Hostname: page-component-78c5997874-mlc7c Total loading time: 0 Render date: 2024-11-16T13:24:21.809Z Has data issue: false hasContentIssue false
  • English
  • Français

Cellular Evidence of CD63-Enriched Exosomes and Multivesicular Bodies within the Seminiferous Tubule during the Spermatogenesis of Turtles

Published online by Cambridge University Press:  22 November 2019

Imran Tarique ,
Abdul Haseeb ,
Xuebing Bai ,
Wenqian Li ,
Ping Yang ,
Yufei Huang ,
Sheng Yang ,
Mengdi Xu ,
Yue Zhang  and
Waseem Ali Vistro
...Show all authors
Imran Tarique
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Abdul Haseeb
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China Faculty of Veterinary and Animal Sciences, University of Poonch Rawalakot, Azad Kashmir, Pakistan
Xuebing Bai
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Wenqian Li
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Ping Yang
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Yufei Huang
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Sheng Yang
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Mengdi Xu
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Yue Zhang
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Waseem Ali Vistro
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Surfaraz Ali Fazlani
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
Qiusheng Chen*
Affiliation:
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province210095, China
*
*Author for correspondence: Qiusheng Chen, E-mail: [email protected]
Get access

Abstract

The seminiferous tubule (ST) is the location of spermatogenesis, where mature spermatozoa are produced with the assistance of Sertoli cells. The role of extracellular vesicles in the direct communication between Sertoli-germ cells in the ST is still not fully understood. In this study, we reported multivesicular bodies (MVBs) and their source of CD63-enriched exosomes by light and ultrastructure microscopy during the reproductive phases of turtles. Strong CD63 immunopositivity was detected at the basal region in the early and luminal regions of the ST during late spermatogenesis by immunohistochemistry (IHC), immunofluorescence (IF), and western blot (WB) analysis. Labeling of CD63 was detected in the Sertoli cell cytoplasmic processes that surround the developing germ cells during early spermatogenesis and in the lumen of the ST with elongated spermatids during late spermatogenesis. Furthermore, ultrastructure analysis confirmed the existence of numerous MVBs in the Sertoli cell prolongations that surround the round and primary spermatogonia during acrosome biogenesis and with the embedded heads of spermatids in the cytoplasm of Sertoli cells. Additionally, in spermatids, Chrysanthemum flower centers (CFCs) generated isolated membranes involved in MVBs and autophagosome formation, and their fusion to form amphiosomes was also observed. Additionally, autophagy inhibition by 3-methyladenine (after 24 h) increased CD63 protein signals during late spermatogenesis, as detected by IF and WB. Collectively, our study found MVBs and CD63 rich exosomes within the Sertoli cells and their response to autophagy inhibition in the ST during the spermatogenesis in the turtle.

Keywords

autophagyCD63 exosomemultivesicular bodiesSertoli cellspermatogenesisturtle
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 26 , Issue 1 , February 2020 , pp. 148 - 156
Copyright
Copyright © Microscopy Society of America 2019

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

a

The authors contributed equally to this work.

