Abstract
The molecular pathways involved in oviductal adenogenesis are highly conserved among vertebrates. In this work, we study the histomorphological changes and molecular pathways involved in Caiman latirostris oviductal adenogenesis and the effects of in ovo exposure to environmentally relevant doses of endosulfan (END) and atrazine (ATZ) on these processes. To this end, the histomorphological changes at epithelial and subepithelial compartments, the protein expressions of β-catenin and Wnt-7a, and the gene expression of metalloproteinases (MMPs) and its inhibitors (TIMPs) were evaluated as biomarkers of oviductal adenogenesis in prepubertal juvenile C. latirostris. Exposure to END altered adenogenesis-related epithelium characteristics and mRNA expression of MMP2, MMP9, and TIMP1. Exposure to ATZ increased the width of the subepithelial stroma with loosely arranged collagen fibers and increased β-catenin expression in buds (invaginated structures that precede glands). The results demonstrate that in ovo exposure to ATZ and END alters oviductal adenogenesis at tissue, cellular, and molecular levels. An altered oviductal adenogenesis could impair fertility, raising concern on the effects of pesticide pollution in wildlife and domestic animals.
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The datasets compiled and analyzed in the present study are available from the corresponding author on reasonable request.
Abbreviations
- ANOVA:
-
Analysis of variance
- ATZ:
-
Atrazine
- EDC:
-
Endocrine-disrupting compounds
- END:
-
Endosulfan
- CTNNB1:
-
β-Catenin encoding gene
- E2:
-
17β-Estradiol
- FoxA2:
-
Forkhead box protein A2
- IHC:
-
Immunohistochemistry
- MMP2:
-
Matrix metalloproteinase 2
- MMP7:
-
Matrix metalloproteinase 7
- MMP9:
-
Matrix metalloproteinase 9
- PAS:
-
Periodic acid Schiff
- PBS:
-
Phosphate-buffered saline
- TIMP1:
-
Tissue inhibitor of metalloproteinases 1
- TIMP2:
-
Tissue inhibitor of metalloproteinases 2
- Wnt:
-
Wingless-related integration site
- Wnt-5a:
-
Wingless-related integration site family member 5a
- Wnt-7a:
-
Wingless-related integration site family member 7a
References
Alarcón R, Ingaramo PI, Rivera OE, Dioguardi GH, Repetti MR, Demonte LD, Milesi MM, Varayoud J, Muñoz-de-Toro M, Luque EH (2019) Neonatal exposure to a glyphosate-based herbicide alters the histofunctional differentiation of the ovaries and uterus in lambs. Mol Cell Endocrinol 482:45–56. https://doi.org/10.1016/j.mce.2018.12.007
Altamirano GA, Delconte MB, Gomez AL, Alarcón R, Bosquiazzo VL, Luque EH, Muñoz-de-Toro M, Kass L (2017) Early postnatal exposure to endosulfan interferes with the normal development of the male rat mammary gland. Toxicol Lett 281:102–109. https://doi.org/10.1016/j.toxlet.2017.09.012
American Society of Ichthyologists and Herpetologists (ASIH) (2004) Guidelines for use of live amphibians and reptiles in field and laboratory research. In: Beaupre SJ, Jacobson ER, Lillywhite HL, Zamudio K (eds) Herpetological animal care and use commitee, 2nd edn. https://static1.squarespace.com/static/618bf11a71fcdf5398996eda/t/62506ab46b41e01343215dd7/1649437365012/IACUC_GuidelinesforUseofAmphibsReptiles2004.pdf. Accessed June 2023
Brew K, Nagase H (2010) The tissue inhibitors of metalloproteinases (TIMPs): an ancient family with structural and functional diversity. Biochem Biophys Acta 1803:55–71. https://doi.org/10.1016/j.bbamcr.2010.01.003
Brew K, Dinakarpandian D, Nagase H (2000) Tissue inhibitors of metalloproteinases: evolution, structure and function. Biochem Biophys Acta 1477:267–283. https://doi.org/10.1016/S0167-4838(99)00279-4
Canesini G, Galoppo GH, Tavalieri YE, Lazzarino GP, Stoker C, Luque EH, Ramos JG, Muñoz-de-Toro M (2023) Disruption of the developmental programming of the gonad of the broad snouted caiman (Caiman latirostris) after in ovo exposure to atrazine. Environ Sci Pollut 30:40132–40146. https://doi.org/10.1007/s11356-022-25104-z
