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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Histology, ultrastructure, and differential gene expression in relation to seasonal sperm storage in the oviduct of the Chinese alligator, Alligator sinensis

Haitao Nie https://orcid.org/0000-0002-2332-6391 A # , Yue Wen A , Ruidong Zhang B # , Mengjuan Wu A , Yuqian Zhang A , Yunlu Xu A , Jixiang Zhan A and Xiaobing Wu A *
+ Author Affiliations
- Author Affiliations

A Key Laboratory for Conservation and Use of Important Biological Resources of Anhui Province, College of Life Sciences, Anhui Normal University, Wuhu, Anhui 241000, China.

B College of Life Sciences, Inner Mongolia Normal University, Hohhot, Inner Mongolia 010022, China.

* Correspondence to: wuxb@mail.ahnu.edu
# These authors contributed equally to this paper

Handling Editor: Tod Fullston

Reproduction, Fertility and Development 34(12) 819-832 https://doi.org/10.1071/RD21191
Published online: 17 May 2022

© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND)

Abstract

Although oviductal sperm storage are essential steps in reproduction for female animals with internal fertilisation, no systematic study on the identification of genes involving sperm storage has been performed in crocodilian species. In the present research, the relationship between morphological variation related to sperm storage in the oviduct and gene expression patterns derived from RNA sequencing analyses between active period (AP), breeding period (BP), and hibernation period (HP) were investigated. The corresponding results indicated that sperm were observed not only in the ciliated cells within infundibulum and mucosal layer of uterus during BP, but also been detected in the spermatosperm storage tube (SST) in the anterior uterus at HP stage. The further transmission electron microscopy analysis indicated that the differences in the number and activity of the secretory cells likely to attributed to the seasonal variation of microenvironment related to the sperm storage. Based on the RNA-sequecing, 13 147 DEGs related to the Peroxisome proliferator-activated receptors (PPARs) and FOXO signalling were identified, including these, the down-regulated ATG12 and BCL2L11 in the HP group may thus constitute an important point of convergence between autophagy and apoptosis involving the FOXO1 pathway. The genes involved in the PPARs pathway might modulate the immune response and thereby contribute to prolong the life span of stored spermatozoa in Alligator sinensis. The outcomes of this study provide fundamental insights into the mechanism of sperm storage in A. sinensis.

Keywords: Chinese alligator, fertility, microscopy, oviduct, reproductive cycles, RNA sequencing, sperm storage, ultrastructure.


References

Abe, H (1996). The mammalian oviductal epithelium: regional variations in cytological and functional aspects of the oviductal secretory cells. Histology and Histopathology 11, 743–768.
| 8839764PubMed |

Abe, H, Ookata, K, Abe, M, and Oikawa, T (1992). Immunological characterization of oviductal glycoproteins associated with the zona pellucida of the golden hamster egg. Journal of Experimental Zoology 262, 209–218.
Immunological characterization of oviductal glycoproteins associated with the zona pellucida of the golden hamster egg.Crossref | GoogleScholarGoogle Scholar | 1583463PubMed |

Abughrien, BM, and Dore, MAP (2000). Ciliogenesis in the uterine tube of control and superovulated heifers. Cells, Tissues, Organs 166, 338–348.
Ciliogenesis in the uterine tube of control and superovulated heifers.Crossref | GoogleScholarGoogle Scholar | 10867435PubMed |

Aitken, RNC, and Solomon, SE (1976). Observations on the ultrastructure of the oviduct of the Costa Rican green turtle (Chelonia mydas L.). Journal of Experimental Marine Biology and Ecology 21, 75–90.
Observations on the ultrastructure of the oviduct of the Costa Rican green turtle (Chelonia mydas L.).Crossref | GoogleScholarGoogle Scholar |

Al Rawi, S, Louvet-Vallée, S, Djeddi, A, Sachse, M, Culetto, E, Hajjar, C, Boyd, L, Legouis, R, and Galy, V (2011). Postfertilization autophagy of sperm organelles prevents paternal mitochondrial DNA transmission. Science 334, 1144–1147.
Postfertilization autophagy of sperm organelles prevents paternal mitochondrial DNA transmission.Crossref | GoogleScholarGoogle Scholar | 22033522PubMed |

