Role of Dicer1-Dependent Factors in the Paracrine Regulation of Epididymal Gene Expression.
Dicer1 is an endoribonuclease involved in the biogenesis of functional molecules such as microRNAs (miRNAs) and endogenous small interfering RNAs (endo-siRNAs). These small non-coding RNAs are important regulators of post-transcriptional gene expression and participate in the control of male fertility. With the knowledge that 1) Dicer1-dependent factors are required for proper sperm maturation in the epididymis, and that 2) miRNAs are potent mediators of intercellular communication in most biological systems, we investigated the role of Dicer1-dependent factors produced by the proximal epididymis (initial segment/caput)- including miRNAs- on the regulation of epididymal gene expression in the distal epididymis regions (i.e. corpus and cauda). To this end, we performed comparative microarray and ANOVA analyses on control vs. Defb41iCre/wt;Dicer1fl/fl mice in which functional Dicer1 is absent from the principal cells of the proximal epididymis. We identified 35 and 33 transcripts that displayed significant expression level changes in the corpus and cauda regions (Fold change > 2 or < -2; p < 0.002), respectively. Among these transcripts, Zn-alpha 2-glycoprotein (Azgp1) encodes for a sperm equatorial protein whose expression in the epididymis of Dicer1 cKO mice is significantly increased compared to controls. In addition, 154 miRNAs, including miR-210, miR-672, miR-191 and miR-204, showed significantly impaired biogenesis in the absence of Dicer1 from the principal cells of the proximal epididymis (Fold change > 2 or < -2; p < 0.01). These miRNAs are secreted via extracellular vesicles (EVs) derived from the DC2 epididymal principal cell line, and their expression correlates with target transcripts involved in distinct biological pathways, as evidenced by in silico analysis. Albeit correlative and based on in silico approach, our study proposes that Dicer1-dependent factors trigger- directly or not-significant genes expression changes in distinct regions of this organ. The paracrine control of functions important to post-testicular sperm maturation by Dicer1-dependent factors may open new avenues for the identification of molecular targets important to male fertility control.
Department of Obstetrics, Gynecology and Reproduction, Université Laval, CHU de Québec Research Center (CHUL), Quebec City, Quebec, Canada.
Olivia
O
Jerczynski
Department of Obstetrics, Gynecology and Reproduction, Université Laval, CHU de Québec Research Center (CHUL), Quebec City, Quebec, Canada.
Nicolas
N
Lacroix-Pépin
Department of Immunity and Infectious Diseases, Université Laval, CHU de Québec Research Center (CHUL), Quebec City, Quebec, Canada.
Eric
E
Boilard
Endocrinology unit, CHU de Québec Research Center (CHUL), Quebec City, Quebec, Canada.
Ezequiel
E
Calvo
Department of Obstetrics, Gynecology and Reproduction, Université Laval, CHU de Québec Research Center (CHUL), Quebec City, Quebec, Canada.
Agathe
A
Bernet
Department of Obstetrics, Gynecology and Reproduction, Université Laval, CHU de Québec Research Center (CHUL), Quebec City, Quebec, Canada.
Michel A
MA
Fortier
Department of Physiology, Institute of Biomedicine, University of Turku, Turku, Finland.
Ida
I
Björkgren
Department of Physiology, Institute of Biomedicine, University of Turku, Turku, Finland.|Turku Center for Disease Modeling, University of Turku, Turku, Finland.
Petra
P
Sipilä
Department of Obstetrics, Gynecology and Reproduction, Université Laval, CHU de Québec Research Center (CHUL), Quebec City, Quebec, Canada.
