Coding and Non-Coding RNAs, as Male Fertility and Infertility Biomarkers

Document Type : Review Article


1 Men’s Health and Reproductive Health Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

2 Department of Cellular and Molecular Sciences, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

3 Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

4 Department of Biology, Faculty of Biological Sciences, Sabzevar Branch, Islamic Azad University, Sabzevar, Iran

5 Non-Communicable Disease Control Department, Public Health Department, Minis try of Health and Medical Education, Tehran, Iran


Semen analysis is usually the first step in the assessment of male fertility. Although analyzes provide valuable information about male fertility, success of cytoplasmic sperm injection using this method is not predictable. In the recent years, studies have shown that sperm quality assessment helps clinicians predict male fertility status based on the expression of biomarkers. To write this article, a comprehensive study was conducted on several RNA transcripts by searching related words on medical information databases by 2018. According to the literature, spermatogenesis based disorders in male infertility have a significant relationship with the expression level of some RNA molecules (like DAZ and PRM1/PRM2 ratio) in semen and testicular tissue. Thus, they might be used as predictor biomarkersto evaluate success rate of testicular sperm extraction (TESE) procedure, but confirmation of this hypothesis requires more extensive research. By comparing the number of RNAs attributed to each fertility disorder in men, it is possible to trace the causes of disease or return fertility to some infertile patients by regulating the mentioned molecules. Further researches can provide a better understanding of the use of RNA expression profiles in the diagnosis and treatment of male infertility.


