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Open Access 01-12-2008 | Research

Upregulated mRNA expression of desaturase and elongase, two enzymes involved in highly unsaturated fatty acids biosynthesis pathways during follicle maturation in zebrafish

Authors: Sairatul D Ishak, Sze-Huey Tan, Hou-Keat Khong, Annette Jaya-Ram, Yee-Ling Enyu, Meng-Kiat Kuah, Alexander Chong Shu-Chien

Published in: Reproductive Biology and Endocrinology | Issue 1/2008

Abstract

Background

Although unsaturated fatty acids such as eicosapentaenoic acid (EPA, C20:5n-3), docosahexaenoic acid (DHA, C22:6n-3) and arachidonic acid (ARA, C20:4n-6), collectively known as the highly unsaturated fatty acids (HUFA), play pivotal roles in vertebrate reproduction, very little is known about their synthesis in the ovary. The zebrafish (Danio rerio) display capability to synthesize all three HUFA via pathways involving desaturation and elongation of two precursors, the linoleic acid (LA, C18:2n-6) and linolenic acid (LNA, C18:3n-3). As a prerequisite to gain full understanding on the importance and regulation of ovarian HUFA synthesis, we described here the mRNA expression pattern of two enzymes; desaturase (fadsd6) and elongase (elovl5), involved in HUFA biosynthesis pathway, in different zebrafish ovarian follicle stages. Concurrently, the fatty acid profile of each follicle stage was also analyzed.

Methods

mRNA levels of fadsd6 and elovl5 in different ovarian follicle stages were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) assays. For analysis of the ovarian follicular fatty acid composition, gas chromatography was used.

Results

Our results have shown that desaturase displayed significant upregulation in expression during the oocyte maturation stage. Expression of elongase was significantly highest in pre-vitellogenic follicles, followed by maturation stage. Fatty acid composition analysis of different ovarian follicle stages also showed that ARA level was significantly highest in pre-vitellogenic and matured follicles. DHA level was highest in both late vitellogenic and maturation stage.

Conclusion

Collectively, our findings seem to suggest the existence of a HUFA synthesis system, which could be responsible for the synthesis of HUFA to promote oocyte maturation and possibly ovulation processes. The many advantages of zebrafish as model system to understand folliculogenesis will be useful platform to further elucidate the regulatory and mechanism aspects of ovarian HUFA synthesis.

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Wathes DC, Abayasekara DRE, Aitken RJ: Polyunsaturated fatty acids in male and female reproduction. Biol Reprod. 2007, 77 (2): 190-201. 10.1095/biolreprod.107.060558.CrossRefPubMed

Tahara D, Yano I: Maturation-related variations in prostaglandin and fatty acid content of ovary in the kuruma prawn (Marsupenaeus japonicus). Comp Biochem Physiol A. 2004, 137 (4): 631-637. 10.1016/j.cbpb.2003.12.005.CrossRef

Pickova J, Brännäs E, Andersson T: Importance of fatty acids in broodstock diets with emphasis on Arctic char (Salvelinus alpinus) eggs. Aquacult Int. 2007, 15 (3): 305-311. 10.1007/s10499-007-9084-3.CrossRef

Watanabe T, Vassallo-Agius R: Broodstock nutrition research on marine finfish in Japan. Aquaculture. 2003, 227: 1-4. 10.1016/S0044-8486(03)00494-0.CrossRef

Izquierdo MS, Fernandez-Palacios H, Tacon AGJ: Effect of broodstock nutrition on reproductive performance of fish. Aquaculture. 2001, 197 (1–4): 25-42. 10.1016/S0044-8486(01)00581-6.CrossRef

Patino R, Yoshizaki G, Bolamba D, Thomas P: Role of arachidonic acid and protein kinase C during maturation-inducing hormone-dependent meiotic resumption and ovulation in ovarian follicles of Atlantic Croaker. Biol Reprod. 2003, 68 (2): 516-523. 10.1095/biolreprod.102.009662.CrossRefPubMed