References

Abels, ER & Breakefield, XO (2016). Introduction to extracellular vesicles: Biogenesis, RNA cargo selection, content, release, and uptake. Cell Mol Neurobiol 36(3), 301–312.CrossRefGoogle ScholarPubMed
Abid Hussein, MN, Nieuwland, R, Hau, CM, Evers, LM, Meesters, EW & Sturk, A (2005). Cell-derived microparticles contain caspase 3 in vitro and in vivo. J Thromb Haemost 3(5), 888–896.CrossRefGoogle ScholarPubMed
Abu Elhija, M, Lunenfeld, E, Schlatt, S & Huleihel, M (2012). Differentiation of murine male germ cells to spermatozoa in a soft agar culture system. Asian J Androl 14(2), 285–293.CrossRefGoogle Scholar
Bevers, EM, Comfurius, P, Van Rijn, JLML & Hemker, HC (1982). Generation of prothrombin-converting activity and the exposure of phosphatidylserine at the outer surface of platelets. Eur J Biochem 122(2), 429–436.CrossRefGoogle ScholarPubMed
Bian, X, Gandahi, JA, Liu, Y, Yang, P, Liu, Y, Zhang, L, Zhang, Q & Chen, Q (2013). The ultrastructural characteristics of the spermatozoa stored in the cauda epididymidis in Chinese soft-shelled turtle Pelodiscus sinensis during the breeding season. Micron 44, 202–209.CrossRefGoogle ScholarPubMed
Boyiadzis, M & Whiteside, TL (2015). Information transfer by exosomes: A new frontier in hematologic malignancies. Blood Rev 29(5), 281–290.CrossRefGoogle ScholarPubMed
Chen, H, Huang, Y, Yang, P, Liu, T, Ahmed, N, Wang, L, Wang, T, Bai, X, Haseeb, A & Chen, Q (2019). Lipophagy contributes to long-term storage of spermatozoa in the epididymis of the Chinese soft-shelled turtle Pelodiscus sinensis. Reprod Fert Develop 31(4), 774–786.CrossRefGoogle ScholarPubMed
Chen, H, Yang, P, Chu, X, Huang, Y, Liu, T, Zhang, Q, Li, Q, Hu, L, Waqas, Y, Ahmed, N & Chen, Q (2016). Cellular evidence for nano-scale exosome secretion and interactions with spermatozoa in the epididymis of the Chinese soft-shelled turtle, Pelodiscus sinensis. Oncotarget 7(15), 19242–19250.Google ScholarPubMed
Chen, Y, Zhou, Y, Wang, XT, Qian, WP & Han, XD (2013). Microcystin-LR induces autophagy and apoptosis in rat Sertoli cells in vitro. Toxicon 76, 84–93.CrossRefGoogle ScholarPubMed
Cheng, CY & Mruk, DD (2002). Cell junction dynamics in the testis: Sertoli-germ cell interactions and male contraceptive development. Physiol Rev 82(4), 825–874.CrossRefGoogle ScholarPubMed
Cheng, CY & Mruk, DD (2010). The biology of spermatogenesis: The past, present and future. Philos Trans R Soc Lond B Biol Sci 365(1546), 1459–1463.CrossRefGoogle ScholarPubMed
Da Ros, M, Hirvonen, N, Olotu, O, Toppari, J & Kotaja, N (2015). Retromer vesicles interact with RNA granules in haploid male germ cells. Mol Cell Endocrinol 401, 73–83.CrossRefGoogle ScholarPubMed
Fader, CM & Colombo, MI (2009). Autophagy and multivesicular bodies: Two closely related partners. Cell Death Differ 16(1), 70–78.CrossRefGoogle ScholarPubMed
Hanson, PI & Cashikar, A (2012). Multivesicular body morphogenesis. Ann Rev Cell Dev Biol 28(1), 337–362.CrossRefGoogle ScholarPubMed
Haseeb, A, Tarique, I, Bai, X, Yang, P, Ali Vistro, W, Huang, Y, Ali Fazllani, S, Ahmed, Z & Chen, Q (2019). Inhibition of autophagy impairs acrosome and mitochondrial crista formation during spermiogenesis in turtle: Ultrastructural evidence. Micron 121, 84–89.CrossRefGoogle ScholarPubMed
Hess, RA & França, LR (2005). Chapter 3 - Structure of the Sertoli cell. In Sertoli Cell Biology, Skinner, MK & Griswold, MD (Eds.), pp. 19–40. San Diego: Academic Press.CrossRefGoogle Scholar
Huang, Y, Yang, P, Liu, T, Chen, H, Chu, X, Ahmad, N, Zhang, Q, Li, Q, Hu, L, Liu, Y & Chen, Q (2016). Subcellular evidence for biogenesis of autophagosomal membrane during spermiogenesis in vivo. Front Physiol 7, 470.CrossRefGoogle ScholarPubMed
Kapsogeorgou, EK, Abu-Helu, RF, Moutsopoulos, HM & Manoussakis, MN (2005). Salivary gland epithelial cell exosomes: A source of autoantigenic ribonucleoproteins. Arthritis Rheum 52(5), 1517–1521.CrossRefGoogle ScholarPubMed
Kosaka, N, Iguchi, H & Ochiya, T (2010). Circulating microRNA in body fluid: A new potential biomarker for cancer diagnosis and prognosis. Cancer Sci 101(10), 2087–2092.CrossRefGoogle ScholarPubMed
Lee, NPY, Mruk, DD, Wong, CH & Cheng, CY (2005). Regulation of Sertoli-germ cell adherens junction dynamics in the testis via the nitric oxide synthase (NOS)/cGMP/protein kinase G (PRKG)/β-catenin (CATNB) signaling pathway: An in vitro and in vivo study. Biol Reprod 73(3), 458–471.CrossRefGoogle Scholar
