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Biol Open
2016 Sep 15;59:1275-82. doi: 10.1242/bio.019265.
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Identification and characterization of Xenopus tropicalis common progenitors of Sertoli and peritubular myoid cell lineages.
Tlapakova T
,
Nguyen TM
,
Vegrichtova M
,
Sidova M
,
Strnadova K
,
Blahova M
,
Krylov V
.
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The origin of somatic cell lineages during testicular development is controversial in mammals. Employing basal amphibian tetrapod Xenopus tropicalis we established a cell culture derived from testes of juvenile male. Expression analysis showed transcription of some pluripotency genes and Sertoli cell, peritubular myoid cell and mesenchymal cell markers. Transcription of germline-specific genes was downregulated. Immunocytochemistry revealed that a majority of cells express vimentin and co-express Sox9 and smooth muscle α-actin (Sma), indicating the existence of a common progenitor of Sertoli and peritubular myoid cell lineages. Microinjection of transgenic, red fluorescent protein (RFP)-positive somatic testicular cells into the peritoneal cavity of X. tropicalis tadpoles resulted in cell deposits in heart, pronephros and intestine, and later in a strong proliferation and formation of cell-to-cell net growing through the tadpole body. Immunohistochemistry analysis of transplanted tadpoles showed a strong expression of vimentin in RFP-positive cells. No co-localization of Sox9 and Sma signals was observed during the first three weeks indicating their dedifferentiation to migratory-active mesenchymal cells recently described in human testicular biopsies.
Fig. 1. In vitro characterization of X. tropicalis cell culture. (A,B) Testicular somatic cell culture in morphology of adherent feeder layer (XtTSC) (A) and after long-term cultivation which enables the forming of colonies (XtTSCc) (B). (C) X. tropicalis transgenic XtTSC expressing Katushka RFP under CAG promotor (XtTSC-RFP). (D) Transgenic Katushka RFP expressing XtTSC in colonies (XtTSCc-RFP). (E,F) Structure of in vitro testicular cell colony visualized by TEM. In the colony the cells are placed in an extensive amount of extracellular matrix with two or three tight layers of XtTSCs surrounding the colony at the edge (E). Both XtTSC and XtTSCc are present in the centre of the colony (F). The XtTSCc are clearly several times smaller than the XtTSC. Red arrowheads, XtTSC; blue arrowheads, XtTSCc. (G) X. tropicalis cell culture proliferation during long-term cultivation in medium with or without recombinant mouse LIF. (H) Representative graph of FACS sorting after nucleofection. Only ∼15% of living transgenic cells with the highest intensity of fluorescent signal were sorted (blue area). Scale bars A,C:100 μm; scale bars B,D: 200 μm; scale bars E,F: 10 μm.
Fig. 2. Expression analysis of X. tropicalis testicular culture. (A) RT-PCR analysis of X. tropicalis testis, XtTSC, XtTSCc, XtTSC-RFP and XtTSCc-RFP. (B) Hierarchical clustering presented as a qPCR heatmaps of germ cell markers and selected testis associated markers. A scale of colours indicates level of expression (the highest expression is shown by bright red, whereas the lowest expression is shown by bright green). Similarity between cell types/genes is indicated by the height at which the dendrograms are joined. The RNA spike represents a highly stable transcript across the cell types and odc1 represents housekeeping gene.
Fig. 3. Immunocytochemistry of X. tropicalis testicular cell culture expressing Katushka RFP. Positive staining for Sma (marker of peritubular myoid cells, green) (A) and vimentin (marker of mesenchymal cells and peritubular myoid cells, green) (B). Nuclei were counterstained with DAPI (blue). (C-E) Double staining with Sox9 (marker of Sertoli cells, green) (C) and Sma (blue) (D) antibodies. (E) Merge of C and D. Yellow arrowhead, cell expressing both antigens; red arrowheads, cells expressing only Sox9 or only Sma. (F-H) Double staining with Sox9 (green) (F) and vimentin (blue) (G) antibodies. (H) Merge of F and G. Scale bars: 50 μm.
Fig. 4. Immunohistochemistry of agarose embedded testicular sections from X. tropicalis and mouse adult males. (A,C) Double staining with Sox9 (red) and Sma (green) antibodies. White arrowheads indicate potential common precursor cells for Sertoli and PTM cell lineages expressing both antigens in X. tropicalis (A) and mouse (C) samples. Insets show a detailed view of structures marked with white arrowheads in underlying figures. (B,D) Staining with vimentin (green) antibody on X. tropicalis (B) and mouse (D) samples. Nuclei were counterstained with DAPI (blue). Scale bars: 40 μm.