McEvoy TG, Coull GD, Broadbent PJ, Hutchinson JS, Speake BK: Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida. J Reprod Fertil. 2000, 118 (1): 163-170. 10.1530/reprod/118.1.163.CrossRefPubMed

Garrido S, Rosa R, Ben-Hamadou R, Cunha ME, Chicharo MA, Lingen van der CD: Effect of maternal fat reserves on the fatty acid composition of sardine (Sardina pilchardus) oocytes. Comp Biochem Physiol B. 2007, 148 (4): 398-409. 10.1016/j.cbpb.2007.07.008.CrossRefPubMed

Huynh MD, Kitts DD, Hu C, Trites AW: Comparison of fatty acid profiles of spawning and non-spawning Pacific herring, Clupea harengus pallasi. Comp Biochem Physiol B. 2007, 146 (4): 504-511. 10.1016/j.cbpb.2006.11.023.CrossRefPubMed

10.

Cejas JR, Almansa E, Villamandos JE, Badia P, Bolanos A, Lorenzo A: Lipid and fatty acid composition of ovaries from wild fish and ovaries and eggs from captive fish of white sea bream (Diplodus sargus). Aquaculture. 2003, 216 (1–4): 299-313. 10.1016/S0044-8486(02)00525-2.CrossRef

11.

Tocher DR, Agaba M, Hastings N, Bell JG, Dick JR, Teale AJ: Nutritional regulation of hepatocyte fatty acid desaturation and polyunsaturated fatty acid composition in zebrafish (Danio rerio) and tilapia (Oreochromis niloticus). Fish Physiol Biochem. 2002, 24 (4): 309-320. 10.1023/A:1015022406790.CrossRef

12.

Zheng X, Tocher DR, Dickson CA, Bell JG, Teale AJ: Effects of diets containing vegetable oil on expression of genes involved in highly unsaturated fatty acid biosynthesis in liver of Atlantic salmon (Salmo salar). Aquaculture. 2004, 236 (1–4): 467-483. 10.1016/j.aquaculture.2004.02.003.CrossRef

13.

Tocher DR: Metabolism and functions of lipids and fatty acids in teleost fish. Rev Fish Sci. 2003, 11 (2): 107-184. 10.1080/713610925.CrossRef

14.

Ling S, Kuah M-K, Tengku Muhammad TS, Kolkovski S, Shu-Chien AC: Effect of dietary HUFA on reproductive performance, tissue fatty acid profile and desaturase and elongase mRNAs in female swordtail Xiphophorus helleri. Aquaculture. 2006, 261 (1): 204-214. 10.1016/j.aquaculture.2006.06.045.CrossRef

15.

Jaya-Ram A, Kuah M-K, Lim P-S, Kolkovski S, Shu-Chien AC: Influence of dietary HUFA levels on reproductive performance, tissue fatty acid profile and desaturase and elongase mRNAs expression in female zebrafish Danio rerio. Aquaculture. 2008, 277 (3–4): 275-281. 10.1016/j.aquaculture.2008.02.027.CrossRef

16.

Wiegand MD: Composition, accumulation and utilization of yolk lipids in teleost fish. Rev Fish Biol Fish. 1996, 6 (3): 259-286. 10.1007/BF00122583.CrossRef

17.

Ge W: Intrafollicular paracrine communication in the zebrafish ovary: The state of the art of an emerging model for the study of vertebrate folliculogenesis. Mol Cell Endocrinol. 2005, 237 (1–2): 1-10. 10.1016/j.mce.2005.03.012.CrossRefPubMed

18.

Selman K, Petrino TR, Wallace RA: Experimental conditions for oocyte maturation in the zebrafish, Brachydanio rerio. J Exp Zool. 1994, 269 (6): 538-550. 10.1002/jez.1402690607.CrossRef

19.

Pang Y, Ge W: Gonadotropin and activin enhance maturational competence of oocytes in the zebrafish (Danio rerio). Biol Reprod. 2002, 66 (2): 259-265. 10.1095/biolreprod66.2.259.CrossRefPubMed

20.

