Restricted accessResearch articleFirst published online 2018-08
Somatic Angiotensin I-Converting Enzyme Regulates Self-Renewal of Mouse Spermatogonial Stem Cells Through the Mitogen-Activated Protein Kinase Signaling Pathway
Spermatogonial stem cell (SSC) self-renewal is an indispensable part of spermatogenesis. Angiotensin I-converting enzyme (ACE) is a zinc dipeptidyl carboxypeptidase that plays a critical role in the regulation of the renin–angiotensin system. In this study, we used reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis to confirm that somatic ACE (sACE), but not testicular ACE (tACE), is expressed in mouse testis before postpartum day 7 and in cultured SSCs. Our results revealed that sACE is located on the membrane of SSCs. Treating cultured SSCs with the ACE competitive inhibitor captopril was found to inhibit sACE activity, and significantly reduced the proliferation rate of SSCs. Microarray analysis identified 651 genes with significant differential expression. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that these differentially expressed genes are mainly involved in the mitogen-activated protein kinase (MAPK) signaling pathway. sACE was found to play an important role in SSC self-renewal through the regulation of MAPK-dependent cell proliferation.
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References
1.
de RooijDG and RussellLD. (2000). All you wanted to know about spermatogonia but were afraid to ask. J Androl, 21:776–798.
2.
MeachemS, von SchonfeldtV and SchlattS. (2001). Spermatogonia: stem cells with a great perspective. Reproduction, 121:825–834.
3.
Kanatsu-ShinoharaM and ShinoharaT. (2013). Spermatogonial stem cell self-renewal and development. Annu Rev Cell Dev Biol, 29:163–187.
4.
MengX, LindahlM, HyvonenME, ParvinenM, de RooijDG, HessMW, Raatikainen-AhokasA, SainioK, RauvalaH, et al. (2000). Regulation of cell fate decision of undifferentiated spermatogonia by GDNF. Science, 287:1489–1493.
Kanatsu-ShinoharaM, OgonukiN, InoueK, MikiH, OguraA, ToyokuniS and ShinoharaT. (2003). Long-term proliferation in culture and germline transmission of mouse male germline stem cells. Biol Reprod, 69:612–616.
7.
KubotaH, AvarbockMR and BrinsterRL. (2004). Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells. Proc Natl Acad Sci U S A, 101:16489–16494.
8.
OatleyJM, AvarbockMR and BrinsterRL. (2007). Glial cell line-derived neurotrophic factor regulation of genes essential for self-renewal of mouse spermatogonial stem cells is dependent on Src family kinase signaling. J Biol Chem, 282:25842–25851.
9.
OatleyJM, AvarbockMR, TelarantaAI, FearonDT and BrinsterRL. (2006). Identifying genes important for spermatogonial stem cell self-renewal and survival. Proc Natl Acad Sci U S A, 103:9524–9529.
10.
BuaasFW, KirshAL, SharmaM, McLeanDJ, MorrisJL, GriswoldMD, de RooijDG and BraunRE. (2004). Plzf is required in adult male germ cells for stem cell self-renewal. Nat Genet, 36:647–652.
11.
CostoyaJA, HobbsRM, BarnaM, CattorettiG, ManovaK, SukhwaniM, OrwigKE, WolgemuthDJ and PandolfiPP. (2004). Essential role of Plzf in maintenance of spermatogonial stem cells. Nat Genet, 36:653–659.
12.
ZhouQ, GuoY, ZhengB, ShaoB, JiangM, WangG, ZhouT, WangL, ZhouZ, GuoX and HuangX. (2015). Establishment of a proteome profile and identification of molecular markers for mouse spermatogonial stem cells. J Cell Mol Med, 19:521–534.
13.
JokubaitisVJ, SinkaL, DriessenR, WhittyG, HaylockDN, BertoncelloI, SmithI, PeaultB, TavianM and SimmonsPJ. (2008). Angiotensin-converting enzyme (CD143) marks hematopoietic stem cells in human embryonic, fetal, and adult hematopoietic tissues. Blood, 111:4055–4063.
14.
ChisiJE, BriscoeCV, EzanE, GenetR, RichesAC and Wdzieczak-BakalaJ. (2000). Captopril inhibits in vitro and in vivo the proliferation of primitive haematopoietic cells induced into cell cycle by cytotoxic drug administration or irradiation but has no effect on myeloid leukaemia cell proliferation. Br J Haematol, 109:563–570.
15.
MoriS and TokuyamaK. (2007). ACE activity affects myogenic differentiation via mTOR signaling. Biochem Biophys Res Commun, 363:597–602.
16.
KugaevskayaEV, TimoshenkoOS and SolovyevaNI. (2015). Angiotensin converting enzyme: the antigenic properties of the domain, role in Alzheimer's disease and tumor progression. Biomed Khim, 61:301–311.
17.
FrankeFE, PaulsK, MetzgerR and DanilovSM. (2003). Angiotensin I-converting enzyme and potential substrates in human testis and testicular tumours. APMIS, 111:234–243. discussion:243–244.
18.
KondohG, TojoH, NakataniY, KomazawaN, MurataC, YamagataK, MaedaY, KinoshitaT, OkabeM, TaguchiR and TakedaJ. (2005). Angiotensin-converting enzyme is a GPI-anchored protein releasing factor crucial for fertilization. Nat Med, 11:160–166.
19.
HooperNM. (1991). Angiotensin converting enzyme: implications from molecular biology for its physiological functions. Int J Biochem, 23:641–647.
20.
TurnerAJ and HooperNM. (2002). The angiotensin-converting enzyme gene family: genomics and pharmacology. Trends Pharmacol Sci, 23:177–183.