Clémence
C
Belleannée
D000818:Animals; D053487:DEAD-box RNA Helicases / Q000096:biosynthesis* / Q000235:genetics; D004822:Epididymis / Q000254:growth & development* / Q000378:metabolism; D005298:Fertility / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D043244:Ribonuclease III / Q000096:biosynthesis* / Q000235:genetics; D013079:Sperm Maturation / Q000235:genetics*; D013094:Spermatozoa / Q000254:growth & development / Q000378:metabolism
D006023:Glycoproteins; D035683:MicroRNAs; C509781:Dicer1 protein, mouse; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases
Int J Androl. 2011 Oct;34(5 Pt 2):e475-86
21875428
Nat Cell Biol. 2011 Apr;13(4):423-33
21423178
Science. 2016 Jan 22;351(6271):391-396
26721685
J Extracell Vesicles. 2015 Nov 11;4:28680
26563734
J Extracell Vesicles. 2013 Sep 12;2:
24223256
Hum Reprod. 2013 Jun;28(6):1455-67
23539611
Reprod Biol Endocrinol. 2011 Sep 26;9(1):128
21942998
J Physiol. 2012 Sep 1;590(17):4209-22
22711960
Mol Reprod Dev. 2007 Sep;74(9):1124-31
17393427
Biol Reprod. 2015 Oct;93(4):91
26333995
Cell Res. 2009 Mar;19(3):282-95
19188930
Am J Physiol Cell Physiol. 2010 Apr;298(4):C817-30
20071692
F1000 Biol Rep. 2011;3:15
21876726
Clin Chem. 2010 Nov;56(11):1733-41
20847327
PLoS Genet. 2014 Oct 16;10(10):e1004597
25329700
PLoS One. 2014 Sep 22;9(9):e107023
25244517
Cell. 2013 Aug 29;154(5):1010-1022
23993094
PLoS One. 2011;6(9):e24821
21949761
Dev Biol. 2012 Sep 1;369(1):43-53
22750823
Spermatogenesis. 2012 Jan 1;2(1):1-5
22553484
Methods Mol Biol. 2010;631:109-22
20204872
J Intern Med. 2012 Apr;271(4):400-13
22112042
Proc Natl Acad Sci U S A. 2011 Mar 22;108(12):5003-8
21383194
Cell Cycle. 2012 Jul 1;11(13):2495-506
22739938
Nat Rev Mol Cell Biol. 2012 Apr 18;13(5):328-35
22510790
Nature. 2010 Jun 3;465(7298):584-9
20424607
FASEB J. 2015 Feb;29(2):433-42
25366345
Curr Opin Cell Biol. 2014 Aug;29:116-25
24959705
Org Biomol Chem. 2015 Oct 14;13(38):9775-82
26264754
J Androl. 2002 Nov-Dec;23(6):854-69
12399533
Angew Chem Int Ed Engl. 2007;46(37):6985-94
17712806
J Androl. 2012 Sep-Oct;33(5):963-74
22207704
Biol Reprod. 2006 Jun;74(6):1104-13
16510839
J Cell Biochem. 2009 Apr 15;106(6):957-66
19204935
Cell Death Differ. 2004 Aug;11(8):943-5
15031725
Asian J Androl. 2015 Sep-Oct;17(5):726-9
26112475
Prostate. 2012 Dec 1;72(16):1736-45
22539202
Nat Cell Biol. 2007 Jun;9(6):654-9
17486113
J Cell Physiol. 2012 Dec;227(12):3876-86
22552861
Biol Reprod. 2012 Oct 18;87(4):94
22875906
PLoS One. 2015 Jan 14;10(1):e0116812
25587983
PLoS One. 2011;6(10):e25241
21998645
Mol Reprod Dev. 2010 Jun;77(6):550-63
20422713
Nucleic Acids Res. 2001 May 1;29(9):e45
11328886
Cell. 2009 Jan 23;136(2):215-33
19167326
Hum Reprod Update. 2009 Mar-Apr;15(2):213-27
19136456
Curr Opin Cell Biol. 2004 Oct;16(5):477-85
15363796
Arterioscler Thromb Vasc Biol. 2013 Feb;33(2):186-92
23325475
Biol Reprod. 1969 Jun;1:Suppl 1:155-75
5406325
J Acquir Immune Defic Syndr. 2015 Nov 1;70(3):219-27
26181817
J Extracell Vesicles. 2015 May 14;4:27066
25979354
Mol Reprod Dev. 2002 Nov;63(3):394-410
12237956
J Reprod Immunol. 2015 Aug;110:117-29
25899154
Nat Commun. 2014 Feb 28;5:3292
24576947
J Cell Sci. 2013 Dec 1;126(Pt 23):5513-23
24105266
Biol Reprod. 2008 Oct;79(4):696-703
18633141
PLoS One. 2015 Aug 13;10(8):e0135605
26270822
Andrologia. 1996;28 Suppl 1:57-9
9017097
J Extracell Vesicles. 2014 Sep 30;3:
25317276
PLoS One. 2016 Oct 3;11(10):e0163876
27695046
Reproduction. 2013 Jun 14;146(1):R21-35
23613619
PLoS One. 2012;7(6):e38457
22701646
Development. 2016 Feb 15;143(4):635-47
26718009
PLoS One. 2013 Jun 13;8(6):e65364
23785420
Biol Reprod. 2013 Aug 15;89(2):30
23803555
Cell Tissue Res. 2012 Sep;349(3):717-31
22427067
EMBO Mol Med. 2013 Feb;5(2):235-49
23165896
Nat Cell Biol. 2009 Mar;11(3):228-34
19255566
PLoS One. 2008 Mar 05;3(3):e1738
18320056
Proc Natl Acad Sci U S A. 2014 Jul 15;111(28):E2851-7
24982181
Cell. 2014 Nov 20;159(5):1153-1167
25416952