Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Sci Rep
2019 Oct 10;91:14652. doi: 10.1038/s41598-019-51224-7.
Show Gene links
Show Anatomy links
Discovery of a receptor guanylate cyclase expressed in the sperm flagella of stony corals.
Zhang Y
,
Chiu YL
,
Chen CJ
,
Ho YY
,
Shinzato C
,
Shikina S
,
Chang CF
.
???displayArticle.abstract???
The receptor guanylate cyclases (rGCs) in animals serve as sensitive chemoreceptors to detect both chemical and environmental cues. In reproduction, rGCs were shown to be expressed on sperm and serve as receptors for egg-derived sperm-activating and sperm-attracting factors in some echinoderms and mammals. However, sperm-associated rGCs have only been identified in some deuterostomes thus far, and it remains unclear how widely rGCs are utilized in metazoan reproduction. To address this issue, this study investigated the existence and expression of rGCs, particularly asking if rGCs are involved in the reproduction of a basal metazoan, phylum Cnidaria, using the stony coral Euphyllia ancora. Six paralogous rGCs were identified from a transcriptome database of E. ancora, and one of the rGCs, GC-A, was shown to be specifically expressed in the testis. Immunohistochemical analyses demonstrated that E. ancora GC-A protein was expressed in the spermatocytes and spermatids and eventually congregated on the sperm flagella during spermatogenesis. These findings suggest that GC-A may be involved in the regulation of sperm activity and/or functions (e.g., fertilization) in corals. This study is the first to perform molecular characterization of rGCs in cnidarians and provides evidence for the possible involvement of rGCs in the reproduction of basal metazoans.
Figure 1. The tissue distribution of rGC transcripts in E. ancora polyp and localization of GC-A-expressing cells. (a) Schematic of the E. ancora polyp structure (modified from Shikina et al., ref.57). (b–g) Expression of GC-A, GC-B, GC-C, GC-D, GC-E, and GC-F mRNAs in the different parts of polyp tissues in male and female E. ancora. (h) Comparison of mRNA levels of the six rGCs in the testis of E. ancora. (i) Comparison of mRNA levels of GC-A in the testis containing different developmental stages of male germ cells. The samples investigated include tentacle (Ten), testis (Tes), mesenterial filament (Mf), and ovary (Ova). Data shown are the mean ± SE (n = 3 colonies) relative to the Ten group. Groups with different letters are significantly different (P < 0.05). Because of the low expression levels of GC-B (c) and GC-F (j) mRNA, the transcripts were only detected in 1 out of 3 colonies examined (1/3) or 2 out of 3 colonies examined (2/3). ND, not detected. Localization of GC-A-expressing cells in E. ancora testis (j–l). Sequential sections were stained with hematoxylin-eosin (j), hybridized to an antisense GC-A probe (k), or hybridized to a sense GC-A probe (l). A fine dotted line distinguishes the edge of spermary and testicular somatic tissue. The GC-A signal was detected mainly in secondary spermatocytes and faintly in spermatids. spm, spermary; tsc, testicular somatic cell; sc II, secondary spermatocyte; std, spermatid. Scale bar, 20 μm.
Figure 2. Evaluation of the anti-GC-A antibody specificity and GC-A expression in the developing gonads. The antibody was used for the analysis with or without preadsorption of its peptide antigen. (a) SDS-PAGE of protein extracts prepared from developing ovaries (Ova) and testes (Tes) of E. ancora that were collected in April (1 month before spawning). Markers with molecular sizes are shown. (b) Western blotting of the same protein extracts shown in (a), probed with an anti-GC-A antibody. An immunoreactive band at 155 kDa is shown. Note that the immunoreactivity was completely eliminated by the preadsorption of the antibody with the peptide antigens (+antigen). The amount of protein used was 10 µg for each tissue.
Figure 3. The expression of GC-A, Ac-α-Tu, and EaPiwi in male germ cells at different stages of spermatogenesis. (a,d,g,j) Expression profile of GC-A. (b,e,h,k) Expression profile of Ac-α-Tu. (c,f,i,l) Expression profile of EaPiwi. Sequential sections were used to analyse each stage of male germ cells. Scale bar, 20 μm.
