Desert hedgehog mediates the proliferation of medaka spermatogonia through Smoothened signaling

in Reproduction
Authors:
Changle ZhaoKey Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China

Search for other papers by Changle Zhao in
Current site
Google Scholar
PubMedClose
,
Zeming ZhangKey Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China

Search for other papers by Zeming Zhang in
Current site
Google Scholar
PubMedClose
,
Ximei QuKey Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China

Search for other papers by Ximei Qu in
Current site
Google Scholar
PubMedClose
,
Xiaoming BaiKey Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China

Search for other papers by Xiaoming Bai in
Current site
Google Scholar
PubMedClose
,
Xingyong LiuKey Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China

Search for other papers by Xingyong Liu in
Current site
Google Scholar
PubMedClose
,
Wenjing TaoKey Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China

Search for other papers by Wenjing Tao in
Current site
Google Scholar
PubMedClose
,
Linyan ZhouKey Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China

Search for other papers by Linyan Zhou in
Current site
Google Scholar
PubMedClose
,
Deshou WangKey Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China

Search for other papers by Deshou Wang in
Current site
Google Scholar
PubMedClose
, and
Jing WeiKey Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, China

Search for other papers by Jing Wei in
Current site
Google Scholar
PubMedClose
https://orcid.org/0000-0003-1062-6691
View More View Less

Correspondence should be addressed to D Wang or J Wei; Email: wdeshou@swu.edu.cn or lalsos@swu.edu.cn
DOI:
https://doi.org/10.1530/REP-21-0468
Volume/Issue:
Volume 163: Issue 4
Page Range:
209–218
Article Type:
Research Article
Online Publication Date:
24 Feb 2022
Copyright:
© Society for Reproduction and Fertility 2022
Restricted access

USD  $0.01
USD  $0.01

USD  $0.01
USD  $0.01

USD  $0.01
USD  $0.01

USD  $0.01
USD  $0.01

USD  $0.01
USD  $0.01

USD  $0.01
USD  $0.01

USD  $0.01
USD  $0.01

USD  $1.00
USD  $1.00

Desert hedgehog (DHH) signaling has been reported to be involved in spermatogenesis and the self-renewal of spermatogonial stem cells (SSCs). However, the role of DHH in proliferation of spermatogonia including SSCs remains to be elucidated. Here, we report that Dhh from medaka (Oryizas latipes) (named as OlDhh) could directly mediate the proliferation of spermatogonia via Smoothened (Smo) signaling. Oldhh is 1362 bp in length and encodes 453 amino acid (aa) residues with more than 50% identity with the homologs in other species. It has expression predominantly restricted to testis. The soluble and tag-free 176-aa mature OlDhh (named as mOlDhh) were successfully obtained by fusing with the N-terminal tag of cleavable 6-histidine and small ubiquitin-related modifier and then removing the tag. Notably, mOlDhh significantly promoted the proliferation of SG3 (a spermatogonial stem cell line from medaka testis) in a dose-dependent manner and spermatogonia in testicular organ culture. Furthermore, the proliferation of SG3 in the presence of mOlDhh could be inhibited by Smo antagonist (cyclopamine) resulting in apoptosis. Additionally, mOlDhh significantly upregulated the expression of smo as well as the pluripotent-related genes bcl6b and sall4. These data suggest that Smo is an indispensable downstream component in the Dhh signaling pathway. In conclusion, our findings unambiguously demonstrate that Dhh could directly mediate the proliferation of spermatogonia through Smo signaling. This study provides new knowledge about the proliferation regulation of spermatogonia.

Reproduction is committed to supporting researchers in demonstrating the impact of their articles published in the journal.

The two types of article metrics we measure are (i) more traditional full-text views and pdf downloads, and (ii) Altmetric data, which shows the wider impact of articles in a range of non-traditional sources, such as social media.

More information is on the Reasons to publish page.

Sept 2018 onwards Past Year Past 30 Days
Full Text Views 74 30 4
PDF Downloads 77 24 3

 

  • Collapse
  • Expand
Print ISSN:
1470-1626
Online ISSN:
1741-7899

  • Bénazet JD, Bischofberger M, Tiecke E, Gonçalves A, Martin JF, Zuniga A, Naef F & Zeller R 2009 A self-regulatory system of interlinked signaling feedback loops controls mouse limb patterning. Science 323 1050–1053. (https://doi.org/10.1126/science.1168755)