Audo, R, Deckert, V, Daien, C, Che, H, Elhmioui, J, Pais de Barros, J-P, Lemaire, S, Desrumaux, C, Hahne, M, Combe, B, Lagrost, L, and Morel, J (2016). OP0212 phospholipid transfer protein (PLTP): a link between inflammation and lipids in rheumatoid arthritis? Annals of the Rheumatic Diseases 75, 137.1–137.
OP0212 phospholipid transfer protein (PLTP): a link between inflammation and lipids in rheumatoid arthritis?Crossref | GoogleScholarGoogle Scholar |

Bagwill, A, Sever, DM, and Elsey, RM (2009). Seasonal variation of the oviduct of the american alligator, Alligator mississippiensis (Reptilia: Crocodylia). Journal of Morphology 270, 702–713.
Seasonal variation of the oviduct of the american alligator, Alligator mississippiensis (Reptilia: Crocodylia).Crossref | GoogleScholarGoogle Scholar | 19206152PubMed |

Bakst, MR (1998). Structure of the avian oviduct with emphasis on sperm storage in poultry. Journal of Experimental Zoology Part A: Ecological Genetics and Physiology 282, 618–626.
Structure of the avian oviduct with emphasis on sperm storage in poultry.Crossref | GoogleScholarGoogle Scholar |

Bareither, ML, and Verhage, HG (1981). Control of the secretory cell cycle in cat oviduct by estradiol and progesterone. American Journal of Anatomy 162, 107–118.
Control of the secretory cell cycle in cat oviduct by estradiol and progesterone.Crossref | GoogleScholarGoogle Scholar | 7304467PubMed |

Boisvieux-Ulrich, E, and Sandoz, D (1991). Determination of ciliary polarity precedes differentiation in the epithelial cells of quail oviduct. Biology of the Cell 72, 3–14.
Determination of ciliary polarity precedes differentiation in the epithelial cells of quail oviduct.Crossref | GoogleScholarGoogle Scholar | 1756309PubMed |

Chen, B, and Wang, C (1984). Artificial reproduction of Alligator sinensis. (In Chinese with English abstract.). Acta Pharmacologica Sinica 3, 49–54.

Das, SC, Isobe, N, and Yoshimura, Y (2008). Mechanism of prolonged sperm storage and sperm survivability in hen oviduct: a review. American Journal of Reproductive Immunology 60, 477–481.
Mechanism of prolonged sperm storage and sperm survivability in hen oviduct: a review.Crossref | GoogleScholarGoogle Scholar | 19032608PubMed |

Das, SC, Isobe, N, and Yoshimura, Y (2009). Changes in the expression of interleukin-1β and lipopolysaccharide-induced TNF factor in the oviduct of laying hens in response to artificial insemination. Reproduction 137, 527–536.
Changes in the expression of interleukin-1β and lipopolysaccharide-induced TNF factor in the oviduct of laying hens in response to artificial insemination.Crossref | GoogleScholarGoogle Scholar | 18539728PubMed |

Davenport, M (1995). Evidence of possible sperm storage in the caiman, Paleosuchus palpebrosus. Herpetological Review 26, 14–15.

Eun, CS, Han, DS, Lee, SH, Paik, CH, Chung, YW, Lee, J, and Hahm, JS (2006). Attenuation of colonic inflammation by PPARγ in intestinal epithelial cells: effect on toll-like receptor pathway. Digestive Diseases and Sciences 51, 693–697.
Attenuation of colonic inflammation by PPARγ in intestinal epithelial cells: effect on toll-like receptor pathway.Crossref | GoogleScholarGoogle Scholar | 16614990PubMed |

