Hormone Disrupting Chemicals 1993: Sharpe and Skakkebæk publicerer østrogen hypotesen
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1 Hormone Disrupting Chemicals 1993: Sharpe and Skakkebæk publicerer østrogen hypotesen We argue that the increasing incidence of reproductive abnormalities in the human male may be related to increased oestrogen exposure in utero, and identify mechanisms by which this exposure could occur. 1
2 Sperm cells 10 6 per ml Topari et al, 1996 Intersex in British Perch populations Jobling et al
3 Intersex Intersex Roach (Jobling et al, 2002) 3
4 Intersex Roach (Jobling et al, 2002) Xenoestrogens are chemically diverse 4
5 Xenoestrogen activity a result of 3D structure? A Model of an intracellular receptor COOH Inhibitory protien complex Hormone binding site Transcriptionactivating domain H 2 N Hinge region DNA-binding domain COOH H 2 N 5
6 The estrogen receptor domains Activating function 1 Nuclear Localisation signal Activating function 2 A/B C D E F DNA binding domain Hinge region Ligand binding domain Agonist binding domain Antagonist binding domain Homology between Human and Rainbow trout ER A/B: 20% C: 92% E: 60% Mathews et al Vitellogenin synthesis as a biomarker of estrogenic activity in water Gonadotropins 1 Estrogen 3 2 Liver Ovary 4 5 Vitellogenin 6
7 Possible sex homone disruption mechanisms: Estrogen Androgen Anti-Estrogen Anti-Androgen (Arukwe, 2001) Hormone disruption mechanisms Hormone synthesis Hormone secretion Hormone transport Receptor binding Receptor complex mediated transcription Hormone elimination -ve/+ve feedback loops 7
8 Steroid Pathways Made Simple Cholesterol P450 enzymes Xenoandrogens Progestogens Corticoids Androgens Estrogens Aromatase inhibitors Imposex in the dogwhelk may be due to aramatase inhibition 8
9 Androgen Effects mosquitofish caught below an american Pulp mill Androgens or aramatase inhibition?? Normal female Partly masculinised female Female with advanced masculinisation Normal male Parks et al, 2001 Papermill 9
10 Initial stages of female masculinisation Toft et al, 2004 Feminization is quantified by counting segments (Parks et al 2001) Reference females Polluted females Reference males 10
11 Downstream sites caused greater AR-binding inhibition than 1micromolar flutamide Parks et al, M Downstream sites caused similar AR-mediated gene expression as DHT Flutamide Parks et al,
12 The chemical induces AR migration into the nucleus Masculinization is not a result of corticosteroid receptor activity Dexamethasone potent glucocorticoidreceptor agonist Parks et al,
13 Brain and ovarian aromatase activity is higher in fish exposed to pulp mill effluent (Orlando et al., 2002) Ovary Brain Anti-Oestrogen effects Aromatase inhibition Receptor Crosstalk Between two intracellular receptors: Arylhydrocarbon receptor (AhR) & ER receptor AhR causes synthesis of P450 enzyme EROD ER induces production of vitellogenin in fish 13
14 Vitellogenin production in juvenile Coho salmon PCB Ah receptor Arukwe et al., 2001 Level of organization Effect Population Number of offspring Sex ratio Organism Organ Cell Time to maturation Sexual behaviour Colouration Testis size Gonopodial length Sperm count Molecule 14
15 The Guppy Mating System Highly promiscuous Continuous reproduction Viviparous, sperm-counts possible Female choice, male ornamentation display / courtship behaviour. Handicap models... Good gene models... Two different male mating strategies... Full display / Sneaky matings 15
16 Colouration index Gonopodial index The sigmoiddisplay Female Male 16
17 ε δ a ϕ b c ω frame (n) frame (n+1) 17
18 Sædtælling Sædtælling 18
19 Effects of antiandrogens Cl O Cl Cl O N Cl O Vinclozolin Cl p,p -DDE Cl Cl O O O Cl O OH N OH N HO N NO 2 Cl Cl O CF 3 M1 M2 Hydroxyflutamide Vinclozolins effects on Reproduction Effects on male fitness? Effects on female fitness? Effects during pregnancy? 19
20 Virgin + (30) male fitness traits (Sigmoid, sperm) Size of virgin females first clutch Guppy Reproduction Virgin (30) + Size of virgin females first clutch (pregnancy) + Size of virgin females first clutch Number sigmoid displays per 10 min Male sexual display 0,0 0,1 1,0 10,0 * Vinclozolin dose µg/mg dry feed Foetus Juvenile Adult ca. 30 days 20
21 Sædtal (millioner celler pr. han) 3,50 3,00 2,50 2,00 1,50 1,00 The effect of vinclozolin on sperm count ** *** 0,0 0,1 1,0 10,0 Vinclozolin dosis µg/mg tør foder Foetus Juvenile Adult ca. 30 days 20 Effect of vinclozolin on male fitness Number of juveniles per akvarium * ** 2 0,0 0,1 1,0 10,0 Vinclozolin dose µg/mg dry feed Foetus Juvenile Adult ca. 30 days 21
22 10 Male sperm count (millioner) Number of juveniles per male Guppy Reproduction Virgin + (30) male fitness traits (Sigmoid, sperm) Size of virgin females first clutch Virgin (30) + Size of virgin females first clutch (pregnancy) + Size of virgin females first clutch 22
23 Fitness?? Microsatellites and male competition experiments Use of genetic markers for assessing sex 23
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