and cell function Ca - 東京大学naibunpi/Oka/BiologicalSignaling...+( Ý ·"@Ca2+GOGqG[GyFþ (8®...

Ca2+ and cell function

2001 Sinauer Associates, Inc.

2002 Alberts et al.


2X10-3M (2mM)

10-7M(100nM)

10-4M(100 M)

Ca2+

Ca2+

Ca2+

http://sites.sinauer.com/neuroscience5e/animations07.02.html

Cells maintain a very low concentration of free Ca2+

in the cytosol

2002 Alberts et al.

Ca2+ Ca2+

Ca2+

Rhod-2Fura-2

Electroporation Calcium Green

0 20 40 60 80-0.2

0

0.2

0.4

0.6

0.8

1

Time (s)

DF/F0

#0#1#2

80400Time (s)

0

0.2

-60

-40

-20

0

20

40

60

Inverse e PPericamCa2+

Ca2+

Ca2+

LHβ /FSHβ

2kbInverse Pericam

PolyAsignal

LHβ-IP FSHβ-IP

IP

LH FSH Ca2+

LHβ-IP

FSHβ-IP

50μm

50μm

[Ca2+]i GnRH

LH FSH

[Ca2+]ii

CCD

Vh=-50mV

Exocytosis of GtH; blocked by EGTA/BAPTA in pipette


Na+ K+

(mV

)

(mV

)

(mV

)

Na+

K+

Na+

(nA

)N

a+

(nA

)

K+

(nA

)K

+

(mA

/cm

2 )

Na+ K+


Voltage-dependent Ca2+ channels

CsCl, TEA, EGTATTX 5mM Ca2+ A TTX 115mM Ba2+

B) -80mV -30mVC) -30mV

-80mV HVA, LVA

-30mV LVA

EH(-80)=HVA+LVAEH(-30)=HVA

HVALVA

HVALVA

-30mV LVAHVA


EH = -30mV

EC = -20mV

LVA(EH = -80mV)

LVA LVA

LVA(EH = -80mV→EC = -20mV)

EH

EH

EC

EC

Ca2+

tiny conductancetransient current

large conductancelong-lasting current


HVA HVA LVA

Tsien L P/Q N R T

α 1S 1C 1D 1F 1A 1B 1E 1G 1H 1I

CaV1.1CaV1.2CaV1.3CaV1.4CaV2.1CaV2.2CaV2.3CaV3.1CaV3.2CaV3.3

-30mV -20mV -70mV

( >500ms) ( 50~80ms) ( 20~50ms)

25pS 13pS 8pS

Cd2+>Ni2+ Cd2+>Ni2+ Ni2+>Cd2+

-CTX GVIA

-agatoxin

DHP

Ca2+

Hodgkin, A. & Huxley, A.1963

(Eccles, J.C. )

(stellate ganglion)


Ca2+

(mV)

CaA/cm2

(mV)


BAPTABAPTA

epsp EGTA

Ca2+, Ca2+ , Ca2+

Ca2+

Ca2+

EPSP

Ca2+

Ca2+

Ca2+

INa=gNa (Vm-ENa)

ICa=gCa (Vm-ECa)

Vm-ECa Tail current

gCa

INa IK

“Tail current”

-18mV+60mV

1ms 0.2ms

Ca2+ currents

Voltage clamp of presynaptic terminal

I2 vs V2

I1 vs V1

+TTX, TEA

+TTX, TEA

Ca2+

Ca2+

1 2

1:Ca2+

2:Ca2+

1+2=


gNa

gCa

-18mV +60mV

IK=gK (Vm-EK)

Vm=EK

Ca2+

off Ca2+ “tail current”

Channels and transporters for Ca2+

ThapsigarginX

IP3R


Ca2+ as a second messenger

triad

RyR

Phospholipase C- PI(4,5)P2

2002 Alberts et al.

IP3R RyR

GnRHCa2+

Ca2+

IP3 GnRH

Thapsigargin

Exocytosis of GtH; blocked by EGTA/BAPTA in pipette

Ca2+

: 10-7M[Ca2+]

Ca2+

Ca2+ Ca2+

10-7M~ 10-5M[Ca2+]

