Abstract
To investigate the regulatory role of the cyp19a1b aromatase gene in the sexual differentiation of largemouth bass (Micropterus salmoides, LMB), we obtained the full-length cDNA sequence of cyp19a1b using rapid amplification of cDNA ends technique. Tissue expression characteristics and feedback with 17-β-estradiol (E2) were determined using quantitative real-time PCR (qRT-PCR), while gonad development was assessed through histological section observations. The cDNA sequence of LMB cyp19a1b was found to be1950 base pairs (bp) in length, including a 5′ untranslated region of 145 bp, a 3′ untranslated region of 278 bp, and an open reading frame encoding a protein consisting of 1527 bp that encoded 508 amino acids. The qRT-PCR results indicated that cyp19a1b abundantly expressed in the brain, followed by the gonads, and its expression in the ovaries was significantly higher than that observed in the testes (P < 0.05). After feeding fish with E2 for 30 days, the expression of cyp19a1b in the pseudo-female gonads (XY-F) was significantly higher than that in males (XY-M) (P < 0.05), whereas expression did not differ significantly between XX-F and XY-F fish (P > 0.05). Although the expression of cyp19a1b in XY-F and XX-F fish was not significantly different after 60 days (P>0.05), both exhibited significantly higher levels than that of XY-M fish (P<0.05). Histological sections analysis showed the presence of oogonia in both XY-F and XX-F fish at 30 days, while spermatogonia were observed in XY-M fish. At 60 days, primary oocytes were abundantly observed in both XY-F and XX-F fish, while a few spermatogonia were visible in XY-M fish. At 90 days, the histological sections’ results showed that a large number of oocytes were visible in XY-F and XX-F fish. Additionally, the gonads of XY-M fish contained numerous spermatocytes. These results suggest that cyp19a1b plays a pivotal role in the development of ovaries and nervous system development in LMB.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bai JJ, Li SJ (2018) Development of largemouth bass (Micropterus salmoides) culture[M]. John Wiley & Sons, Ltd, 2018, 421–429
Breton TS, Dimaggio MA, Sower SA, Berlinsky DL (2015) Brain aromatase ( cyp19a1b ) and gonadotropin releasing hormone ( gnrh2 and gnrh3 ) expression during reproductive development and sex change in black sea bass ( centropristis striata ) Comp. Biochem Physiol A-Mol Integr Physiol 181:45–53. https://doi.org/10.1016/j.cbpa.2014.11.020
Brion F, Le Page Y, Piccini B, Tong S, Chung B (2012) Brain aromatase (cyp19a1b) is a highly sensitive gene to estrogens and xeno-estrogens. Endocr Disrupt 2012(2)
Callard GV (1992) Autocrine and paracrine role of steroids during spermatogenesis: studies in squalus acanthias and necturus maculosus. J Exp Zool 2(261). https://doi.org/10.1002/jez.1402610204
Callard GV, Tchoudakova AV, Kishida M, Wood E (2001) Differential tissue distribution, developmental programming, estrogen regulation and promoter characteristics of cyp19 genes in teleost fish. J Steroid Biochem Mol Biol 79(1-5):305–314. https://doi.org/10.1016/s0960-0760(01)00147-9
Cao W, Yi S, Yang N (2021) Molecular cloning and differential expression of cyp19a1 in tetraploid and diploid loach, misgurnus anguillicaudatus. J Huazhong Agric Univ 40(4). https://doi.org/10.13300/j.cnki.hnlkxb.2021.04.020
Cavaco JEB, van Baal J, van Dijk W, Hassing GAM, Goos HJT, Schulz RW (2001) Steroid hormones stimulate gonadotrophs in juvenile male African catfish (clarias gariepinus)1. Biol Reprod 64(5):1358–1365. https://doi.org/10.1095/biolreprod64.5.1358
Chen Y, Zhou P, Zhang Z, Xie F, Cai M, Wang Y (2015) Cloning and expression of cyp19a / b gene in large yellow croaker larimichthys crocea. J Jimei Univ (Nat Sci) 20(2):81–89. https://doi.org/10.19715/j.jmuzr.2015.2.001
Coumailleau P, Pellegrini E, Adrio F, Diotel N, Cano-Nicolau J, Nasri A, Vaillant C, Kah O (2015) Aromatase, estrogen receptors and brain development in fish and amphibians. Biochim Biophys Acta-Gene Regul Mech 1849(2):152–162. https://doi.org/10.1016/j.bbagrm.2014.07.002
Du J, Zhou J, Li S, Shao J, Jiang P, Dong C, Bai J (2021) A pcr-based method for genetic sex identification and evidence of the xx/xy sex determination system in largemouth bass (Micropterus salmoides l.). Aquaculture. 545:737220. https://doi.org/10.1016/j.aquaculture.2021.737220