Chang J, Fang W, Chen L, Zhang P, Zhang G, Zhang H, Liang J, Wang Q, Ma F (2022) Toxicological effects, environmental behaviors and remediation technologies of herbicide atrazine in soil and sediment: a comprehensive review. Chemosphere 307:136006. https://doi.org/10.1016/j.chemosphere.2022.136006
Cikos S, Bukovska A, Koppel J (2007) Relative quantification of mRNA: comparison of methods currently used for real-time PCR data analysis. BMC Mol Biol 8:113. https://doi.org/10.1186/1471-2199-8-113
Cooke PS, Spencer TE, Bartol FF, Hayashi K (2013) Uterine glands: development, function and experimental model systems. Mol Hum Reprod 19:547–558. https://doi.org/10.1093/molehr/gat031
Galoppo GH, Stoker C, Canesini G, Schierano-Marotti G, Durando M, Luque EH, Muñoz-de-Toro M (2016) Postnatal development and histofunctional differentiation of the oviduct in the broad-snouted caiman (Caiman latirostris). Gen Comp Endocrinol 236:42–53. https://doi.org/10.1016/j.ygcen.2016.07.001
Galoppo GH, Canesini G, Tavalieri YE, Stoker C, Kass L, Luque EH, Muñoz-de-Toro M (2017) Bisphenol A disrupts the temporal pattern of histofunctional changes in the female reproductive tract of Caiman latirostris. Gen Comp Endocrinol 254:75–85. https://doi.org/10.1016/j.ygcen.2017.09.021
Galoppo GH, Tavalieri YE, Schierano-Marotti G, Osti MR, Luque EH, Muñoz-de-Toro MM (2020) Long-term effects of in ovo exposure to an environmentally relevant dose of atrazine on the thyroid gland of Caiman latirostris. Environ Res 186:109410. https://doi.org/10.1016/j.envres.2020.109410
Gill SE, Kassim SY, Birkland TP, Parks WC (2010) Mouse models of MMP and TIMP function. Methods Mol Biol 622:31–52. https://doi.org/10.1007/978-1-60327-299-5_2
Hayes TB, Anderson LL, Beasley VR, de Solla SR, Iguchi T, Ingraham H, Kestemont P, Kniewald J, Kniewald Z, Langlois VS, Luque EH, McCoy KA, Muñoz-de-Toro M, Oka T, Oliveira CA, Orton F, Ruby S, Suzawa M, Tavera-Mendoza LE, Trudeau VL, Victor-Costa AB, Willingham E (2011) Demasculinization and feminization of male gonads by atrazine: consistent effects across vertebrate classes. J Steroid Biochem Mol Biol 127:64–73. https://doi.org/10.1016/j.jsbmb.2011.03.015
Ingaramo PI, Milesi MM, Schimpf MG, Ramos JG, Vigezzi L, Muñoz-de-Toro M, Luque EH, Varayoud J (2016) Endosulfan affects uterine development and functional differentiation by disrupting Wnt7a and β-catenin expression in rats. Mol Cell Endocrinol 425:37–47. https://doi.org/10.1016/j.mce.2016.02.011
Jamora C, DasGupta R, Kocieniewski P, Fuchs E (2003) Links between signal transduction, transcription and adhesion in epithelial bud development. Nature 422:317–322. https://doi.org/10.1038/nature01458
Lee KH, Choi EY, Hyun MS, Jang BI, Kim TN, Kim SW, Song SK, Kim JH, Kim JR (2007) Association of extracellular cleavage of E-cadherin mediated by MMP-7 with HGF-induced in vitro invasion in human stomach cancer cells. Eur Surg Res 39:208–215. https://doi.org/10.1159/000101452
Lesniak-Walentyn A, Hrabia A (2016) Expression and localization of matrix metalloproteinases (MMP-2, -7, -9) and their tissue inhibitors (TIMP-2, -3) in the chicken oviduct during maturation. Cell Tissue Res 364:185–197. https://doi.org/10.1007/s00441-015-2290-9
Luque EH, Muñoz-de-Toro M, Ramos JG (2018) Estrogenic agonist, In: Skinner MK, (Ed.) Encyclopedia of reproduction. Academic Press, Cambridge MA, US, Elsevier, pp 753–759
Milesi MM, Durando M, Lorenz V, Gastiazoro MP, Varayoud J (2020) Postnatal exposure to endosulfan affects uterine development and fertility. Mol Cell Endocrinol 511:110855. https://doi.org/10.1016/j.mce.2020.110855
Morris JA, Jordan CL, Breedlove SM (2004) Sexual differentiation of the vertebrate nervous system. Nat Neurosci 7:1034–1039. https://doi.org/10.1038/nn1325