Figure 4. The localization of GC-A expression in the sperm. (a) SDS-PAGE and western blotting of sperm protein. Molecular size markers are shown. An immunoreactive band at 155 kDa is shown. (b) Double immunofluorescence detection of acetylated-alpha-tubulin, Ac-α-Tu (upper, green) and GC-A (middle, red) in the flagella of sperm in a paraffin section of E. ancora testis. Ac-α-Tu and GC-A were colocalized in the sperm flagella (lower, merge). (c) Immunohistochemical analysis of a mature testis. The immunoreactivity was concentrated in the sperm flagella and almost eliminated by the preadsorption of the antibody with the peptide antigens. The amount of protein used was 10 µg for each tissue. Scale bar, 20 μm.
Figure 5. Presence of GC-A in the male germ cells of different stony corals as assessed by immunohistochemistry. (a) Sperm of Euphyllia glabrescens, (b) Spermatids of Echinophyllia echinoporoides, (c) Spermatogonia/primary spermatocytes of Acropora sp., (d) Spermatogonia/primary spermatocytes of Hydnophora sp., (e) Sperm of Pectinia lectuca, (f) Sperm of Pectinia peaonia, (g) Sperm of Favites pentagona, (h) Sperm of Goniastrea australiensis, (i) Sperm of Platygyra daedalea, (j) Sperm of Platygyra lamellina, (k) Sperm of Pachyseris speciosa, (l) Sperm of Goniopora sp. Scale bar, 20 μm.
Albalat,
Evolution by gene loss.
2016,
Pubmed
Anderson,
Atrial natriuretic peptide (ANP) as a stimulus of the human acrosome reaction and a component of ovarian follicular fluid: correlation of follicular ANP content with in vitro fertilization outcome.
1994,
Pubmed
Anderson,
Atrial natriuretic peptide: a chemoattractant of human spermatozoa by a guanylate cyclase-dependent pathway.
1995,
Pubmed
,
Echinobase
Arshad,
The multiple and enigmatic roles of guanylyl cyclase C in intestinal homeostasis.
2012,
Pubmed
Audagnotto,
Protein post-translational modifications: In silico prediction tools and molecular modeling.
2017,
Pubmed
Basu,
Intestinal cell proliferation and senescence are regulated by receptor guanylyl cyclase C and p21.
2014,
Pubmed
Bian,
Gradients of natriuretic peptide precursor A (NPPA) in oviduct and of natriuretic peptide receptor 1 (NPR1) in spermatozoon are involved in mouse sperm chemotaxis and fertilization.
2012,
Pubmed
Chapman,
The dynamic genome of Hydra.
2010,
Pubmed
Collins,
Phylogenetic context and Basal metazoan model systems.
2005,
Pubmed
Duan,
The roles of post-translational modifications in the context of protein interaction networks.
2015,
Pubmed
Eitel,
Comparative genomics and the nature of placozoan species.
2018,
Pubmed
El-Gehani,
Natriuretic peptides stimulate steroidogenesis in the fetal rat testis.
2001,
Pubmed
Fenrick,
Glycosylation is critical for natriuretic peptide receptor-B function.
1996,
Pubmed
Fitzpatrick,
Multiple lineage specific expansions within the guanylyl cyclase gene family.
2006,
Pubmed
Foster,
Mechanisms of regulation and functions of guanylyl cyclases.
1999,
Pubmed
Geister,
A novel loss-of-function mutation in Npr2 clarifies primary role in female reproduction and reveals a potential therapy for acromesomelic dysplasia, Maroteaux type.
2013,
Pubmed
Ghanekar,
Glycosylation of the receptor guanylate cyclase C: role in ligand binding and catalytic activity.
2004,
Pubmed
Healy,
Natriuretic peptide guanylyl cyclase receptors in the kidney of the Japanese eel, Anguilla japonica.
2005,
Pubmed
Kaupp,
Sperm chemotaxis in marine invertebrates--molecules and mechanisms.
2006,
Pubmed
Koller,
Proper glycosylation and phosphorylation of the type A natriuretic peptide receptor are required for hormone-stimulated guanylyl cyclase activity.
1993,
Pubmed
Kong,
Natriuretic peptide type C induces sperm attraction for fertilization in mouse.
2017,
Pubmed
Kortschak,
EST analysis of the cnidarian Acropora millepora reveals extensive gene loss and rapid sequence divergence in the model invertebrates.