Ziv T, Gattegno T, Chapovetsky V, Wolf H, Barnea E, Lubzens E, Admon A: Comparative proteomics of the developing fish (zebrafish and gilthead seabream) oocytes. Comp Biochem Physiol D. 2008, 3 (1): 12-35.

21.

Knoll-Gellida A, Andre M, Gattegno T, Forgue J, Admon A, Babin P: Molecular phenotype of zebrafish ovarian follicle by serial analysis of gene expression and proteomic profiling, and comparison with the transcriptomes of other animals. BMC Genomics. 2006, 7: 46-10.1186/1471-2164-7-46.PubMedCentralCrossRefPubMed

22.

Westerfield M: The Zebrafish Book: A Guide for the Laboratory Use of the Zebrafish (Danio rerio). 1994, Eugene OR: University of Oregon

23.

Selman K, Wallace RA, Sarka A, Xiaping Q: Stages of oocyte development in the zebrafish, Brachydanio rerio. J Morphol. 1993, 218 (2): 203-224. 10.1002/jmor.1052180209.CrossRef

24.

Bligh EG, Dyer WJ: A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959, 37: 911-917.CrossRefPubMed

25.

Ackman R: Fish lipids: part 1. Advances in Fish Science and Technology. Edited by: Connell J. 1980, Farnham, UK

26.

Retterstøl K, Tran TN, Haugen TB, Christophersen BO: Metabolism of very long chain polyunsaturated fatty acids in isolated rat germ cells. Lipids. 2001, 36 (6): 601-606. 10.1007/s11745-001-0763-z.CrossRefPubMed

27.

Ings JS, Kraak Van Der G: Characterization of the mRNA expression of StAR and steroidogenic enzymes in zebrafish ovarian follicles. Mol Reprod Dev. 2006, 73 (8): 943-954. 10.1002/mrd.20490.CrossRefPubMed

28.

Dai L, Ma W, Li J, Xu Y, Li W, Zhao Y, Deng F: Cloning and characterization of a novel oocyte-specific gene zorg in zebrafish. Theriogenology. 2008

29.

Essex-Fraser PA, Steele SL, Bernier NJ, Murray BW, Stevens ED, Wright PA: Expression of four glutamine synthetase genes in the early stages of development of rainbow trout (Oncorhynchus mykiss) in relationship to nitrogen excretion. J Biol Chem. 2005, 280 (21): 20268-20273. 10.1074/jbc.M412338200.CrossRefPubMed

30.

Hossain MS, Larsson A, Scherbak N, Olsson P-E, Orban L: Zebrafish androgen receptor: isolation, molecular, and biochemical characterization. Biol Reprod. 2008, 78 (2): 361-369. 10.1095/biolreprod.107.062018.CrossRefPubMed

31.

Billiau AD, Sefrioui H, Overbergh L, Rutgeerts O, Goebels J, Mathieu C, Waer M: Transforming growth factor β inhibits lymphokine activated killer cytotoxicity of bone marrow cells: implications of the graft-versus leukemia effect in irradiation allogeneic bone marrow chimeras. Transplantion. 2001, 71: 292-299. 10.1097/00007890-200101270-00022.CrossRef

32.

Ramachandra RK, Lankford SE, Weber GM, Rexroad CE, Yao J: Identification of OORP-T, a novel oocyte-specific gene encoding a protein with a conserved oxysterol binding protein domain in rainbow trout. Mol Reprod Dev. 2007, 74 (4): 502-511. 10.1002/mrd.20628.CrossRefPubMed

33.

Buschiazzo J, Bruzzone A, Alonso TS: Detailed lipid analysis of yolk platelets of amphibian (Bufo arenarum) oocytes. J Exp Zool Part A. 2003, 297 (2): 189-195. 10.1002/jez.a.10252.CrossRef

34.