National research council (2011) Guide for the care and use of laboratory animals, 8th ed. The National Academies Press, Washington DC, USA. https://doi.org/10.17226/12910
Portelinha TCG, Jahn GA, Hapon MB, Verdade LM, Piña CI (2015) Hormone levels and ultrasound evaluation of Caiman latirostris (Crocodylia, Alligatoridae) ovulation. S Am J Herpetol 10:23–31. https://doi.org/10.2994/SAJH-D-14-00030.1
Radcliffe JC (2002) Pesticide use in Australia. https://www.applied.org.au/wp-content/uploads/2019/01/pesticide-use-australia.pdf. Accessed, January 2023
Ramos-Fernández E, Tapia-Rojas C, Ramírez VT, Inestrosa NC (2019) Wnt-7a stimulates dendritic spine morphogenesis and PSD-95 expression through canonical signaling. Mol Neurobiol 56:1870–1882. https://doi.org/10.1007/s12035-018-1162-1
Rodríguez HA, Kass L, Varayoud J, Ramos JG, Ortega HH, Durando M, Muñoz-de-Toro M, Luque EH (2003) Collagen remodeling in the guinea-pig uterine cervix at term is associated with a decrease in progesterone receptor expression. Mol Hum Reprod 9:807–813. https://doi.org/10.1093/molehr/gag099
Rodríguez PM, Ondarza PM, Miglioranza KSB, Ramírez CL, Vera B, Muntaner C, Guiñazú NL (2023) Pesticides exposure in pregnant Argentinian women: potential relations with the residence areas and the anthropometric neonate parameters. Chemosphere 138790. https://doi.org/10.1016/j.chemosphere.2023.138790
Sathishkumar P, Mohan K, Ganesan AR, Govarthanan M, Yusoff ARM, Gu FL (2022) Persistence, toxicological effect and ecological issues of endosulfan – a review. J Hazard Mater 416:125779. https://doi.org/10.1016/j.jhazmat.2021.125779
Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682. https://doi.org/10.1038/nmeth.2019
Sebastian R, Raghavan SC (2015) Exposure to endosulfan can result in male infertility due to testicular atrophy and reduced sperm count. Cell Death Discov 1:15020. https://doi.org/10.1038/cddiscovery.2015.20
Shan L, Wang F, Zhai D, Meng X, Liu J, Lv X (2023) Matrix metalloproteinases induce extracellular matrix degradation through various pathways to alleviate hepatic fibrosis. Biomed Pharmacother 161:114472. https://doi.org/10.1016/j.biopha.2023.114472
Stoker C, Rey F, Rodríguez H, Ramos JG, Sirosky P, Larriera A, Luque EH, Muñoz-de-Toro M (2003) Sex reversal effects on Caiman latirostris exposed to environmentally relevant doses of the xenoestrogen bisphenol A. Gen Comp Endocrinol 133:287–296. https://doi.org/10.1016/S0016-6480(03)00199-0
Stoker C, Beldoménico PM, Bosquiazzo VL, Zayas MA, Rey F, Rodríguez H, Muñoz-de-Toro M, Luque EH (2008) Developmental exposure to endocrine disruptor chemicals alters follicular dynamics and steroid levels in Caiman latirostris. Gen Comp Endocrinol 156:603–612. https://doi.org/10.1016/j.ygcen.2008.02.011
Tavalieri YE, Galoppo GH, Canesini G, Truter JG, Ramos JG, Luque EH, Muñoz-de-Toro M (2019) The external genitalia in juvenile Caiman latirostris differ in hormone sex determinate-female from temperature sex determinate-female. General and Comparative Endocrinology 273:236–248. https://doi.org/10.1016/j.ygcen.2018.10.003
Tavalieri YE, Galoppo GH, Canesini G, Luque EH, Muñoz-de-Toro M (2020) Effects of agricultural pesticides on the reproductive system of aquatic wildlife species, with crocodilians as sentinel species. Mol Cell Endocrinol 518:110918. https://doi.org/10.1016/j.mce.2020.110918
Tavalieri YE, Alarcón R, Tschopp MV, Canesini G, Luque EH, Muñoz-de-Toro M, Galoppo GH (2021) Exposure to xenoestrogens alters the expression of key morphoregulatory proteins of oviduct adenogenesis in the broad-snouted caiman (Caiman latirostris). Aquat Toxicol 235:105817. https://doi.org/10.1016/j.aquatox.2021.105817