Gallardo Bolaños, JM, Miró Morán, Á, Balao da Silva, CM, Morillo Rodríguez, A, Plaza Dávila, M, Aparicio, IM, Tapia, JA, Ortega Ferrusola, C, and Peña, FJ (2012). Autophagy and apoptosis have a role in the survival or death of stallion spermatozoa during conservation in refrigeration. PLoS ONE 7, e30688.
Autophagy and apoptosis have a role in the survival or death of stallion spermatozoa during conservation in refrigeration.Crossref | GoogleScholarGoogle Scholar | 22292020PubMed |

Gist, DH, Bagwill, A, Lance, V, Sever, DM, and Elsey, RM (2008). Sperm storage in the oviduct of the American alligator. Journal of Experimental Zoology Part A: Ecological Genetics and Physiology 309, 581–587.
Sperm storage in the oviduct of the American alligator.Crossref | GoogleScholarGoogle Scholar |

Guillette, LJ, Fox, SL, and Palmer, BD (1989). Oviductal morphology and egg shelling in the oviparous lizards Crotaphytus collaris and Eumeces obsoletus. Journal of Morphology 201, 145–159.
Oviductal morphology and egg shelling in the oviparous lizards Crotaphytus collaris and Eumeces obsoletus.Crossref | GoogleScholarGoogle Scholar | 29865655PubMed |

Han, X, Li, Z, Li, M, Bao, H, and Chen, Q (2008). Ultrastructure of anterior uterus of the oviduct and the stored sperm in female soft-shelled turtle, Trionyx sinensis. Anatomical Record 291, 335–351.
Ultrastructure of anterior uterus of the oviduct and the stored sperm in female soft-shelled turtle, Trionyx sinensis.Crossref | GoogleScholarGoogle Scholar |

Hawk, HW, Conley, HH, Wall, RJ, and Whitaker, RO (1988). Fertilization rates in superovulating cows after deposition of semen on the infundibulum, near the uterotubal junction or after insemination with high numbers of sperm. Theriogenology 29, 1131–1142.
Fertilization rates in superovulating cows after deposition of semen on the infundibulum, near the uterotubal junction or after insemination with high numbers of sperm.Crossref | GoogleScholarGoogle Scholar | 16726436PubMed |

Hunter, RHF, Fléchon, B, and Fléchon, JE (1991). Distribution, morphology and epithelial interactions of bovine spermatozoa in the oviduct before and after ovulation: a scanning electron microscope study. Tissue and Cell 23, 641–656.
Distribution, morphology and epithelial interactions of bovine spermatozoa in the oviduct before and after ovulation: a scanning electron microscope study.Crossref | GoogleScholarGoogle Scholar |

Kadivar, A, Khoei, HH, Hassanpour, H, Ghanaei, H, Golestanfar, A, Mehraban, H, Davoodian, N, and Tafti, RD (2016). Peroxisome proliferator-activated receptors (PPARα, PPARγ and PPARβ/δ) gene expression profile on ram spermatozoa and their relation to the sperm motility. Veterinary Research Forum 7, 27–34.
| 27226884PubMed |

Kawashima, M, Takahashi, T, Kamiyoshi, M, and Tanaka, K (1996). Effects of progesterone, estrogen, and androgen on progesterone receptor binding in hen oviduct uterus (shell gland). Poultry Science 75, 257–260.
Effects of progesterone, estrogen, and androgen on progesterone receptor binding in hen oviduct uterus (shell gland).Crossref | GoogleScholarGoogle Scholar | 8833380PubMed |

Kucera, AC, and Heidinger, BJ (2018). Avian semen collection by cloacal massage and isolation of DNA from sperm. Journal of Visualized Experiments , e55324.
Avian semen collection by cloacal massage and isolation of DNA from sperm.Crossref | GoogleScholarGoogle Scholar |

Larsen, RE, Cardeilhac, PT, and Lane, T (1984). Semen extenders for artificial insemination in the American alligator. Aquaculture 42, 141–149.
Semen extenders for artificial insemination in the American alligator.Crossref | GoogleScholarGoogle Scholar |