Ma, Y, Yang, HZ, Xu, LM, Huang, YR, Dai, HL & Kang, XN (2015). Testosterone regulates the autophagic clearance of androgen binding protein in rat Sertoli cells. Scientific Reports 5.CrossRefGoogle Scholar
Metzelaar, MJ, Wijngaard, PL, Peters, PJ, Sixma, JJ, Nieuwenhuis, HK & Clevers, HC (1991). CD63 antigen. A novel lysosomal membrane glycoprotein, cloned by a screening procedure for intracellular antigens in eukaryotic cells. J Biol Chem 266(5), 3239–3245.Google ScholarPubMed
Mittelbrunn, M, Gutierrez-Vazquez, C, Villarroya-Beltri, C, Gonzalez, S, Sanchez-Cabo, F, Gonzalez, MA, Bernad, A & Sanchez-Madrid, F (2011). Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells. Nat Commun 2, 282.CrossRefGoogle Scholar
Mizushima, N, Levine, B, Cuervo, AM & Klionsky, DJ (2008). Autophagy fights disease through cellular self-digestion. Nature 451(7182), 1069–1075.CrossRefGoogle ScholarPubMed
Ozturk, N, Steger, K & Schagdarsurengin, U (2017). The impact of autophagy in spermiogenesis. Asian J Androl 19(6), 617–618.Google ScholarPubMed
Poliakov, A, Spilman, M, Dokland, T, Amling, CL & Mobley, JA (2009). Structural heterogeneity and protein composition of exosome-like vesicles (prostasomes) in human semen. Prostate 69(2), 159–167.CrossRefGoogle Scholar
Pols, MS & Klumperman, J (2009). Trafficking and function of the tetraspanin CD63. Exp Cell Res 315(9), 1584–1592.CrossRefGoogle ScholarPubMed
Raposo, G & Stoorvogel, W (2013). Extracellular vesicles: Exosomes, microvesicles, and friends. J Cell Biol 200(4), 373–383.CrossRefGoogle ScholarPubMed
Rieu, S, Geminard, C, Rabesandratana, H, Sainte-Marie, J & Vidal, M (2000). Exosomes released during reticulocyte maturation bind to fibronectin via integrin alpha4beta1. Eur J Biochem 267(2), 583–590.CrossRefGoogle ScholarPubMed
Sharma, R & Agarwal, A (2011). Spermatogenesis: An overview. In Sperm Chromatin: Biological and Clinical Applications in Male Infertility and Assisted Reproduction, Zini, A & Agarwal, A (Eds.), pp. 19–44. New York, NY: Springer New York.CrossRefGoogle Scholar
Simons, M & Raposo, G (2009). Exosomes–vesicular carriers for intercellular communication. Curr Opin Cell Biol 21(4), 575–581.CrossRefGoogle ScholarPubMed
Ujjan, N, Liu, Y, Chen, H, Yang, P, Waqas, Y, Liu, T, Gandahi, DJ, Huang, Y, Wang, L, Song, X, Rajput, I, Wang, T & Chen, Q (2016). Novel cellular evidence of lipophagy within the Sertoli cells during spermatogenesis in the turtle. Aging 9, 41–51.Google Scholar
Valadi, H, Ekstrom, K, Bossios, A, Sjostrand, M, Lee, JJ & Lotvall, JO (2007). Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 9(6), 654–U672.CrossRefGoogle ScholarPubMed
van der Pol, E, Boing, AN, Harrison, P, Sturk, A & Nieuwland, R (2012). Classification, functions, and clinical relevance of extracellular vesicles. Pharmacol Rev 64(3), 676–705.CrossRefGoogle ScholarPubMed
van Niel, G, Raposo, G, Candalh, C, Boussac, M, Hershberg, R, Cerf-Bensussan, N & Heyman, M (2001). Intestinal epithelial cells secrete exosome-like vesicles. Gastroenterology 121(2), 337–349.CrossRefGoogle ScholarPubMed
Voorhout, WF, Veenendaal, T, Haagsman, HP, Weaver, TE, Whitsett, JA, van Golde, LM & Geuze, HJ (1992). Intracellular processing of pulmonary surfactant protein B in an endosomal/lysosomal compartment. Am J Physiol 263(4 Pt 1), L479–L486.Google Scholar
Voorhout, WF, Weaver, TE, Haagsman, HP, Geuze, HJ & Van Golde, LM (1993). Biosynthetic routing of pulmonary surfactant proteins in alveolar type II cells. Microsc Res Tech 26(5), 366–373.CrossRefGoogle ScholarPubMed
Whiteside, TL (2005). Tumour-derived exosomes or microvesicles: Another mechanism of tumour escape from the host immune system? Br J Cancer 92(2), 209–211.CrossRefGoogle ScholarPubMed
Xu, D & Tahara, H (2013). The role of exosomes and microRNAs in senescence and aging. Adv Drug Deliver Rev 65(3), 368–375.CrossRefGoogle ScholarPubMed
Xu, J, Camfield, R & Gorski, SM (2018). The interplay between exosomes and autophagy – Partners in crime. J Cell Sci 131(15). doi:10.1242/jcs.215210.CrossRefGoogle ScholarPubMed
Yang, P, Ahmed, N, Wang, LL, Chen, H, Waqas, Y, Liu, TF, Haseeb, A, Bangulzai, N, Huang, YF & Chen, QS (2017). In vivo autophagy and biogenesis of autophagosomes within male haploid cells during spermiogenesis. Oncotarget 8(34), 56791–56801.Google ScholarPubMed