Pati D, Habibi HR: Involvement of protein kinase C and arachidonic acid pathways in the gonadotropin-releasing hormone regulation of oocyte meiosis and follicular steroidogenesis in the goldfish ovary. Biol Repro. 2002, 66 (3): 813-822. 10.1095/biolreprod66.3.813.CrossRef

35.

Kraak Van Der G, Chang JP: Arachidonic acid stimulates steroidogenesis in goldfish preovulatory ovarian follicles. Gen Comp Endocrinol. 1990, 77 (2): 221-228. 10.1016/0016-6480(90)90306-7.CrossRefPubMed

36.

Sorbera LA, Asturiano JF, Carrillo M, Zanuy S: Effects of polyunsaturated fatty acids and prostaglandins on oocyte maturation in a marine teleost, the European Sea Bass (Dicentrarchus labrax). Biol Reprod. 2001, 64 (1): 382-389. 10.1095/biolreprod64.1.382.CrossRefPubMed

37.

Wang X, Walsh LP, Reinhart AJ, Stocco DM: The role of arachidonic acid in steroidogenesis and steroidogenic acute regulatory (StAR) gene and protein expression. J Biol Chem. 2000, 275 (26): 20204-20209. 10.1074/jbc.M003113200.CrossRefPubMed

38.

Wada M, DeLong CJ, Hong YH, Rieke CJ, Song I, Sidhu RS, Yuan C, Warnock M, Schmaier AH, Yokoyama C, et al: Enzymes and receptors of prostaglandin pathways with arachidonic acid-derived versus eicosapentaenoic acid-derived substrates and products. J Biol Chem. 2007, 282 (31): 22254-22266. 10.1074/jbc.M703169200.CrossRefPubMed

39.

Mercure F, Kraak Van Der G: Inhibition of gonadotropin-stimulated ovarian steroid production by polyunsaturated fatty acids in teleost fish. Lipids. 1995, 30 (6): 547-554. 10.1007/BF02537030.CrossRefPubMed

40.

Kennedy SR, Leaver MJ, Campbell PJ, Zheng X, Dick JR, Tocher DR: Influence of dietary oil content and conjugated linoleic acid (CLA) on lipid metabolism enzyme activities and gene expression in tissues of Atlantic Salmon (Salmo salar L.). Lipids. 2006, 41 (5): 423-436. 10.1007/s11745-006-5116-4.CrossRefPubMed

41.

Agaba M, Tocher DR, Dickson CA, Dick JR, Teale AJ: Zebrafish cDNA encoding multifunctional fatty acid elongase involved in production of eicosapentaenoic (20:5n-3) and docosahexanoiec (22:6n-3) acids. Mar Biotechnol. 2004, 6 (3): 251-261. 10.1007/s10126-003-0029-1.CrossRefPubMed

42.

Sprecher H, Luthria DL, Mohammed BS, Baykousheva SP: Reevaluation of the pathways for the biosynthesis of polyunsaturated fatty acids. J Lipid Res. 1995, 36 (12): 2471-2477.PubMed

43.

Hourvitz A, Gershon E, Hennebold JD, Elizur S, Maman E, Brendle C, Adashi EY, Dekel N: Ovulation-selective genes: the generation and characterization of an ovulatory-selective cDNA library. J Endocrinol. 2006, 188 (3): 531-548. 10.1677/joe.1.06231.CrossRefPubMed

44.

Kumadaki S, Matsuzaka T, Kato T, Yahagi N, Yamamoto T, Okada S, Kobayashi K, Takahashi A, Yatoh S, Suzuki H, et al: Mouse Elovl-6 promoter is an SREBP target. Biochem Biophys Res Commun. 2008, 368 (2): 261-266. 10.1016/j.bbrc.2008.01.075.CrossRefPubMed

45.

Seol HS, Sato K, Matsubara Y, Schneider WJ, Akiba Y: Modulation of sterol regulatory element binding protein-2 in response to rapid follicle development in chickens. Comp Biochem Physiol B. 2007, 147 (4): 698-703. 10.1016/j.cbpb.2007.04.012.CrossRefPubMed

46.