    • Search Google Scholar
    • Export Citation

  • Bitgood MJ, Shen L & McMahon AP 1996 Sertoli cell signaling by desert hedgehog regulates the male germline. Current Biology 6 298–304. (https://doi.org/10.1016/s0960-9822(02)00480-3)

    • Search Google Scholar
    • Export Citation

  • Büller NV, Rosekrans SL, Westerlund J & van den Brink GR 2012 Hedgehog signaling and maintenance of homeostasis in the intestinal epithelium. Physiology 27 148–155. (https://doi.org/10.1152/physiol.00003.2012)

    • Search Google Scholar
    • Export Citation

  • Carpenter D, Stone DM, Brush J, Ryan A, Armanini M, Frantz G, Rosenthal A & de Sauvage FJ 1998 Characterization of two patched receptors for the vertebrate hedgehog protein family. PNAS 95 13630–13634. (https://doi.org/10.1073/pnas.95.23.13630)

    • Search Google Scholar
    • Export Citation

  • Chen MH, Li YJ, Kawakami T, Xu SM & Chuang PT 2004 Palmitoylation is required for the production of a soluble multimeric hedgehog protein complex and long-range signaling in vertebrates. Genes and Development 18 641–659. (https://doi.org/10.1101/gad.1185804)

    • Search Google Scholar
    • Export Citation

  • Chuang PT & McMahon AP 1999 Vertebrate hedgehog signalling modulated by induction of a hedgehog-binding protein. Nature 397 617–621. (https://doi.org/10.1038/17611)

    • Search Google Scholar
    • Export Citation

  • Clark AM, Garland KK & Russell LD 2000 Desert hedgehog (Dhh) gene is required in the mouse testis for formation of adult-type Leydig cells and normal development of peritubular cells and seminiferous tubules. Biology of Reproduction 63 1825–1838. (https://doi.org/10.1095/biolreprod63.6.1825)

    • Search Google Scholar
    • Export Citation

  • Dai S, Qi S, Wei X, Liu X, Li Y, Zhou X, Xiao H, Lu B, Wang D & Li M 2021 Germline sexual fate is determined by the antagonistic action of dmrt1 and foxl3/foxl2 in tilapia. Development 148 dev199380. (https://doi.org/10.1242/dev.199380)

    • Search Google Scholar
    • Export Citation

  • Delgado RN, Mansky B, Ahanger SH, Lu C, Andersen RE, Dou Y, Alvarez-Buylla A & Lim DA 2020 Maintenance of neural stem cell positional identity by mixed-lineage leukemia 1. Science 368 48–53. (https://doi.org/10.1126/science.aba5960)

    • Search Google Scholar
    • Export Citation

  • Fuccillo M, Joyner AL & Fishell G 2006 Morphogen to mitogen: the multiple roles of hedgehog signalling in vertebrate neural development. Nature Reviews: Neuroscience 7 772–783. (https://doi.org/10.1038/nrn1990)

    • Search Google Scholar
    • Export Citation

  • Gu Y, Wang Y, He L, Zhang J, Zhu X, Liu N, Wang J, Lu T, He L & Tian Y et al.2021 Circular RNA circIPO11 drives self-renewal of liver cancer initiating cells via hedgehog signaling. Molecular Cancer 20 132. (https://doi.org/10.1186/s12943-021-01435-2)

    • Search Google Scholar
    • Export Citation

  • Hobbs RM, Fagoonee S, Papa A, Webster K, Altruda F, Nishinakamura R, Chai L & Pandolfi PP 2012 Functional antagonism between Sall4 and Plzf defines germline progenitors. Cell Stem Cell 10 284–298. (https://doi.org/10.1016/j.stem.2012.02.004)

    • Search Google Scholar
    • Export Citation

  • Hong Y, Liu T, Zhao H, Xu H, Wang W, Liu R, Chen T, Deng J & Gui J 2004 Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro. PNAS 101 8011–8016. (https://doi.org/10.1073/pnas.0308668101)

    • Search Google Scholar
    • Export Citation

  • Hopyan S, Gokgoz N, Poon R, Gensure RC, Yu C, Cole WG, Bell RS, Jüppner H, Andrulis IL & Wunder JS et al.2002 A mutant PTH/PTHrP type I receptor in enchondromatosis. Nature Genetics 30 306–310. (https://doi.org/10.1038/ng844)

    • Search Google Scholar
    • Export Citation

  • Huycke TR, Miller BM, Gill HK, Nerurkar NL, Sprinzak D, Mahadevan L & Tabin CJ 2019 Genetic and mechanical regulation of intestinal smooth muscle development. Cell 179 90 .e21–105.e21. (https://doi.org/10.1016/j.cell.2019.08.041)