1. Chen Q, Yan W, Duan E. Epigenetic inheritance of acquired traits through sperm RNAs and sperm RNA modifications. Nat Rev Gen­et. 2016; 17(12): 733-743.
2. Li C, Zhou X. Gene transcripts in spermatozoa: Markers of male infertility. Clin Chim Acta. 2012; 413(13-14): 1035-1038.
3. Krausz C, Escamilla AR, Chianese C. Genetics of male infertility: from research to clinic. Reproduction. 2015; 150(5): R159-R174.
4. Montjean D, De La Grange P, Gentien D, Rapinat A, Belloc S, Cohen-Bacrie P, et al. Sperm transcriptome profiling in oligozoo­spermia. J Assis t Reprod Genet. 2012; 29(1): 3-10.
5. Aslani F, Modarresi MH, Soltanghoraee H, Akhondi MM, Shabani A, Lakpour N, et al. Seminal molecular markers as a non-invasive diagnos tic tool for the evaluation of spermatogenesis in non-ob­s tructive azoospermia. Sys t Biol Reprod Med. 2011; 57(4): 190- 196.
6. Cas tillo J, Jodar M, Oliva R. The contribution of human sperm pro­teins to the development and epigenome of the preimplantation embryo. Hum Reprod Update. 2018; 24(5): 535-555.
7. Rahiminia T, Yazd EF, Fesahat F, Moein MR, Mirjalili AM, Talebi AR. Sperm chromatin and DNA integrity, methyltransferase mRNA lev­els, and global DNA methylation in oligoas thenoteratozoospermia. Clin Exp Reprod Med. 2018; 45(1): 17-24.
8. Heidari S, Taromchi A, Nejatbakhsh R, Shokri S. Expression and localisation of RXFP 3 in human spermatozoa and impact of INSL 7 on sperm functions. Andrologia. 2018; 50(3): e12928.
9. Aghajanpour S, Ghaedi K, Salamian A, Deemeh M, Tavalaee M, Moshtaghian J, et al. Quantitative expression of phospholipase C zeta, as an index to assess fertilization potential of a semen sam­ple. Hum Reprod. 2011; 26(11): 2950-2956.
10. Yes te M, Jones C, Amdani SN, Yelumalai S, Mounce G, da Silva SJM, et al. Does advancing male age influence the expression lev­els and localisation patterns of phospholipase C zeta (PLCζ) in hu­man sperm? Sci Rep. 2016; 6: 27543.
11. Javadian-Elyaderani S, Ghaedi K, Tavalaee M, Rabiee F, Deemeh MR, Nasr-Esfahani MH. Diagnosis of genetic defects through par­allel assessment of PLCζ and CAPZA3 in infertile men with his tory of failed oocyte activation. Iran J Basic Med Sci. 2016; 19(3): 281-289.
12. Abadi MKD, Tavalaee M, Shahverdi A, Nasr-Esfahani MH. Evalua­tion of PLCζ and PAWP expression in globozoospermic individuals. Cell J. 2016; 18(3): 438-445.
13. Tavalaee M, Nasr-Esfahani MH. Expression profile of PLC ζ, PAWP, and TR-KIT in association with fertilization potential, embryo devel­opment, and pregnancy outcomes in globozoospermic candidates for intra-cytoplasmic sperm injection and artificial oocyte activation. Andrology. 2016; 4(5): 850-856.
14. Freour T, Barragan M, Ferrer-Vaquer A, Rodríguez A, Vassena R. WBP2NL/PAWP mRNA and protein expression in sperm cells are not related to semen parameters, fertilization rate, or reproductive outcome. J Assis Reprod Genet. 2017; 34(6): 803-810.
15. Muciaccia B, Sette C, Paronetto MP, Barchi M, Pensini S, D'agos tino A, et al. Expression of a truncated form of KIT tyrosine kinase in human spermatozoa correlates with sperm DNA integrity. Hum Reprod. 2010; 25(9): 2188-2202.
16. Eelaminejad Z, Favaedi R, Sodeifi N, Sadighi Gilani MA, Shah­hoseini M. Deficient expression of JMJD1A his tone demethylase in patients with round spermatid maturation arres t. Reprod Biomed Online. 2017; 34(1): 82-89.
17. Moghbelinejad S, Najafipour R, Samimi Hashjin A. Comparison of protamine 1 to protamine 2 mRNA ratio and YBX2 gene mRNA content in tes ticular tissue of fertile and azoospermic men. Int J Fertil S teril. 2015; 9(3): 338-345.
18. Najafipour R, Moghbelinejad S, Samimi Hashjin A, Rajaei F, Rash­vand Z. Evaluation of mRNA contents of YBX2 and JHDM2A genes on tes ticular tissues of azoospermic men with different classes of spermatogenesis. Cell J. 2015; 17(1): 121-128.
19. Flannigan R, Mielnik A, Robinson BD, Khani F, Bolyakov A, Schlegel PN, et al. YBX2 dysregulation in maturation arres t NOA YBX2 dysregulation is a potential cause for late maturation arres t in men with non-obs tructive azoospermia. bioRxiv. 2018: Available from: (12 Mar 2020).
20. Savadi-Shiraz E, Edalatkhah H, Talebi S, Heidari-Vala H, Zande­mami M, Pahlavan S, et al. Quantification of sperm specific mRNA transcripts (PRM1, PRM2, and TNP2) in teratozoospermia and normozoospermia: New correlations between mRNA content and morphology of sperm. Mol Reprod Dev. 2015; 82(1): 26-35.