Steffensen KR, Robertson K, Gustafsson J, Andersen CY: Reduced fertility and inability of oocytes to resume meiosis in mice deficient of the Lxr genes. Mol Cell Endocrinol. 2006, 256 (1–2): 9-19. 10.1016/j.mce.2006.03.044.CrossRefPubMed

47.

Hastings N, Agaba M, Tocher DR, Leaver MJ, Dick JR, Sargent JR, Teale AJ: A vertebrate fatty acid desaturase with δ 5 and δ 6 activities. Proc Natl Acad Sci USA. 2001, 98 (25): 14304-14309. 10.1073/pnas.251516598.PubMedCentralCrossRefPubMed

48.

Moreau C, Froment P, Tosca L, Moreau V, Dupont J: Expression and regulation of the SCD2 desaturase in the rat ovary. Biol Reprod. 2006, 74 (1): 75-87. 10.1095/biolreprod.105.044545.CrossRefPubMed

49.

Cho HP, Nakamura M, Clarke SD: Cloning, expression, and fatty acid regulation of the human δ 5 desaturase. J Biol Chem. 1999, 274 (52): 37335-37339. 10.1074/jbc.274.52.37335.CrossRefPubMed

50.

Hastings N, Agaba MK, Tocher DR, Zheng X, Dickson CA, Dick JR, Teale AJ: Molecular cloning and functional characterization of fatty acyl desaturase and elongase cDNAs involved in the production of eicosapentaenoic and docosahexaenoic acids from α-linolenic acid in Atlantic Salmon (Salmo salar). Mar Biotechnol. 2004, 6 (5): 463-474. 10.1007/s10126-004-3002-8.CrossRefPubMed

51.

Tocher D, Zheng X, Schlechtriem C, Hastings N, Dick J, Teale A: Highly unsaturated fatty acid synthesis in marine fish: Cloning, functional characterization, and nutritional regulation of fatty acyl Δ6 desaturase of Atlantic cod (Gadus morhua L.). Lipids. 2006, 41 (11): 1003-1016. 10.1007/s11745-006-5051-4.CrossRefPubMed

52.

Agaba MK, Tocher DR, Zheng X, Dickson CA, Dick JR, Teale AJ: Cloning and functional characterisation of polyunsaturated fatty acid elongases of marine and freshwater teleost fish. Comp Biochem Physiol B. 2005, 142 (3): 342-352. 10.1016/j.cbpb.2005.08.005.CrossRefPubMed

53.

Clelland ES, Tan Q, Balofsky A, Lacivita R, Peng C: Inhibition of premature oocyte maturation: a role for bone morphogenetic protein 15 in zebrafish ovarian follicles. Endocrinology. 2007, 148 (11): 5451-5458. 10.1210/en.2007-0674.CrossRefPubMed

54.

Zhu Y, Rice CD, Pang Y, Pace M, Thomas P: Cloning, expression and characterization of a membrane progestin receptor and evidence it is an intermediary in meiotic maturation of fish oocytes. Proc Natl Acad Sci USA. 2003, 100 (5): 2231-2236. 10.1073/pnas.0336132100.PubMedCentralCrossRefPubMed

Title: Upregulated mRNA expression of desaturase and elongase, two enzymes involved in highly unsaturated fatty acids biosynthesis pathways during follicle maturation in zebrafish
Authors: Sairatul D Ishak
Sze-Huey Tan
Hou-Keat Khong
Annette Jaya-Ram
Yee-Ling Enyu
Meng-Kiat Kuah
Alexander Chong Shu-Chien
Publication date: 01-12-2008
Publisher: BioMed Central
Published in: Reproductive Biology and Endocrinology / Issue 1/2008
Electronic ISSN: 1477-7827
DOI: https://doi.org/10.1186/1477-7827-6-56

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Upregulated mRNA expression of desaturase and elongase, two enzymes involved in highly unsaturated fatty acids biosynthesis pathways during follicle maturation in zebrafish

Abstract

Background

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

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