    • Search Google Scholar
    • Export Citation

  • Inaba M, Yamashita YM & Buszczak M 2016 Keeping stem cells under control: new insights into the mechanisms that limit niche-stem cell signaling within the reproductive system. Molecular Reproduction and Development 83 675–683. (https://doi.org/10.1002/mrd.22682)

    • Search Google Scholar
    • Export Citation

  • Ingham PW & Placzek M 2006 Orchestrating ontogenesis: variations on a theme by sonic hedgehog. Nature Reviews: Genetics 7 841–850. (https://doi.org/10.1038/nrg1969)

    • Search Google Scholar
    • Export Citation

  • Ishikura Y, Ohta H, Sato T, Murase Y, Yabuta Y, Kojima Y, Yamashiro C, Nakamura T, Yamamoto T & Ogawa T et al.2021 In vitro reconstitution of the whole male germ-cell development from mouse pluripotent stem cells. Cell Stem Cell 28 2167 .e9–2179.e9. (https://doi.org/10.1016/j.stem.2021.08.005)

    • Search Google Scholar
    • Export Citation

  • Jiang D, Chen J, Fan Z, Tan D, Zhao J, Shi H, Liu Z, Tao W, Li M & Wang D 2017 CRISPR/Cas9-induced disruption of wt1a and wt1b reveals their different roles in kidney and gonad development in Nile tilapia. Developmental Biology 428 63–73. (https://doi.org/10.1016/j.ydbio.2017.05.017)

    • Search Google Scholar
    • Export Citation

  • Kalderon D 2000 Transducing the hedgehog signal. Cell 103 371–374. (https://doi.org/10.1016/s0092-8674(00)00129-x)

  • Li S, Wang M, Chen Y, Wang W, Wu J, Yu C, Zheng Y & Pan Z 2017 Role of the hedgehog signaling pathway in regulating the behavior of germline stem cells. Stem Cells International 2017 5714608. (https://doi.org/10.1155/2017/5714608)

    • Search Google Scholar
    • Export Citation

  • Lovelace DL, Gao Z, Mutoji K, Song YC, Ruan J & Hermann BP 2016 The regulatory repertoire of PLZF and SALL4 in undifferentiated spermatogonia. Development 143 1893–1906. (https://doi.org/10.1242/dev.132761)

    • Search Google Scholar
    • Export Citation

  • Mäkelä JA, Saario V, Bourguiba-Hachemi S, Nurmio M, Jahnukainen K, Parvinen M & Toppari J 2011 Hedgehog signalling promotes germ cell survival in the rat testis. Reproduction 142 711–721. (https://doi.org/10.1530/REP-11-0110)

    • Search Google Scholar
    • Export Citation

  • Mann RK & Beachy PA 2004 Novel lipid modifications of secreted protein signals. Annual Review of Biochemistry 73 891–923. (https://doi.org/10.1146/annurev.biochem.73.011303.073933)

    • Search Google Scholar
    • Export Citation

  • Martinez-Chinchilla P & Riobo NA 2008 Purification and bioassay of hedgehog ligands for the study of cell death and survival. Methods in Enzymology 446 189–204. (https://doi.org/10.1016/S0076-6879(08)01611-X)

    • Search Google Scholar
    • Export Citation

  • Matunis EL, Stine RR & de Cuevas M 2012 Recent advances in Drosophila male germline stem cell biology. Spermatogenesis 2 137–144. (https://doi.org/10.4161/spmg.21763)

    • Search Google Scholar
    • Export Citation

  • Meng X, Lindahl M, Hyvönen ME, Parvinen M, de Rooij DG, Hess MW, Raatikainen-Ahokas A, Sainio K, Rauvala H & Lakso M et al.2000 Regulation of cell fate decision of undifferentiated spermatogonia by GDNF. Science 287 1489–1493. (https://doi.org/10.1126/science.287.5457.1489)

    • Search Google Scholar
    • Export Citation

  • Michel M, Kupinski AP, Raabe I & Bökel C 2012 Hh signalling is essential for somatic stem cell maintenance in the Drosophila testis niche. Development 139 2663–2669. (https://doi.org/10.1242/dev.075242)

    • Search Google Scholar
    • Export Citation

  • Morales CR, Fox A, El-Alfy M, Ni X & Argraves WS 2009 Expression of patched-1 and smoothened in testicular meiotic and post-meiotic cells. Microscopy Research and Technique 72 809–815. (https://doi.org/10.1002/jemt.20733)

