Three-dimensional analysis and in vivo imaging for sperm release and transport in the murine seminiferous tubule

in Reproduction
View More View Less
  • 1 Department of Anatomy, Tokyo Medical University, Tokyo, Japan
  • | 2 Department of Histology and Cell Biology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan
  • | 3 The Hakubi Center/Graduate School of Biostudies, Kyoto University, Kyoto, Japan
  • | 4 Japan Science and Technology Agency, PRESTO, Kawaguchi, Japan

Correspondence should be addressed to T Omotehara; Email: omote@tokyo-med.ac.jp
Restricted access

USD  $0.00
USD  $0.00

USD  $0.00
USD  $0.00

USD  $0.00
USD  $0.00

USD  $0.00
USD  $0.00

USD  $0.00
USD  $0.00

In Brief

Spermatozoa are released from Sertoli cells and flow in the seminiferous tubule to the rete testis. Our results suggest that the luminal flow in the tubules is repeatedly reversed and that this physical force helps spermatozoa release from the Sertoli cells.

Abstract

Spermatozoa released from Sertoli cells must be transported to the epididymis. However, the mechanism of the luminal flow in seminiferous tubules has remained unclear to date. Therefore, in this study, we investigated luminal flow and movements in the seminiferous tubules by three-dimensional analysis and in vivo imaging. Serial 5-μm-thick mouse testicular sections at 50-µm-intervals were prepared and stained by Periodic Acid-Schiff-hematoxylin. After three-dimensional reconstruction of the seminiferous tubules, the localization of the released spermatozoa and the stages observed in the sections were recorded in each reconstructed tubule. Luminal movements in the seminiferous tubules were observed by in vivo imaging using a fluorescent-reporter mouse and two-photon excitation microscopy system. Spermatozoa without contact to the seminiferous epithelium were not accumulated toward the rete testis. Additionally, such spermatozoa were found on their way not only to the most proximal rete testis but also a more distant rete testis from any stage VIII seminiferous epithelia. In vivo imaging demonstrated that the direction of the flagella of spermatozoa attached to the seminiferous epithelium was repeatedly reversed. The epithelium at the inner curve of the seminiferous tubule was shaken more actively and had fewer spermatozoa attached compared with the epithelium at the outer curve. Our results hence suggest that the luminal flow in the seminiferous tubules is repeatedly reversed and that this physical force helps spermatozoa to be released from Sertoli cells.

 

     An official journal of

    Society for Reproduction and Fertility

 

Sept 2018 onwards Past Year Past 30 Days
Abstract Views 952 952 663
Full Text Views 41 41 30
PDF Downloads 43 43 36
  • Abe T, Kiyonari H, Shioi G, Inoue K-I, Nakao K, Aizawa S & Fujimori T 2011 Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging. Genesis 49 579590. (https://doi.org/10.1002/dvg.20753)

    • Search Google Scholar
    • Export Citation
  • Albrechtová J, Albrecht T, Ďureje L, Pallazola VA & Piálek J 2014 Sperm morphology in two house mouse subspecies: do wild-derived strains and wild mice tell the same story? PLoS ONE 9 e115669. (https://doi.org/10.1371/journal.pone.0115669)

    • Search Google Scholar
    • Export Citation
  • Anton E 1979 Early ultrastructural changes in the rat testis after ductuli efferentes ligation. Fertility and Sterility 31 187194. (https://doi.org/10.1016/s0015-0282(1643821-5)

    • Search Google Scholar
    • Export Citation
  • Bucci LR & Meistrich ML 1987 Effects of busulfan on murine spermatogenesis: cytotoxicity, sterility, sperm abnormalities, and dominant lethal mutations. Mutation Research 176 259268. (https://doi.org/10.1016/0027-5107(8790057-1)

    • Search Google Scholar
    • Export Citation
  • Cerilli LA, Kuang W & & Rogers D 2010 A practical approach to testicular biopsy interpretation for male infertility. Archives of Pathology &. Laboratory Medicine 134 11971204. (https://doi.org/10.5858/2009-0379-RA.1)