Li, Q, Hu, L, Yang, P, Zhang, Q, and Chen, Q (2015). Expression of TLR2/4 in the sperm-storing oviduct of the Chinese soft-shelled turtle Pelodiscus sinensis during hibernation season. Ecology and Evolution 5, 4466–4479.
Expression of TLR2/4 in the sperm-storing oviduct of the Chinese soft-shelled turtle Pelodiscus sinensis during hibernation season.Crossref | GoogleScholarGoogle Scholar | 26664692PubMed |

Liu, T, Yang, P, Chen, H, Huang, Y, Liu, Y, Waqas, Y, Ahmed, N, Chu, X, and Chen, Q (2016). Global analysis of differential gene expression related to long-term sperm storage in oviduct of Chinese Soft-Shelled Turtle Pelodiscus sinensis. Scientific Reports 6, 33296.
Global analysis of differential gene expression related to long-term sperm storage in oviduct of Chinese Soft-Shelled Turtle Pelodiscus sinensis.Crossref | GoogleScholarGoogle Scholar | 27628424PubMed |

Loos, B, Lochner, A, and Engelbrecht, A-M (2011). Autophagy in heart disease: a strong hypothesis for an untouched metabolic reserve. Medical Hypotheses 77, 52–57.
Autophagy in heart disease: a strong hypothesis for an untouched metabolic reserve.Crossref | GoogleScholarGoogle Scholar | 21482032PubMed |

Luo, S, and Rubinsztein, DC (2013). BCL2L11/BIM: a novel molecular link between autophagy and apoptosis. Autophagy 9, 104–105.
BCL2L11/BIM: a novel molecular link between autophagy and apoptosis.Crossref | GoogleScholarGoogle Scholar | 23064249PubMed |

Madekurozwa, M-C (2013). An immunohistochemical study of the oviduct in the domestic fowl (Gallus domesticus). Anatomia Histologia Embryologia 42, 48–56.
An immunohistochemical study of the oviduct in the domestic fowl (Gallus domesticus).Crossref | GoogleScholarGoogle Scholar | 22671283PubMed |

Mastroianni L, Stambaugh R (1974) Secretory function of the primate oviduct. In ‘Physiology and genetics of reproduction’. Basic life sciences. (Eds EM Coutinho, F Fuchs) vol. 4. pp. 25–34. (Springer: Boston, MA).
| Crossref |

Nie, H, Zhang, R, Yu, X, Zhang, Y, Yan, P, Li, E, Wang, R, and Wu, X (2021). Molecular cloning, immunological characterization, and expression analysis of gonadotropin-releasing hormone (GnRH) in the brain of the Chinese alligator during different stages of reproductive cycle. Gene 789, 145672.
Molecular cloning, immunological characterization, and expression analysis of gonadotropin-releasing hormone (GnRH) in the brain of the Chinese alligator during different stages of reproductive cycle.Crossref | GoogleScholarGoogle Scholar | 33882325PubMed |

Odor, DL, and Blandau, RJ (1973). EGG transport over the fimbrial surface of the rabbit oviduct under experimental conditions. Fertility and Sterility 24, 292–300.
EGG transport over the fimbrial surface of the rabbit oviduct under experimental conditions.Crossref | GoogleScholarGoogle Scholar | 4694504PubMed |

Palladino, MA, Johnson, TA, Gupta, R, Chapman, JL, and Ojha, P (2007). Members of the toll-like receptor family of innate immunity pattern-recognition receptors are abundant in the male rat reproductive tract. Biology of Reproduction 76, 958–964.
Members of the toll-like receptor family of innate immunity pattern-recognition receptors are abundant in the male rat reproductive tract.Crossref | GoogleScholarGoogle Scholar | 17314314PubMed |

Palmer, B, and Guillette, LJ (1988). Histology and functional morphology of the female reproductive tract of the tortoise Gopherus polyphemus. Developmental Dynamics 183, 200–211.
Histology and functional morphology of the female reproductive tract of the tortoise Gopherus polyphemus.Crossref | GoogleScholarGoogle Scholar |