    • Search Google Scholar
    • Export Citation

  • Naeemi S, Eidi A, Khanbabaee R, Sadri-Ardekani H & Kajbafzadeh AM 2021 Differentiation and proliferation of spermatogonial stem cells using a three-dimensional decellularized testicular scaffold: a new method to study the testicular microenvironment in vitro. International Urology and Nephrology 53 1543–1550. (https://doi.org/10.1007/s11255-021-02877-9)

    • Search Google Scholar
    • Export Citation

  • Nüsslein-Volhard C & Wieschaus E 1980 Mutations affecting segment number and polarity in Drosophila. Nature 287 795–801. (https://doi.org/10.1038/287795a0)

    • Search Google Scholar
    • Export Citation

  • Oatley JM & Brinster RL 2012 The germline stem cell niche unit in mammalian testes. Physiological Reviews 92 577–595. (https://doi.org/10.1152/physrev.00025.2011)

    • Search Google Scholar
    • Export Citation

  • Oatley JM, Avarbock MR, Telaranta AI, Fearon DT & Brinster RL 2006 Identifying genes important for spermatogonial stem cell self-renewal and survival. PNAS 103 9524–9529. (https://doi.org/10.1073/pnas.0603332103)

    • Search Google Scholar
    • Export Citation

  • Pak E & Segal RA 2016 Hedgehog signal transduction: key players, oncogenic drivers, and cancer therapy. Developmental Cell 38 333–344. (https://doi.org/10.1016/j.devcel.2016.07.026)

    • Search Google Scholar
    • Export Citation

  • Paredes I, Vieira JR, Shah B, Ramunno CF, Dyckow J, Adler H, Richter M, Schermann G, Giannakouri E & Schirmer L et al.2021 Oligodendrocyte precursor cell specification is regulated by bidirectional neural progenitor-endothelial cell crosstalk. Nature Neuroscience 24 478–488. (https://doi.org/10.1038/s41593-020-00788-z)

    • Search Google Scholar
    • Export Citation

  • Porter JA, von Kessler DP, Ekker SC, Young KE, Lee JJ, Moses K & Beachy PA 1995 The product of hedgehog autoproteolytic cleavage active in local and long-range signalling. Nature 374 363–366. (https://doi.org/10.1038/374363a0)

    • Search Google Scholar
    • Export Citation

  • Sahin Z, Szczepny A, McLaughlin EA, Meistrich ML, Zhou W, Ustunel I & Loveland KL 2014 Dynamic hedgehog signalling pathway activity in germline stem cells. Andrology 2 267–274. (https://doi.org/10.1111/j.2047-2927.2014.00187.x)

    • Search Google Scholar
    • Export Citation

  • Szczepny A, Hime GR & Loveland KL 2006 Expression of hedgehog signalling components in adult mouse testis. Developmental Dynamics 235 3063–3070. (https://doi.org/10.1002/dvdy.20931)

    • Search Google Scholar
    • Export Citation

  • Taipale J, Chen JK, Cooper MK, Wang B, Mann RK, Milenkovic L, Scott MP & Beachy PA 2000 Effects of oncogenic mutations in smoothened and patched can be reversed by cyclopamine. Nature 406 1005–1009. (https://doi.org/10.1038/35023008)

    • Search Google Scholar
    • Export Citation

  • Tan K & Wilkinson MF 2019 Human spermatogonial stem cells scrutinized under the single-cell magnifying glass. Cell Stem Cell 24 201–203. (https://doi.org/10.1016/j.stem.2019.01.010)

    • Search Google Scholar
    • Export Citation

  • Umehara F, Tate G, Itoh K, Yamaguchi N, Douchi T, Mitsuya T & Osame M 2000 A novel mutation of desert hedgehog in a patient with 46,XY partial gonadal dysgenesis accompanied by minifascicular neuropathy. American Journal of Human Genetics 67 1302–1305. (https://doi.org/10.1016/S0002-9297(07)62958-9)

    • Search Google Scholar
    • Export Citation

  • Vercauteren Drubbel A, Pirard S, Kin S, Dassy B, Lefort A, Libert F, Nomura S & Beck B 2021 Reactivation of the hedgehog pathway in esophageal progenitors turns on an embryonic-like program to initiate columnar metaplasia. Cell Stem Cell 28 1411 .e7–1427.e7. (https://doi.org/10.1016/j.stem.2021.03.019)

