hodgkin-Kat
oCliNaik
iCloNaok
oCliNaik
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ClPNaPKPClPNaPKP
ClPNaPKPClPNaPKP
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][][][][][][log58
][][][][][][ln
10
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1
221 ][
][log303.2][][ln
SS
FZRT
SS
FZRTEEEs
ss
K+ Na+
(mV)
-75+55(-60;Clin=52)
2004 Sinauer Associates, Inc.
(mV
)(m
V)
(mV
)(m
V)
(mV
)
Na+
58mV
(mV
)(m
V)
K+
58mV
2004 Sinauer Associates, Inc.
20 mTN-GnRH neurons
100 m
m
GnRH-3
A B
Neher & Sakmann, 1991
1~2pA
0pA
ABNa+
Na+
Na+ Na+
Na+
Whole-cellI
I=i1+i2+ in
i2 i1i3
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i5
in
in-1
Na+
3 Na+
10 Na+
Na+
-60mV
0mV
1pA
1pA
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100nA~
i
On-cell
i
Na+ K+
(mV
)
(mV
)
(mV
)
Na+
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Na+
(nA
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a+
(nA
)
K+
(nA
)K
+
(mA
/cm
2 )
Na+ K+
0
0.5
1
0
0.5
1
2004 Sinauer Associates, Inc.
Hodgkin, A. & Huxley, A.1963
(Eccles, J.C. )
2004 Sinauer Associates, Inc.
(Vm) Vm (Vm)
(mV
)(m
A/c
m2 )
65mV
65mV
(mA
/cm
2 )(m
V)
2004 Sinauer Associates, Inc.
(mV
)(m
A/c
m2 )
(mV
)(m
A/c
m2 ) TEA
A
B C
D
0
0.5
1
(mV)
TTX Na
(mA
/cm
2 )
(mV)
Na K
-40
-30
-20
-10
0
10
20
-60 -30 0 25 50 60
Na INa=gNa*(Vm-ENa )
g_Na*(V_m-E_Na )
Na+
(Vm) -60 -30 0 25 50 600 0.15 0.75 1 1 1
(Vm-ENa ) -110 -80 -50 -25 0 10gNa *(Vm-ENa ) 0 -12 -37.5 -25 0 10
0
20
40
60
80
100
120
140
-60 -30 0 25 50 60
K Ik=gK*(Vm-EK )g_K*(V_m-E_K )
K+
(Vm) -60 -30 0 25 50 600 0.15 0.75 1 1 1
(Vm-EK) 0 30 60 85 110 120gK*(Vm-EK ) 0 4.5 45 85 110 120
(mV
)(m
A/c
m2 )
Na+
(mS/
cm
2 )K
+(m
S/ c
m2 )
2004 Sinauer Associates, Inc.(m
V)
(mSi
emen
s/cm
2 )
(mV
)
Na+ K + 7, 15, 90 mV
Na+
Na+
K+
K+
(mSi
emen
s/cm
2 )
mV
2004 Sinauer Associates, Inc.
Na+
2m: Na+
h: Na+
n: K+
(mV
)(m
V)
(mm)
(mm)
(mV
)(m
V)
→→
=1/e
2004 Sinauer Associates, Inc.
Na1
Na3
2
Na K45
2004 Sinauer Associates, Inc.
2004 Sinauer Associates, Inc.
Vx=V0 exp(-x/ )
=
Vx:
V0: length constant,
rm: ri:
ro:
Rm:
Ri
rm=Rm/2 ari=Ri/ a2
=
d=2a d: a:
cRNA
cRNA
(A) (B)(C) mRNA
K+
K+
(mV
)(
A)
2004 Sinauer Associates, Inc.
Hydropathic index→ hydrophobicity=
Na+ Channel K+ ChannelNa+
K+
S
Na+
2004 Sinauer Associates, Inc.
ShB
R: ArgK: Lys
(E: Glu)
2004 Sinauer Associates, Inc.
2004 Sinauer Associates, Inc.
KThe Structure of the Potassium Channel:
Molecular Basis of K+
Conduction and SelectivityDeclan A. Doyle, Joa˜ o Morais Cabral,
Richard A. Pfuetzner, Anling Kuo, Jacqueline M. Gulbis, Steven L. Cohen, Brian T. Chait,
Roderick MacKinnon* SCIENCE VOL. 280 3 APRIL 1998
Ion conduction pore is conserved among
potassium channelsZhe Lu, Angela M. Klem & Yajamana RamuNATURE |VOL 413 | 25 OCTOBER 2001
Bacterial K channelKcsA
2004 Sinauer Associates, Inc.
Na
The voltage-sensitive sodium channel is a bell-shaped molecule with several cavitiesChikara Sato*, Yutaka Ueno*, Kiyoshi Asai*, Katsutoshi Takahashi², Masahiko Sato³, Andreas Engelß & Yoshinori FujiyoshiNature 409:1047-1051(2001)
2004 Sinauer Associates, Inc.
2004 Sinauer Associates, Inc.