Palmer, B, and Guillette, LJ (1992). Alligators provide evidence for the Archosaurian mode of oviparity. Biology of Reproduction 46, 39–47.
Alligators provide evidence for the Archosaurian mode of oviparity.Crossref | GoogleScholarGoogle Scholar | 1547315PubMed |

Rabbani, SI, Devi, K, and Khanam, S (2010). Pioglitazone, a PPAR-gamma ligand inhibited the nicotinamide-streptozotocin induced sperm abnormalities in type-2 diabetic Wistar rats. Pakistan Journal of Pharmaceutical Sciences 23, 326–331.
| 20566448PubMed |

Rubinstein, AD, Eisenstein, M, Ber, Y, Bialik, S, and Kimchi, A (2011). The autophagy protein Atg12 associates with antiapoptotic Bcl-2 family members to promote mitochondrial apoptosis. Molecular Cell 44, 698–709.
The autophagy protein Atg12 associates with antiapoptotic Bcl-2 family members to promote mitochondrial apoptosis.Crossref | GoogleScholarGoogle Scholar | 22152474PubMed |

Sasanami, T, Matsuzaki, M, Mizushima, S, and Hiyama, G (2013). Sperm storage in the female reproductive tract in birds. Journal of Reproduction and Development 59, 334–338.
Sperm storage in the female reproductive tract in birds.Crossref | GoogleScholarGoogle Scholar | 23965601PubMed |

Sever, DM, and Ryan, TJ (1999). Ultrastructure of the reproductive system of the black swamp snake (Seminatrix pygaea): Part I. Evidence for oviducal sperm storage. Journal of Morphology 241, 1–18.
Ultrastructure of the reproductive system of the black swamp snake (Seminatrix pygaea): Part I. Evidence for oviducal sperm storage.Crossref | GoogleScholarGoogle Scholar | 10398321PubMed |

Swampillai, AL, Salomoni, P, and Short, SC (2012). The role of autophagy in clinical practice. Clinical Oncology 24, 387–395.
The role of autophagy in clinical practice.Crossref | GoogleScholarGoogle Scholar | 22032864PubMed |

Uller, T, Schwartz, T, Koglin, T, and Olsson, M (2013). Sperm storage and sperm competition across ovarian cycles in the dragon lizard, Ctenophorus fordi. Journal of Experimental Zoology Part A Ecological Genetics and Physiology 319, 404–408.
Sperm storage and sperm competition across ovarian cycles in the dragon lizard, Ctenophorus fordi.Crossref | GoogleScholarGoogle Scholar | 23744523PubMed |

Wang, Z, and Zhang, SY (2005). Sperm storage in the bat, myotis ricketti. Progress in Natural Science 15, 1011–1014.

Wang, Z, Shi, QX, Wang, YN, Wang, YL, and Zhang, SY (2008). Epididymal sperm storage in Rickett’s big-footed bat (Myotis ricketti). Acta Chiropterologica 10, 161–167.
Epididymal sperm storage in Rickett’s big-footed bat (Myotis ricketti).Crossref | GoogleScholarGoogle Scholar |

Wun, IC, and Huang, JC (2008). Enhancement of sperm feritilization capability by peroxisome proliferation-activated receptor (PPAR) γ agonist. Fertility and Sterility 90, S318–S318.
Enhancement of sperm feritilization capability by peroxisome proliferation-activated receptor (PPAR) γ agonist.Crossref | GoogleScholarGoogle Scholar |

Yamanaka, M, Ishikawa, T, Griep, A, Axt, D, Kummer, MP, and Heneka, MT (2012). PPARγ/RXRα-induced and CD36-mediated microglial amyloid-β phagocytosis results in cognitive improvement in amyloid precursor protein/presenilin 1 mice. Journal of Neuroscience 32, 17321–17331.
PPARγ/RXRα-induced and CD36-mediated microglial amyloid-β phagocytosis results in cognitive improvement in amyloid precursor protein/presenilin 1 mice.Crossref | GoogleScholarGoogle Scholar | 23197723PubMed |