Du J, Zhou J, Li S, Shao J, Jiang P, Lei C, Song H, Bai J, Han L (2022) Timing of early gonadal differentiation and effects of estradiol-17β treatments on the sex differentiation in largemouth bass (Micropterus salmoides). Fish Physiol Biochem 48(3):805–815. https://doi.org/10.1007/s10695-022-01074-4
El-Greisy ZA, El-Gamal AE (2012) Monosex production of tilapia, oreochromis niloticus using different doses of 17α-methyltestosterone with respect to the degree of sex stability after one year of treatment. Egypt J Aquat Res 38(1):59–66. https://doi.org/10.1016/j.ejar.2012.08.005
Garcia-Segura LM, Naftolin F, Hutchison JB, Azcoitia I, Chowen JA (1999) Role of astroglia in estrogen regulation of synaptic plasticity and brain repair. J Neurobiol 40(4):574–584. https://doi.org/10.1002/(SICI)1097-4695(19990915)40:4<574::AID-NEU12>3.0.CO;2-8
Gupta S, Moulik SR, Pal P, Majumder S, Das S, Guha P, Juin SK, Panigrahi AK, Mukherjee D (2017) Estrogen-regulated expression of cyp19a1a and cyp19a1b genes in swim-up fry of labeo rohita. Gen Comp Endocrinol 251:85–93. https://doi.org/10.1016/j.ygcen.2017.06.026
Hutchison JB, Beyer C, Hutchison RE, Wozniak A (1997) Sex differences in the regulation of embryonic brain aromatase. J Steroid Biochem 61(3):315–322. https://doi.org/10.1016/S0960-0760(97)80029-5
Karkola S, Höltje H, Wähälä K (2007) A three-dimensional model of cyp19 aromatase for structure-based drug design. J Steroid Biochem Mol Biol 105(1-5):63–70. https://doi.org/10.1016/j.jsbmb.2006.11.023
Katchalski-Katzir E, Shariv I, Eisenstein M, Friesem AA, Aflalo C, Vakser IA (1992) Molecular surface recognition: determination of geometric fit between proteins and their ligands by correlation techniques. Proc Natl Acad Sci U S A 89(6):2195–2199. https://doi.org/10.1073/pnas.89.6.2195
Kazeto Y, Ijiri S, Place AR, Zohar Y, Trant JM (2001) The 5′-flanking regions of cyp19a1 and cyp19a2 in zebrafish. Biochem Bioph Res Co 288(3):503–508. https://doi.org/10.1006/bbrc.2001.5796
Kwon JY, Kim J (2013) Differential expression of two distinct aromatase genes (cyp19a1a and cyp19a1b) during vitellogenesis and gestation in the viviparous black rockfish sebastes schlegelii. Anim Cells Syst 17(2):88–98. https://doi.org/10.1080/19768354.2013.773941
Leguen II, Carlsson C, Perdu-Durand E, Prunet P, Part P, Cravedi JP (2000) Xenobiotic and steroid biotransformation activities in rainbow trout gill epithelial cells in culture. Aquat Toxicol 48(2-3):165–176. https://doi.org/10.1016/s0166-445x(99)00043-0
Lephart ED (1996) A review of brain aromatase cytochrome p450. Brain Res Rev 22(1):1–26. https://doi.org/10.1016/0165-0173(96)00002-1
Li G, Liu X, Zhang Y, Bei J, Lin H (2004) Cloning and expression of two cytochrome p450 aromatase genes in red-spotted grouper epinephelus akaara. Acta Zool Sin 50(5):791–799. https://doi.org/10.3969/j.issn.1674-5507.2004.05.015
Li W, Li G, Luo C (2009) Molecular cloning and tissue-specific expression analysis of brain p450 arom in ricefield eel(monopterus albus). Fisheries Sci 28(8):458–461. https://doi.org/10.16378/j.cnki.1003-1111.2009.08.013
Li Y (2019) Molecular cloning and expression study of brain aromatase in carassius auratus. HNU. https://doi.org/10.27118/d.cnki.ghesu.2019.000188
Liu J (1993) Reproductive physiology of cultured fish in China. China Agriculture Press, 22–36
Maruska KP, Butler JM, Anselmo C, Tandukar G (2020) Distribution of aromatase in the brain of the african cichlid fish astatotilapia burtoni : aromatase expression, but not estro-gen receptors, varies with female reproductive-state. J Comp Neurol 528(15):2499–2522. https://doi.org/10.1002/cne.24908
Moon H, Lim H (2014) Synthesis and screening of small-molecule a-helix mimetic libraries targeting protein–protein interactions heejo moon1 and hyun-suk lim1,2. Curr Opin Chem Biol 24:38–47. https://doi.org/10.1016/j.cbpa.2014.10.023
Paixão RV, Silva GF, Caetano AR, Cintra LC, Varela ES, O'Sullivan FLA (2022) Phylogenomic and expression analysis of colossoma macropomum cyp19a1a and cyp19a1b and their non-classical role in tambaqui sex differentiation. Gene. 843:146795. https://doi.org/10.1016/j.gene.2022.146795
Patil JG, Gunasekera RM (2008) Tissue and sexually dimorphic expression of ovarian and brain aromatase mrna in the japanese medaka (oryzias latipes): implications for their preferential roles in ovarian and neural differentiation and development. Gen Comp Endocrinol 158(1):131–137. https://doi.org/10.1016/j.ygcen.2008.05.016