Fig. 5. Migration potential of X. tropicalis testicular somatic cells after allogeneic transplantation into peritoneal cavity of tadpoles in stage 41. (A) Scheme of XtTSCc-RFP preparation prior to transplantation employing isolation of cell colonies using 40 μm sieve and subsequent single cell dissociation with papain. 500 cells were microinjected dorsally into peritoneal cavity. (B-G) Observation of RFP-positive cells in transplanted tadpoles under stereo microscopy. (B) Cell deposit in pronephros and tail 1 day after microinjection. (C,D) Cell deposits in pronephros 13 days after microinjection. (E) Migration of RFP-positive cells into heart atrium 15 days after transplantation. (F,G) Cell-to-cell net growing through the tadpole body observable 30 days after microinjection. (H) Immunohistochemistry of agarose-embedded sections of transplanted tadpoles using antibodies against Sox9, Sma and vimentin (green) and Katushka RFP (red) 0, 1 and 30 days after transplantation. RFP-positive cells were stained with vimentin antibody but not Sox9 or Sma even 2 h after transplantation. 30 days after microinjection, few cells start to express Sox9 indicating potential redifferentiation into Sertoli cells or chondrocytes where Sox9 is considered as a specific marker. The first three figures in each line were taken under fluorescence microscopy. The figures on the right side were taken under fluorescence stereomicroscopy. Nuclei were counterstained with DAPI (blue). Scale bars B-G, H (0 and 1 day after transplantation), 300 μm; scale bars in H (30 days after transplantation, staining with Sox9 and Sma antibodies), 900 μm; scale bars in H (30 days after transplantation, staining with vimentin antibody), 200 μm.
Abe,
Identification, cDNA cloning, and mRNA localization of a zebrafish ortholog of leukemia inhibitory factor.
2007, Pubmed
Abe,
Identification, cDNA cloning, and mRNA localization of a zebrafish ortholog of leukemia inhibitory factor.
2007,
Pubmed
Baker,
Expression of 3beta-hydroxysteroid dehydrogenase type I and type VI isoforms in the mouse testis during development.
1999,
Pubmed
Berndtson,
Changing relationships between testis size, Sertoli cell number and spermatogenesis in Sprague-Dawley rats.
1990,
Pubmed
Bott,
Vascular endothelial growth factor and kinase domain region receptor are involved in both seminiferous cord formation and vascular development during testis morphogenesis in the rat.
2006,
Pubmed
Campagnolo,
Mesenchymal cell precursors of peritubular smooth muscle cells of the mouse testis can be identified by the presence of the p75 neurotrophin receptor.
2001,
Pubmed
Chikhovskaya,
Human testis-derived embryonic stem cell-like cells are not pluripotent, but possess potential of mesenchymal progenitors.
2012,
Pubmed
Chikhovskaya,
Mesenchymal origin of multipotent human testis-derived stem cells in human testicular cell cultures.
2014,
Pubmed
Chowdhury,
Soft substrates promote homogeneous self-renewal of embryonic stem cells via downregulating cell-matrix tractions.
2010,
Pubmed
Combes,
Endothelial cell migration directs testis cord formation.
2009,
Pubmed
Conrad,
Generation of pluripotent stem cells from adult human testis.
2008,
Pubmed
Cool,
Testis formation in the fetal mouse: dynamic and complex de novo tubulogenesis.
2012,
Pubmed
Cornelissen,
Organ-specific migration of mesenchymal stromal cells: Who, when, where and why?
2015,
Pubmed
De Miguel,
Leukemia inhibitory factor and ciliary neurotropic factor promote the survival of Sertoli cells and gonocytes in coculture system.
1996,
Pubmed
de Rooij,
The spermatogonial stem cell niche.
2009,
Pubmed
Dufour,
Neonatal porcine Sertoli cells inhibit human natural antibody-mediated lysis.
2005,
Pubmed
El Jamil,
Sex-specific expression of SOX9 during gonadogenesis in the amphibian Xenopus tropicalis.
2008,
Pubmed
,
Xenbase
Flachsova,
Single blastomere expression profiling of Xenopus laevis embryos of 8 to 32-cells reveals developmental asymmetry.
2013,
Pubmed
,
Xenbase
Frankenberg,
The POU-er of gene nomenclature.
2014,
Pubmed
Fujiki,
Molecular cloning and characterisation of a carp (Cyprinus carpio) cytokine-like cDNA that shares sequence similarity with IL-6 subfamily cytokines CNTF, OSM and LIF.