Thapsigargin

Thapsigargin

CRAC


(A)

Pituitary gland and Pituitary gland and neurosecretory cells

SONPVN

Hypophysiotropic hormones

Pituitary

neurosecretory y cells (neuroendocrine)

Hypophysiotropic hormone-producing neurons

Hypothalamus

Pituitary

Hypophysiotropic system (Brain-Pituitary interface)

Gonadotropic hormones

(Posterior pituitary hormones)

Gonadotropin-releasing hormone (GnRH) system

1 2 3 4 5 6 7 8 9 10mGnRH pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2

GnRH neurons

••

Protein and peptide hormones

Terminal Nerve GnRH System (sGnRH)

Midbrain GnRH System (cGnRH-II)

Hypophysiotrophic Preoptic GnRH System (sbGnRH)

Tel

TeO

ON

OB

Med

Pit

Cb

GnRH

Tel

Neuroendocrine

Neuromodulatory

Neuromodulatory

Gonadotropin-releasing hormone

pGlu-His -Trp-Ser-Tyr-Gly-Trp-Leu-Pro-Gly-NH2

pGlu-His -Trp-Ser-Hisr -Gly-Trp-Tyr-Pro-Gly-NH2c -II

pGlu-His -Trp-Ser-Tyr-Gly-Leu-Ser-Pro-Gly-NH2seabream

GnRH3

GnRH2

GnRH1

Three paralogous genes for GnRH--Synteny analysisisis-

Okubo & Nagahama, 2007

Ancestral vertebrates

Ancestral tetrapods

Medaka

Dwarf Gourami

Dwarf gourami

Ancestral teleosts

Medaka

Dwarf Gourami

Hypophysiotropic hormone-producing neurons

Hypothalamus

Pituitary

Hypophysiotropic system (Brain-Pituitary interface)

Gonadotropic hormones

(Posterior pituitary hormones)

Gonadotropin-releasing hormone (GnRH) system

1 2 3 4 5 6 7 8 9 10mGnRH pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2

GnRH neurons

1987 Kodansha Ltd.

Luteinizing hormone (LH)Follicle-stimulating hormone (FSH)Thyroid-stimulating hormone (TSH)

Growth hormone (GH)Prolactin (PRL)

Adrenocorticotropic hormone (ACTH)Melanocyte-stimulating hormone (MSH)

OxytocinVasopressin

Basophils (Glycoproteins)α

β

Acidophils

Chromatophobes

Pro-opiomelanocortin (POMC)

Protein and peptide hormones

Propressophysin Prooxyphysin

Neurophysin:

PC: proteolytic enzymesCLIP: corticotropin-like intermediate lobe peptide

LPH:

β-END: β


1987 Kodansha Ltd.

1987 Kodansha Ltd.

Weltzien et al., 2004

FSH

LH

IP

LH FSH Ca2+

LHβ-IP

FSHβ-IP

50μm

50μm

[Ca2+]i GnRH

LH FSH

[Ca2+]ii

CCD

Pituitary

Pituitary

GFPFP-P-labelled GFFPP belled alaGnRH1 neurons

Brain

gnrh1:EGFP; lhb:IP

Inverse e Pericam

Ca2+

Modified from Karigo, et al., 2014

Caa2+++ imaging using Double transgenic medakaka: Cagnrh1

aging usimaih1h1:EGFP

g usPP;

ing DsusPP;;; lhb

Doung Dhbhb:IP

Na1

Na3

2

Na K4

55


BioClassics5. /

Antidromic activation 1.2.

3.

VP

OX

A,B PVN

1,2 3,4

TNN-N-GnRHHTNNN-GGGnRHG HNN GGPacemaker potential)l)l)

Umatani et al., 2013 Kawai et al., 2013

Facilitatory modulation by GnRH3 peptide (Abe and Oka, 2000)

Saito et al., 2009

Inhibitory modulation by NPFF & RFRP

2 Steroid hormones

Aromatase

and cell function Ca - 東京大学naibunpi/Oka/BiologicalSignaling...+( Ý ·"@Ca2+GOGqG[GyFþ (8®...

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