Tepekoy F, Akkoyunlu G, Demir R (2015) The role of Wnt signaling members in the uterus and embryo during pre-implantation and implantation. J Assist Reprod Genet 32:337–346. https://doi.org/10.1007/s10815-014-0409-7
Tian X, Liu Z, Niu B, Zhang J, Tan TK, Lee SR, Zhao Y, Harris DC, Zheng G (2011) E-cadherin/beta-catenin complex and the epithelial barrier. J Biomed Biotechnol 2011:567305
Van Doren SR (2015) Matrix metalloproteinase interactions with collagen and elastin. Matrix Biol 44–46:224–231. https://doi.org/10.1155/2011/567305
Van Opstal NV, Gabioud EA, Seehaus M, Pighini RJ, Repetti MR, Wilson MG, Wingeyer AB, Cuatrin A, Regaldo LM, Gagneten AM, Sasal MC (2023) Spatial distribution of pesticides in surface water of the Estacas stream (Argentine Espinal region) associated with crop production. Environ Sci Pollut Res. https://doi.org/10.1007/s11356-023-25373-2
Zampini R, Sari LM, Arganaraz ME, Fumuso FG, Barraza DE, Carretero MI, Apichela SA (2018) Mating induces production of MMP2 in the llama oviduct: analysis of MMP2 effect on SDen. Anim Reprod Sci 192:223–232. https://doi.org/10.1016/j.anireprosci.2018.03.016
Zayas MA, Rodríguez H, Galoppo GH, Stoker C, Durando M, Luque EH, Muñoz-de-Toro M (2011) Hematology and blood biochemistry of young healthy broad-snouted caimans (Caiman latirostris). J Herpetol 45:516–524. https://doi.org/10.1670/10-158.1
Acknowledgements
We thank Juan Grant and Walter Nykolajczuk for technical assistance and animal care. Field work was done in collaboration with ‘‘Reserva Natural El Cachapé,” Chaco, Argentina. YT is a doctoral fellow and RA is a post-doctoral fellow of CONICET, MVT is an assistant professional of the supporting research career and EHL is a career investigator of CONICET.
Funding
This study was supported by grants from the Argentine National Agency for the Promotion of Science and Technology (ANPCyT; PICT2016-0656), the Universidad Nacional del Litoral (CAI + D Program, 1–15, 2016), and the National Scientific and Technical Research Council of Argentina (CONICET; PIP2021-2023 Cod. 1220200101387CO).
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All authors contributed to the study conception and design. Material preparation and data collection and analysis were performed by Yamil Ezequiel Tavalieri. The first draft of the manuscript was written by Yamil Ezequiel Tavalieri and Ramiro Alarcón, and all the authors commented on previous versions of the manuscript. Yamil Ezequiel Tavalieri and Ramiro Alarcón contributed with visualization as well. María Virginia Tschopp and Germán Hugo Galoppo supplied methodological support. The review and edition of the final manuscript were performed by Enrique Hugo Luque, Mónica Muñoz-de-Toro, and Germán Hugo Galoppo. Enrique Hugo Luque and Mónica Muñoz-de-Toro contributed with funding acquisition. All authors read and approved the final manuscript.
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The protocols used in this work were previously authorized by the Institutional Committee of Bioethics in Animal Care and Use of the Universidad Nacional del Litoral, Santa Fe, Argentina. All procedures and techniques used for egg collection, embryo handling, and sampling were performed according to the guidelines of the American Society of Ichthyologists and Herpetologists (ASIH, 2004). All animal experiments were carried out in accordance with the Guide for the Care and Use of Laboratory Animals of the National Research Council (USA) (IPCS, 2002).
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Tavalieri, Y., Alarcón, R., Tschopp, M. et al. Exposure to atrazine and endosulfan alters oviductal adenogenesis in the broad-snouted caiman (Caiman latirostris). Environ Sci Pollut Res 31, 35927–35937 (2024). https://doi.org/10.1007/s11356-024-33662-7
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DOI: https://doi.org/10.1007/s11356-024-33662-7