Wen-Hung Twan.ppt [相容模式] -...

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2009/8/15 1 Hormones and reproduction in Hormones and reproduction in scleractinian corals scleractinian corals Wen Wen-Hung Twan Hung Twan 1,2 1,2 , , and Pei Pei-Jie Meng Jie Meng 1,3 1,3 1 National Museum of Marine Biology & Aquarium, Pingtung 944, Taiwan 2 Institute of Marine Biotechnology, National Dong Hwa University, Pingtung 944, Taiwan 3 Institute of Marine Biodiversity and Evolution, National Dong Hwa University, Pingtung 944, Taiwan Introduction Patterns of Sexual reproduction Brooder Broadcaster (broadcast spawner) Synchronous mass spawning Environmental cues Seawater temperatures Lunar phase

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Hormones and reproduction in Hormones and reproduction in scleractinian coralsscleractinian corals

WenWen--Hung TwanHung Twan1,21,2, , and PeiPei--Jie MengJie Meng1,31,3

1 National Museum of Marine Biology & Aquarium, Pingtung 944, Taiwan2 Institute of Marine Biotechnology, National Dong Hwa University,

Pingtung 944, Taiwan 3 Institute of Marine Biodiversity and Evolution, National Dong Hwa

University, Pingtung 944, Taiwan

Introduction

Patterns of Sexual reproductionBrooderBroadcaster (broadcast spawner)

Synchronous mass spawning

Environmental cuesSeawater temperaturespLunar phase

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Coral and Seawater CollectionCoral and Seawater Collection

Euphyllia glabrescens

Euphyllia ancoras

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Ready to spawning

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mesentery

gonad

mesentery

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Sex Steroids inSex Steroids in E. ancoraE. ancora

Testosterone and Estradiol-17ββ

Sex steroids have significant effects

OH OH

on gonadal development and gametogenesis

O

TestosteroneTestosterone

HO

EstradiolEstradiol--1717ββ

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Polyps were collected by scrapingPolyps were collected by scraping

Tissues were homogenizedTissues were homogenized

Characterization of Steroids in CoralCharacterization of Steroids in Coral

Tissues were homogenizedTissues were homogenized

SupernatantSupernatant

Alumina Gel ChromatographyAlumina Gel Chromatography

Extracted with diethyl etherExtracted with diethyl ether

rp-HPLC

Radioimmunoassay

g p yg p y

Alumina Gel Chromatography

Solvent Systemumn

Solvent System

TestosteroneTestosteroneor

Alu

min

a G

el C

olu

Coral extract

1st 4th3th2nd 5th 6th

EstradiolEstradiol--1717ββ

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The retention time of the testosterone and coral extract in the HPLC

30

10

20

30

oste

rone

(pg/

mg

tissu

e)

20

40

60

80

12

mv

11

7

[1] cortisone[2] cortisol[3] 11-ketotestosterone[4] 11-deoxycortisol[5] 20 βS

0 10 20 30 40 50 60 700

Test

o

0 10 20 30 40 50 60 70

0

201098654321

min

[6] androstenidione[7] deoxycorticosterone[8] testosterone[9] 17α, 20β -dihydroxypregnenone[10] estradiol[11] 17α-hydroxyandrostenone[12] progesterone.

Sex steroids were existed in E. ancora and E. glabrescens.

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The annual Profiles of Sex Steroids The annual Profiles of Sex Steroids in in E. ancoraE. ancora

20

30

40

50 a

b

ababab

ab

tero

ne (n

g/g

tissu

e) a

A

The annual changes of Sex steroid hormonesconcentrations in E. ancora.

Mar-98 Apr-98 May-98 Jul-98 Oct-98 Jan-99 Apr-99 May-990

10 b

Test

ost

500

600

700 a

ng/g

tiss

ue)

aB

spawning spawning

Mar Apr May Jul Oct Jan Apr May0

100

200

300

400

Estra

diol

-17β

(n

b

c bccbcbc

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30

40

50

60

70ne

(ng/g

tis

sue)

The changes of Sex steroid hormones concentrations in E.glabrescens during the spawning period

300

350

400

450

tiss

ue)

0

10

20

Apr May Jun Jul Aug Sep

Test

ost

eron

0

50

100

150

200

250

Apr May Jun Jul Aug Sep

Est

radio

l (ng/g

Conjugated SteroidsConjugated Steroidsj gj g

Conjugation (formation of hydrophilic molecules) is an important step in steroid catabolism

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Steroids and conjugated steroids measured in seawater collected during the spawning periods

During Mass SpawningSpawning

Seawater (pg/ml)

Estradiol

Sea SurfaceAround Coral

20 3 ± 3 4

5.8 ± 0.9 2.7 ± 0.7

5 9 ± 0 8

One dayafter

Mass Spawning

One dayprior to

Mass Spawning

Testosterone glucuronide

Testosterone

Estradiol glucuronide

Average value (mean ± SEM) in seawater (3 yr)

20.3 ± 3.4 5.9 ± 0.8

━ ━

━0.8 ± 0.2 ━

Steroids and conjugated steroids measured in coral and seawater collected during the E. ancora spawning periods

During Mass Spawning

Seawater (pg/ml)

Estradiol glucuronide

Estradiol

Sea SurfaceAround Coral

642.8 ± 25.4

20 3 ± 3 4

Coral(ng/g tissue)

228 8 ± 9 8

5.8 ± 0.9 2.7 ± 0.7

5 9 ± 0 8

Testosterone glucuronide

Testosterone

Estradiol glucuronide

Average value (mean ± SEM) in seawater (3 yr) and coral (3 colonies, 2yr)

20.3 ± 3.4228.8 ± 9.8

10.5 ± 2.0

42.7 ± 3.0

5.9 ± 0.8

━ ━

━0.8 ± 0.2

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Steroids and conjugated steroids measured in seawater (around coral) collected during the spawning periods

During Spawningg p g

Seawater (pg/ml)

Estradiol

E. glabrescensE. ancora

20 3 ± 3 4

5.8 ± 0.9 2.3 ± 0.3

Release larvaeRelease eggs

Testosterone glucuronide

Testosterone

Estradiol glucuronide

Average value (mean ± SEM) in seawater (3 yr)

20.3 ± 3.4 ━

━ ━

━0.8 ± 0.2

Thank you for your attentionThank you for your attention