    • Search Google Scholar
    • Export Citation
  • Clulow J, Jones RC & Hansen LA 1994 Micropuncture and cannulation studies of fluid composition and transport in the ductuli efferentes testis of the rat: comparisons with the homologous metanephric proximal tubule. Experimental Physiology 79 915928. (https://doi.org/10.1113/expphysiol.1994.sp003817)

    • Search Google Scholar
    • Export Citation
  • Fleck D, Kenzler L, Mundt N, Strauch M, Uesaka N, Moosmann R, Bruentgens F, Missel A, Mayerhofer A & Merhof D et al.2021 ATP activation of peritubular cells drives testicular sperm transport. eLife 10 e62885. (https://doi.org/10.7554/eLife.62885)

    • Search Google Scholar
    • Export Citation
  • Hess RA & de Franca LR 2009 Spermatogenesis and Cycle of the Seminiferous Epithelium In: Molecular Mechanisms in Spermatogenesis: Advances in Experimental Medicine and Biology. Cheng CY (Ed) New York, NY: Springer. (https://doi.org/10.1007/978-0-387-09597-4_1)

    • Search Google Scholar
    • Export Citation
  • Hinton BT & Keefer DA 1983 Evidence for protein absorption from the lumen of the seminiferous tubule and rete of the rat testis. Cell and Tissue Research 230 367375. (https://doi.org/10.1007/BF00213810)

    • Search Google Scholar
    • Export Citation
  • Holstein AF, Schulze W & Davidoff M 2003 Understanding spermatogenesis is a prerequisite for treatment. Reproductive Biology and Endocrinology 1 107. (https://doi.org/10.1186/1477-7827-1-107)

    • Search Google Scholar
    • Export Citation
  • Inaba K 2011 Sperm flagella: comparative and phylogenetic perspectives of protein components. Molecular Human Reproduction 17 524538. (https://doi.org/10.1093/molehr/gar034)

    • Search Google Scholar
    • Export Citation
  • Kiyozumi D, Noda T, Yamaguchi R, Tobita T, Matsumura T, Shimada K, Kodani M, Kohda T, Fujihara Y & Ozawa M et al.2020 NELL2-mediated lumicrine signaling through OVCH2 is required for male fertility. Science 368 11321135. (https://doi.org/10.1126/science.aay5134)

    • Search Google Scholar
    • Export Citation
  • Lee JSH, Panorchan P, Hale CM, Khatau SB, Kole TP, Tseng Y & Wirtz D 2006 Ballistic intracellular nanorheology reveals ROCK-hard cytoplasmic stiffening response to fluid flow. Journal of Cell Science 119 17601768. (https://doi.org/10.1242/jcs.02899)

    • Search Google Scholar
    • Export Citation
  • Lie PPY, Mruk DD, Lee WM & Cheng CY 2010 Cytoskeletal dynamics and spermatogenesis. Philosophical Transactions of the Royal Society of London: Series B, Biological Sciences 365 15811592. (https://doi.org/10.1098/rstb.2009.0261)

    • Search Google Scholar
    • Export Citation
  • Losinno AD, Sorrivas V, Ezquer M, Ezquer F, López LA & Morales A 2016 Changes of myoid and endothelial cells in the peritubular wall during contraction of the seminiferous tubule. Cell and Tissue Research 365 425435. (https://doi.org/10.1007/s00441-016-2386-x)

    • Search Google Scholar
    • Export Citation
  • Maekawa M, Kamimura K & Nagano T 1996 Peritubular myoid cells in the testis: their structure and function. Archives of Histology and Cytology 59 113. (https://doi.org/10.1679/aohc.59.1)

    • Search Google Scholar
    • Export Citation
  • Nakata H & Iseki S 2019 Three-dimensional structure of efferent and epididymal ducts in mice. Journal of Anatomy 235 271280. (https://doi.org/10.1111/joa.13006)

    • Search Google Scholar
    • Export Citation
  • Nakata H, Wakayama T, Takai Y & Iseki S 2015a Quantitative analysis of the cellular composition in seminiferous tubules in normal and genetically modified infertile mice. Journal of Histochemistry and Cytochemistry 63 99113. (https://doi.org/10.1369/0022155414562045)