2003,
Pubmed
Kuhn,
Molecular Physiology of Membrane Guanylyl Cyclase Receptors.
2016,
Pubmed
Kumar,
MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms.
2018,
Pubmed
Leclère,
The genome of the jellyfish Clytia hemisphaerica and the evolution of the cnidarian life-cycle.
2019,
Pubmed
Livak,
Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.
2001,
Pubmed
Maruyama,
Receptor Guanylyl Cyclases in Sensory Processing.
2016,
Pubmed
Nishigaki,
A 130-kDa membrane protein of sperm flagella is the receptor for asterosaps, sperm-activating peptides of starfish Asterias amurensis.
2000,
Pubmed
,
Echinobase
Pichlo,
High density and ligand affinity confer ultrasensitive signal detection by a guanylyl cyclase chemoreceptor.
2014,
Pubmed
,
Echinobase
Potter,
Guanylyl cyclase structure, function and regulation.
2011,
Pubmed
Potter,
Guanylyl cyclase-linked natriuretic peptide receptors: structure and regulation.
2001,
Pubmed
Putnam,
Sea anemone genome reveals ancestral eumetazoan gene repertoire and genomic organization.
2007,
Pubmed
Rappaport,
The Guanylate Cyclase C-cGMP Signaling Axis Opposes Intestinal Epithelial Injury and Neoplasia.
2018,
Pubmed
Revelli,
Guanylate cyclase activity and sperm function.
2002,
Pubmed
Riesgo,
The analysis of eight transcriptomes from all poriferan classes reveals surprising genetic complexity in sponges.
2014,
Pubmed
Roelofs,
Genes lost during evolution.
2001,
Pubmed
Ryan,
The genome of the ctenophore Mnemiopsis leidyi and its implications for cell type evolution.
2013,
Pubmed
Rätscho,
Expression profiles of three novel sensory guanylate cyclases and guanylate cyclase-activating proteins in the zebrafish retina.
2009,
Pubmed
Shikina,
Germ cell development in the scleractinian coral Euphyllia ancora (Cnidaria, Anthozoa).
2012,
Pubmed
Shikina,
Immunodetection of acetylated alpha-tubulin in stony corals: Evidence for the existence of flagella in coral male germ cells.
2017,
Pubmed
Shikina,
Molecular cloning and characterization of a steroidogenic enzyme, 17β-hydroxysteroid dehydrogenase type 14, from the stony coral Euphyllia ancora (Cnidaria, Anthozoa).
2016,
Pubmed
Shikina,
Yolk formation in a stony coral Euphyllia ancora (Cnidaria, Anthozoa): insight into the evolution of vitellogenesis in nonbilaterian animals.
2013,
Pubmed
Shinzato,
Using the Acropora digitifera genome to understand coral responses to environmental change.
2011,
Pubmed
Shuhaibar,
Intercellular signaling via cyclic GMP diffusion through gap junctions restarts meiosis in mouse ovarian follicles.
2015,
Pubmed
Technau,
Maintenance of ancestral complexity and non-metazoan genes in two basal cnidarians.
2005,
Pubmed
Walsh,
Protein posttranslational modifications: the chemistry of proteome diversifications.
2005,
Pubmed
Ward,
Phosphorylation of membrane-bound guanylate cyclase of sea urchin spermatozoa.
1986,
Pubmed
,
Echinobase
Xia,
Role of C-type natriuretic peptide in the function of normal human sperm.
2016,
Pubmed
Xia,
C-type natriuretic peptide regulates blood-testis barrier dynamics in adult rat testes.
2007,
Pubmed
Yang,
Characterization of a novel cell-surface protein expressed on human sperm.
2010,
Pubmed
Zamir,
Atrial natriuretic peptide attracts human spermatozoa in vitro.
1993,
Pubmed
Zhang,
Granulosa cell ligand NPPC and its receptor NPR2 maintain meiotic arrest in mouse oocytes.
2010,
Pubmed
Zhang,
Porcine natriuretic peptide type B (pNPPB) maintains mouse oocyte meiotic arrest via natriuretic peptide receptor 2 (NPR2) in cumulus cells.
2014,
Pubmed
Zhang,
Brain natriuretic peptide and C-type natriuretic peptide maintain porcine oocyte meiotic arrest.
2015,
Pubmed