    • Search Google Scholar
    • Export Citation

  • Vortkamp A, Lee K, Lanske B, Segre GV, Kronenberg HM & Tabin CJ 1996 Regulation of rate of cartilage differentiation by Indian hedgehog and PTH-related protein. Science 273 613–622. (https://doi.org/10.1126/science.273.5275.613)

    • Search Google Scholar
    • Export Citation

  • Wang D, Manali D, Wang T, Bhat N, Hong N, Li Z, Wang L, Yan Y, Liu R & Hong Y 2011 Identification of pluripotency genes in the fish medaka. International Journal of Biological Sciences 7 440–451. (https://doi.org/10.7150/ijbs.7.440)

    • Search Google Scholar
    • Export Citation

  • Wei J, Liu L, Fan Z, Hong Y, Zhao Y, Zhou L & Wang D 2017 Identification, prokaryote expression of medaka gdnfa/b and their biological activity in a spermatogonial cell line. Stem Cells and Development 26 197–205. (https://doi.org/10.1089/scd.2016.0248)

    • Search Google Scholar
    • Export Citation

  • Werner R, Merz H, Birnbaum W, Marshall L, Schröder T, Reiz B, Kavran JM, Bäumer T, Capetian P & Hiort O 2015 46,XY gonadal dysgenesis due to a homozygous mutation in desert hedgehog (DHH) identified by exome sequencing. Journal of Clinical Endocrinology and Metabolism 100 E1022–E1029. (https://doi.org/10.1210/jc.2015-1314)

    • Search Google Scholar
    • Export Citation

  • Wu X, Goodyear SM, Tobias JW, Avarbock MR & Brinster RL 2011 Spermatogonial stem cell self-renewal requires ETV5-mediated downstream activation of brachyury in mice. Biology of Reproduction 85 1114–1123. (https://doi.org/10.1095/biolreprod.111.091793)

    • Search Google Scholar
    • Export Citation

  • Xie X, Nóbrega R & Pšenička M 2020 Spermatogonial stem cells in fish: characterization, isolation, enrichment, and recent advances of in vitro culture systems. Biomolecules 10 644. (https://doi.org/10.3390/biom10040644)

    • Search Google Scholar
    • Export Citation

  • Xu H, Lim M, Dwarakanath M & Hong Y 2014 Vasa identifies germ cells and critical stages of oogenesis in the Asian seabass. International Journal of Biological Sciences 10 225–235. (https://doi.org/10.7150/ijbs.6797)

    • Search Google Scholar
    • Export Citation

  • Yi M, Hong N & Hong Y 2009 Generation of medaka fish haploid embryonic stem cells. Science 326 430–433. (https://doi.org/10.1126/science.1175151)

    • Search Google Scholar
    • Export Citation

  • Yoshida S 2008 Spermatogenic stem cell system in the mouse testis. Cold Spring Harbor Symposia on Quantitative Biology 73 25–32. (https://doi.org/10.1101/sqb.2008.73.046)

    • Search Google Scholar
    • Export Citation

  • Yoshida S, Sukeno M & Nabeshima Y 2007 A vasculature-associated niche for undifferentiated spermatogonia in the mouse testis. Science 317 1722–1726. (https://doi.org/10.1126/science.1144885)

    • Search Google Scholar
    • Export Citation

  • Zhang Z, Lv X, Jiang J, Zhang L & Zhao Y 2013a Dual roles of Hh signaling in the regulation of somatic stem cell self-renewal and germline stem cell maintenance in Drosophila testis. Cell Research 23 573–576. (https://doi.org/10.1038/cr.2013.29)

    • Search Google Scholar
    • Export Citation

  • Zhang Z, Pan C & Zhao Y 2013b Hedgehog in the Drosophila testis niche: what does it do there? Protein and Cell 4 650–655. (https://doi.org/10.1007/s13238-013-3040-y)

    • Search Google Scholar
    • Export Citation

  • Zhang X, Dong S & Xu F 2018 Structural and druggability landscape of frizzled G protein-coupled receptors. Trends in Biochemical Sciences 43 1033–1046. (https://doi.org/10.1016/j.tibs.2018.09.002)

    • Search Google Scholar
    • Export Citation

  • Zhao Y, Yang Z, Wang Y, Luo Y, Da F, Tao W, Zhou L, Wang D & Wei J 2018 Both Gfrα1a and Gfrα1b are involved in the self-renewal and maintenance of spermatogonial stem cells in medaka. Stem Cells and Development 27 1658–1670. (https://doi.org/10.1089/scd.2018.0177)

    • Search Google Scholar
    • Export Citation