2003,
Pubmed
Geach,
Developmental genetics in Xenopus tropicalis.
2011,
Pubmed
,
Xenbase
Golestaneh,
Pluripotent stem cells derived from adult human testes.
2009,
Pubmed
Guan,
Pluripotency of spermatogonial stem cells from adult mouse testis.
2006,
Pubmed
Hanington,
Interleukin-6 family cytokine M17 induces differentiation and nitric oxide response of goldfish (Carassius auratus L.) macrophages.
2007,
Pubmed
Hanington,
Analysis of leukemia inhibitory factor and leukemia inhibitory factor receptor in embryonic and adult zebrafish (Danio rerio).
2008,
Pubmed
Hellsten,
The genome of the Western clawed frog Xenopus tropicalis.
2010,
Pubmed
,
Xenbase
Horiuchi,
Maintenance of chicken embryonic stem cells in vitro.
2006,
Pubmed
Hylis,
Aquatic tetrasporoblastic microsporidia from caddis flies (Insecta, Trichoptera): characterisation, phylogeny and taxonomic reevaluation of the genera Episeptum Larsson, 1986, Pyrotheca Hesse, 1935 and Cougourdella Hesse, 1935.
2007,
Pubmed
Inoue,
Isolation and Characterization of Fetal Leydig Progenitor Cells of Male Mice.
2016,
Pubmed
Jenab,
Testicular leukemia inhibitory factor (LIF) and LIF receptor mediate phosphorylation of signal transducers and activators of transcription (STAT)-3 and STAT-1 and induce c-fos transcription and activator protein-1 activation in rat Sertoli but not germ cells.
1998,
Pubmed
Johnson,
Quantification of the human Sertoli cell population: its distribution, relation to germ cell numbers, and age-related decline.
1984,
Pubmed
Kanatsu-Shinohara,
Long-term proliferation in culture and germline transmission of mouse male germline stem cells.
2003,
Pubmed
Kanatsu-Shinohara,
Generation of pluripotent stem cells from neonatal mouse testis.
2004,
Pubmed
Kanatsu-Shinohara,
Pluripotency of a single spermatogonial stem cell in mice.
2008,
Pubmed
Karl,
Sertoli cells of the mouse testis originate from the coelomic epithelium.
1998,
Pubmed
Kim,
Role of the epithelial-mesenchymal transition and its effects on embryonic stem cells.
2014,
Pubmed
Ko,
Induction of pluripotency in adult unipotent germline stem cells.
2009,
Pubmed
Koopman,
Expression of a candidate sex-determining gene during mouse testis differentiation.
1990,
Pubmed
Kossack,
Isolation and characterization of pluripotent human spermatogonial stem cell-derived cells.
2009,
Pubmed
Mathieu,
LIF-dependent signaling: new pieces in the Lego.
2012,
Pubmed
Mizrak,
Embryonic stem cell-like cells derived from adult human testis.
2010,
Pubmed
Morrison,
Conserved roles for Oct4 homologues in maintaining multipotency during early vertebrate development.
2006,
Pubmed
,
Xenbase
Nef,
A molecular basis for estrogen-induced cryptorchidism.
2000,
Pubmed
Nishino,
Establishment of fetal gonad/mesonephros coculture system using EGFP transgenic mice.
2000,
Pubmed
O'Shaughnessy,
Leydig cell re-generation and expression of cell signaling molecules in the germ cell-free testis.
2008,
Pubmed
Piquet-Pellorce,
Leukemia inhibitory factor expression and regulation within the testis.
2000,
Pubmed
Shan,
Developmental changes in levels of luteinizing hormone receptor and androgen receptor in rat Leydig cells.
1992,
Pubmed
Shcherbo,
Bright far-red fluorescent protein for whole-body imaging.
2007,
Pubmed
,
Xenbase
Sidova,
Intracellular microRNA profiles form in the Xenopus laevis oocyte that may contribute to asymmetric cell division.
2015,
Pubmed
,
Xenbase
Silva,
Nanog is the gateway to the pluripotent ground state.
2009,
Pubmed
Skinner,
Cooperativity between Sertoli cells and testicular peritubular cells in the production and deposition of extracellular matrix components.
1985,
Pubmed
Stanley,
Stem Leydig cell differentiation: gene expression during development of the adult rat population of Leydig cells.
2011,
Pubmed
Tymowska,
Karyotype analysis of Xenopus tropicalis Gray, Pipidae.
1973,
Pubmed
,
Xenbase
Wells,
A genetic map of Xenopus tropicalis.
2011,
Pubmed
,
Xenbase