    • Search Google Scholar
    • Export Citation
  • Nakata H, Wakayama T, Sonomura T, Honma S, Hatta T & Iseki S 2015b Three-dimensional structure of seminiferous tubules in the adult mouse. Journal of Anatomy 227 686694. (https://doi.org/10.1111/joa.12375)

    • Search Google Scholar
    • Export Citation
  • Nakata H, Sonomura T & Iseki S 2017 Three-dimensional analysis of seminiferous tubules and spermatogenic waves in mice. Reproduction 154 569579. (https://doi.org/10.1530/REP-17-0391)

    • Search Google Scholar
    • Export Citation
  • Nakata H, Nakano T, Iseki S & Mizokami A 2020 Three-dimensional analysis of busulfan-induced spermatogenesis disorder in mice. Frontiers in Cell and Developmental Biology 8 609278. (https://doi.org/10.3389/fcell.2020.609278)

    • Search Google Scholar
    • Export Citation
  • O’Donnell L, Nicholls PK, O’Bryan MK, McLachlan RI & Stanton PG 2011 Spermiation: the process of sperm release. Spermatogenesis 1 1435. (https://doi.org/10.4161/spmg.1.1.14525)

    • Search Google Scholar
    • Export Citation
  • Ohyama T & Groves AK 2004 Generation of Pax2-Cre mice by modification of a Pax2 bacterial artificial chromosome. Genesis 38 195199. (https://doi.org/10.1002/gene.20017)

    • Search Google Scholar
    • Export Citation
  • Russell LD, Ettlin RA, Hikim APS & Clegg ED 1990 Histological and Histopathological Evaluation of the Testis. Clearwater: Cache River Press.

  • Soler C, Yeung CH & Cooper TG 1994 Development of sperm motility patterns in the murine epididymis. International Journal of Andrology 17 271278. (https://doi.org/10.1111/j.1365-2605.1994.tb01253.x)

    • Search Google Scholar
    • Export Citation
  • Suto J-I 2008 Genetic dissection of testis weight in a mouse strain having an extremely large testis: major testis weight determinants are autosomal rather than Y-linked on the basis of comprehensive analyses in Y-chromosome consomic strains. PNAS 84 393406. (https://doi.org/10.2183/pjab.84.393)

    • Search Google Scholar
    • Export Citation
  • Uchida A, Sakib S, Labit E, Abbasi S, Scott RW, Underhill TM, Biernaskie J & Dobrinski I 2020 Development and function of smooth muscle cells is modulated by Hic1 in mouse testis. Development 147 dev185884. (https://doi.org/10.1242/dev.185884)

    • Search Google Scholar
    • Export Citation
  • Wen Q, Tang EI, Lui WY, Lee WM, Wong CKC, Silvestrini B & Cheng CY 2018 Dynein 1 supports spermatid transport and spermiation during spermatogenesis in the rat testis. American Journal of Physiology: Endocrinology and Metabolism 315 E924E948. (https://doi.org/10.1152/ajpendo.00114.2018)

    • Search Google Scholar
    • Export Citation
  • Yan Cheng C & Mruk DD 2015 Biochemistry of Sertoli cell/germ cell junctions, germ cell transport, and spermiation in the seminiferous epithelium. In Sertoli Cell Biology, 2 nd ed., vol. 12, pp. 333383. Ed Griswold MD Oxford: Academic Press. (https://doi.org/10.1016/B978-0-12-417047-6.00012-0)

    • Search Google Scholar
    • Export Citation
  • Yuan S, Liu Y, Peng H, Tang C, Hennig GW, Wang Z, Wang L, Yu T, Klukovich R & Zhang Y et al.2019 Motile cilia of the male reproductive system require miR-34/miR-449 for development and function to generate luminal turbulence. PNAS 116 35843593. (https://doi.org/10.1073/pnas.1817018116)

    • Search Google Scholar
    • Export Citation