21.
BrownNJ and VaughanDE. (1998). Angiotensin-converting enzyme inhibitors. Circulation, 97:1411–1420.
22.
ZamanMA, OparilS and CalhounDA. (2002). Drugs targeting the renin-angiotensin-aldosterone system. Nat Rev Drug Discov, 1:621–636.
23.
SoubrierF, Alhenc-GelasF, HubertC, AllegriniJ, JohnM, TregearG and CorvolP. (1988). Two putative active centers in human angiotensin I-converting enzyme revealed by molecular cloning. Proc Natl Acad Sci U S A, 85:9386–9390.
24.
SturrockED, NateshR, van RooyenJM and AcharyaKR. (2004). Structure of angiotensin I-converting enzyme. Cell Mol Life Sci, 61:2677–2686.
25.
HagamanJR, MoyerJS, BachmanES, SibonyM, MagyarPL, WelchJE, SmithiesO, KregeJH and O'BrienDA. (1998). Angiotensin-converting enzyme and male fertility. Proc Natl Acad Sci U S A, 95:2552–2557.
26.
FuchsS, FrenzelK, HubertC, LyngR, MullerL, MichaudA, XiaoHD, AdamsJW, CapecchiMR, et al. (2005). Male fertility is dependent on dipeptidase activity of testis ACE. Nat Med, 11:1140–1142. author reply:1142–1143.
27.
HiiSI, NicolDL, GotleyDC, ThompsonLC, GreenMK and JonssonJR. (1998). Captopril inhibits tumour growth in a xenograft model of human renal cell carcinoma. Br J Cancer, 77:880–883.
28.
JohnsonGL and LapadatR. (2002). Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science, 298:1911–1912.
SunY, LiuWZ, LiuT, FengX, YangN and ZhouHF. (2015). Signaling pathway of MAPK/ERK in cell proliferation, differentiation, migration, senescence and apoptosis. J Recept Signal Transduct Res, 35:600–604.
31.
MorimotoH, IwataK, OgonukiN, InoueK, AtsuoO, Kanatsu-ShinoharaM, MorimotoT, Yabe-NishimuraC and ShinoharaT. (2013). ROS are required for mouse spermatogonial stem cell self-renewal. Cell Stem Cell, 12:774–786.
32.
LuZ and XuS. (2006). ERK1/2 MAP kinases in cell survival and apoptosis. IUBMB Life, 58:621–631.
33.
BoucherMJ, MorissetJ, VachonPH, ReedJC, LaineJ and RivardN. (2000). MEK/ERK signaling pathway regulates the expression of Bcl-2, Bcl-X(L), and Mcl-1 and promotes survival of human pancreatic cancer cells. J Cell Biochem, 79:355–369.
34.
JostM, HuggettTM, KariC, BoiseLH and RodeckU. (2001). Epidermal growth factor receptor-dependent control of keratinocyte survival and Bcl-xL expression through a MEK-dependent pathway. J Biol Chem, 276:6320–6326.
35.
HasegawaK, NamekawaSH and SagaY. (2013). MEK/ERK signaling directly and indirectly contributes to the cyclical self-renewal of spermatogonial stem cells. Stem Cells, 31:2517–2527.
36.
BeenkenA and MohammadiM. (2009). The FGF family: biology, pathophysiology and therapy. Nat Rev Drug Discov, 8:235–253.
37.
RusnatiM and PrestaM. (2007). Fibroblast growth factors/fibroblast growth factor receptors as targets for the development of anti-angiogenesis strategies. Curr Pharm Des, 13:2025–2044.
38.
Keren-PolitanskyA, KerenA and BengalE. (2009). Neural ectoderm-secreted FGF initiates the expression of Nkx2.5 in cardiac progenitors via a p38 MAPK/CREB pathway. Dev Biol, 335:374–384.
39.
WeisingerK, KayamG, Missulawin-DrillmanT and Sela-DonenfeldD. (2010). Analysis of expression and function of FGF-MAPK signaling components in the hindbrain reveals a central role for FGF3 in the regulation of Krox20, mediated by Pea3. Dev Biol, 344:881–895.
40.
HasegawaK and SagaY. (2014). FGF8-FGFR1 signaling acts as a niche factor for maintaining undifferentiated spermatogonia in the mouse. Biol Reprod, 91:145.
41.
ShinyaM, KoshidaS, SawadaA, KuroiwaA and TakedaH. (2001). Fgf sinalling through MAPK cascade is required for development of the subpallial telencephalon in zebrafish embryos. Development, 128:4153–4164.
42.
WalsheJ, MaroonH, McGonnellIM, DicksonC and MasonI. (2002). Establishment of hindbrain segmental identity requires signaling by FGF3 and FGF8. Curr Biol, 12:1117–1123.
43.
MavesL, JackmanW and KimmelCB. (2002). FGF3 and FGf8 mediate a rhombomere 4 signaling activity in the zebrafish hindbrain. Development, 129:3825–3837.
44.
CauntCJ, RiversCA, Conway-CampbellBL, NormanMR and McArdleCA. (2008). Epidermal growth factor receptor and protein kinase C signaling to ERK2: spatiotemporal regulation of ERK2 by dual specificity phosphatases. J Biol Chem, 283:6241–6252.
45.
PattersonKI, BrummerT, O'BrienPM and DalyRJ. (2009). Dual-specificity phosphatases: critical regulators with diverse cellular targets. Biochem J, 418:475–489.
46.
GrumontRJ, RaskoJE, StrasserA and GerondakisS. (1996). Activation of the mitogen-activated protein kinase pathway induces transcription of the PAC-1 phosphatase gene. Mol Cell Biol, 16:2913–2921.
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