Pellegrini E, Diotel N, Vaillant-Capitaine C, Pérez Maria R, Gueguen M, Nasri A, Cano Nicolau J, Kah O (2016) Steroid modulation of neurogenesis: focus on radial glial cells in zebrafish. J Steroid Biochem Mol Biol 160:27–36. https://doi.org/10.1016/j.jsbmb.2015.06.011
Simpson ER, Mahendroo MS, Means GD, Kilgore MW, Hinshelwood MM, Graham-Lorence S, Amarneh B, Ito Y, Fisher CR, Michael MD (1994) Aromatase cytochrome p450, the enzyme responsible for estrogen biosynthesis. Endocr Rev 15(3):342. https://doi.org/10.1210/edrv-15-3-342
Sun J, Li G, Zhu C, Wu T, Deng S (2012) Molecular cloning and expression of cyp19a1b cdna in clarias fuscus. J Fish China 19(3):408–415. https://doi.org/10.3724/SP.J.1118.2012.00408
Tao W, Yuan J, Zhou L, Sun L, Sun Y, Yang S, Li M, Zeng S, Huang B, Wang D (2013) Characterization of gonadal transcriptomes from nile tilapia (oreochromis niloticus)reveals differentially expressed genes. PloS One 8(5):e63604. https://doi.org/10.1371/journal.pone.0063604
Tchoudakova A, Callard GV (1998) Identification of multiple cyp19 genes encoding different cytochrome p450 aromatase isozymes in brain and ovary. Endocrinology. 139(4):2179–2189. https://doi.org/10.1210/endo.139.4.5899
Tzchori I, Degani G, Hurvitz A, Moav B (2004) Cloning and developmental expression of the cytochrome p450 aromatase gene (cyp19) in the european eel (anguilla anguilla). Gen Comp Endocrinol 138(3):271–280. https://doi.org/10.1016/j.ygcen.2004.06.007
Valle LD, Ramina A, Vianello S, Belvedere P, Colombo L (2002) Cloning of two mrna variants of brain aromatase cytochrome p450 in rainbow trout (oncorhynchus mykiss walbaum). J Steroid Biochem Mol Biol 82(1):19–32. https://doi.org/10.1016/S0960-0760(02)00143-7
Wang D, Zhang G, Wei K, Gardner JPA, Yang R, Chen K, Zhou G (2015) Molecular cloning, characterization and expression analysis of brain cyp19a1b in channa argus. Fish Sci 45(6):9–15. https://doi.org/10.13721/j.cnki.dsyy.2015.06.002
Xing Y, Chen H, Li S, Guo X (2013) In vitro and in vivo investigation of a novel two-phase delivery system of 2-methoxyestradiol liposomes hydrogel. J Liposome Res 24(1):10–16. https://doi.org/10.3109/08982104.2013.822395
Xu P, Yu J, Li J, Xia D (2005) Molecular cloning of neural p450arom and its expression in pelteobagrus fulvidraco. J Fish China 29(5):591–598. https://doi.org/10.3321/j.issn:1000-0615.2005.05.002
Yamaguchi A, Tsunematsu T, Motojima Y, Toriyama K, Horinouchi A, Ishii Y, Murata H, Yoshikawa S, Nyuji M, Shimizu A (2022) Pituitary luteinizing hormone synthesis starts in aromatase (cyp19a1b)-positive cells expressing esr1 and esr2b at the onset of puberty in takifugu rubripes (fugu). Cell Tissue Res 389(2):259–287. https://doi.org/10.1007/s00441-022-03629-6
Yoshimoto FK, Guengerich FP (2014) Mechanism of the third oxidative step in the conversion of androgens to estrogens by cytochrome p450 19a1 steroid aromatase. J Am Chem Soc 136(42):15016–15025. https://doi.org/10.1021/ja508185d
Yu J, Wu T, Li J, Cao L, Xia D (2005) Cloning and sequence aanlysis of the cdna encoding p-450 aromatase from rice field eel. Acta Hydrobiol Sin 29(5):550–556. https://doi.org/10.1111/j.1440-1789.2005.00601.x
Zhou JH, Du JX, JP et al (2021) Effects of 17α-methyltestosterone on growth and sex differentiation in largemouth bass (Micropterus salmoides) [J]. J Fish Sci Chin 28(9):1109–1117
Availability of data and materials
All data and materials used to arrive at the conclusions of this paper are present in this manuscript.
Code availability
Not applicable.
Funding
This research was funded by Special-funds Project for Rural Revitalization Strategy of Guangdong Province (2022SPY00003); Science and Technology Planning Project of Guangzhou (202002030047); Guangdong Basic and Applied Basic Research Foundation (2020A1515010304); and Science and Technology Program of Guangzhou, China (2024B03J1300, 202201010211).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Ethical approval
This study was approved by the Ethics Committee of Animal Experiments of the Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences (CAFS), China.
Consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Zhang, D., Tian, T., Han, L. et al. Expression characteristics of the cyp19a1b aromatase gene and its response to 17β-estradiol treatment in largemouth bass (Micropterus salmoides). Fish Physiol Biochem 50, 575–588 (2024). https://doi.org/10.1007/s10695-023-01291-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10695-023-01291-5