Abstract
Diisononyl phthalate (DiNP) is a plasticizer used to improve plastic performance in a large variety of items which has been reported as an endocrine-disrupting chemical (EDC) in several organisms. The endocannabinoid system (ECS) is a cellular signaling system, whose functionality is tightly involved with reproductive function. The aim of the present study was the assessment of the effects of DiNP on the gonadal ECS and on the reproductive function of male gilthead sea bream Sparus aurata, an important marine aquacultured species in Europe, during the reproductive season. Fish were fed for 21 days with two diets contaminated with different nominal concentrations of DiNP (DiNP LOW at 15 µg DiNP kg−1 bw day−1 and DiNP HIGH at 1500 µg DiNP kg−1 bw day−1), based on the tolerable daily intake (TDI) ruled by the European Food Safety Authority for humans. The transcription of several genes related to the ECS was affected by the DiNP. Specifically, DiNP reduced the levels of endocannabinoids and endocannabinoid-like mediators, concomitant with the increase of fatty acid amide hydrolase (FAAH) activity. At the histological level, DiNP LOW induced the highest occurrence of individuals with regressed testes. Steroidogenesis was affected significantly, since plasma 11-ketotestosterone (11-KT), the main active androgen in fish, was significantly decreased by the DiNP HIGH treatment, while plasma 17β-estradiol (E2) levels were raised, associated with an increase of the gonadosomatic index (GSI). Additionally, the level of testosterone (T) was significantly increased in the DiNP LOW group, however, the same DiNP concentration reduced the levels of 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P). The production of sperm was in general not affected, since spermiation index, sperm density, survival and the duration of forward motility did not exhibit any changes compared to controls. However, computer-assisted sperm analysis (CASA) showed that DiNP reduced the percentage of motile cells. The results clearly suggest a negative effect of DiNP via the diet on the male endocrine system of gilthead sea bream during the reproductive season.
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Abbreviations
- DiNP:
-
Diisononyl phthalate
- ECS:
-
Endocannabinoid system
- EDC:
-
Endocrine disruptor chemical
- E2 :
-
17β-Estradiol
- T:
-
Testosterone
- 11-KT:
-
11-Ketotestosterone
- MIS:
-
Maturation-inducing steroid or 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P)
- AEA:
-
Anandamide
- 2-AG:
-
2-Arachidonoylglycerol
- PEA:
-
N-palmitoyl–ethanolamide
- OEA:
-
N-oleoyl-ethanolamide
- CB1/cnr1:
-
Endocannabinoid receptor type I
- CB2/cnr2:
-
Endocannabinoid receptor type II
- TRPV1:
-
Transient receptor potential cation channel subfamily V member 1
- PPAR:
-
Peroxisome proliferator-activated receptor
- NAPE-PLD:
-
N-acyl phosphatidylethanolamine phospholipase D
- FAAH:
-
Fatty acid amide hydrolase
- DAGLα:
-
Diacylglycerol lipase alpha
- COX-2 :
-
Cyclooxigenase-2
- ABDH:
-
α/β-Hydrolase
- CYT-PLA2:
-
Cytosolic phospholipase A2
- LEPR:
-
Leptin receptor
- LEPA:
-
Leptin A
- ER:
-
Estrogen receptor
- PR:
-
Progesterone receptor
- AR:
-
Androgen receptor
- LHR:
-
Luteinizing hormone receptor
- FSHR:
-
Follicle-stimulating hormone receptor
- GnRHR:
-
Gonadotropin-releasing hormone receptor
- 17β-HSD:
-
17β-Hydroxysteroid dehydrogenase
- 3β-HSD:
-
3β-Hydroxysteroid dehydrogenase
- VTGA:
-
Vitellogenin A
- ZP:
-
Zona pellucida proteins
- VCL:
-
Curvilinear velocity
- VSL:
-
Straight line velocity
- LIN:
-
Linearity
- VAP:
-
Average path velocity
- BW:
-
Body weight
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Acknowledgements
This work has received funding from Progetti di Rilevante Interesse Nazionale (PRIN) 2010–2011 (prot 2010W87LBJ) to OC and from the European Union’s Horizon 2020 research and innovation program under the TNA programme (Project ID AE030014) within AQUAEXCEL2020 project (652831) to IFP and OC for accessing to IATS-CSIC facilities. This output reflects only the author’s view and the European Union cannot be held responsible for any use that may be made of the information contained therein. The authors would like to thank the assistance of Marco Allarà for the enzymatic activity assay, Roberta Verde for assistance with the LC–MS/MS analyses and M. Ángeles González for technical assistance in PCR assays.
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Forner-Piquer, I., Mylonas, C.C., Fakriadis, I. et al. Effects of diisononyl phthalate (DiNP) on the endocannabinoid and reproductive systems of male gilthead sea bream (Sparus aurata) during the spawning season. Arch Toxicol 93, 727–741 (2019). https://doi.org/10.1007/s00204-018-2378-6
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DOI: https://doi.org/10.1007/s00204-018-2378-6
Keywords
- DiNP
- Endocannabinoids
- Gonads
- Gilthead sea bream