21. S teger K, Fink L, Failing K, Bohle RM, Kliesch S, Weidner W, et al. Decreased protamine-1 transcript levels in tes tes from infertile men. Mol Hum Reprod. 2003; 9(6): 331-336.
22. Aboutorabi R, Asghari M, Bakhteyari A, Baghazadeh S, Mos tafavi FS. PROTAMINE1 and PROTAMINE2 genes expression in the sperms of oligoas thenospermic individuals and intrauterine insemi­nation candidates couples: is there any significant differences? Adv Biomed Res. 2016; 5; 164.
23. Zalata AA, Mokhtar N, Atwa A, Khaled M, Shaker OG. The role of protamine 2 gene expression and caspase 9 activity in male infertil­ity. J Urol. 2016; 195(3): 796-800.
24. Javadirad SM, Hojati Z, Ghaedi K, Nasr-Esfahani MH. Expression ratio of his tone demethylase KDM 3A to protamine-1 mRNA is pre­dictive of successful tes ticular sperm extraction in men with ob­s tructive and non-obs tructive azoospermia. Andrology. 2016; 4(3): 492-499.
25. Depa-Martynów M, Kempis ty B, Lianeri M, Jagodziński PP, Jedrze­jczak P. Association between fertilin beta, protamines 1 and 2 and spermatid-specific linker his tone H1-like protein mRNA levels, fer­tilization ability of human spermatozoa, and quality of preimplanta­tion embryos. Folia His tochem Cyto. 2007; 45 Suppl 1: S79-85.
26. Hashemi MS, Mozdarani H, Ghaedi K, Nasr-Esfahani MH. Among seven tes tis-specific molecular markers, SPEM 1 appears to have a significant clinical value for prediction of sperm retrieval in azoo­spermic men. Andrology. 2018; 6(6): 890-895.
27. Kumar K, Deka D, Singh A, Chattopadhyay P, Dada R. Expression pattern of PRM2, HSP90 and WNT5A in male partners of couples experiencing idiopathic recurrent miscarriages. J Genet. 2012; 91(3): 363-366.
28. Becker S, Soffer Y, Lewin LM, Yogev L, Shochat L, Golan R. Sper­miogenesis defects in human: detection of transition proteins in se­men from some infertile men. Andrologia. 2008; 40(4): 203-208.
29. Motiei M, Tavalaee M, Rabiei F, Hajihosseini R, Nasr-Esfahani MH. Evaluation of HSPA 2 in fertile and infertile individuals. Andrologia. 2013; 45(1): 66-72.
30. Aarabi M, Ousati-Ashtiani Z, Nazarian A, Modarressi MH, Heidari M. Association of TGIFLX/Y mRNA expression with azoospermia in infertile men. Mol Reprod Dev. 2008; 75(12): 1761-1766.
31. Aarabi M, Modarressi MH, Soltanghoraee H, Behjati R, Amirjannati N, Akhondi MM. Tes ticular expression of synaptonemal complex protein 3 (SYCP3) messenger ribonucleic acid in 110 patients with nonobs tructive azoospermia. Fertil S teril. 2006; 86(2): 325-331.
32. Shafipour M, Sabbaghian M, Shahhoseini M, Sadighi Gilani MA. Comparative expression analysis of Septin 14 in tes tes of infertile men with normal spermatogenesis and spermatogenic failure. Iran J Reprod Med. 2014; 12(3): 205-208.
33. Aarabi M, Saliminejad K, Sadeghi M, Soltanghoraee H, Amirjannati N, Modarressi M. Deletion and tes ticular expression of DAZ (de­leted in azoospermia) gene in patients with non-obs tructive azoo­spermia. Iran J Public Health. 2009; 38(3): 17-23.
34. Aarabi M, Soltanghoraee H, Amirjannati N, Ghaffari M, Sadeghi MR, Akhondi MM, et al. Tes tis specific gene 10 expression in the tes tes of patients with non-obs tructive azoospermia. J Reprod In­fertil. 2006; 7(3): 179-186.
35. Mamaghani AM, Hosseini SJ, Moslemi E. Hormonal profiling and clus terin gene expression in non-obs tructive azoospermic patients. Tehran Univ Med J. 2018;75(11): 819-827.
36. Faghihi Zohani F, Shahhoseini M, Favaedi R, Sadighi Gilani MA, Shahzadeh Fazeli A. Gene expression analysis of H2B his tone variant hTSH2B in tes tis tissues of non-obs tructive azoospermic patients referred to Royan Ins titute. Iran J Reprod Med. 2013; 44-45.
37. Taheri H, Salehi M, Eslami Arshaghi T, Mosahebi M. S tudying of the expression of BAX and BCL-2 Genes in men with sperm DNA fragmentation based on DFI and comet assay. ASJ. 2015; 12(1): 29-36.
38. Guido C, Perrotta I, Panza S, Middea E, Avena P, Santoro M, et al. Human sperm physiology: Es trogen receptor alpha (ERα) and es­trogen receptor beta (ERβ) influence sperm metabolism and may be involved in the pathophysiology of varicocele-associated male infertility. J Cell Physiol. 2011; 226(12): 3403-3412.
39. Abhari A, Zarghami N, Farzadi L, Nouri M, Shahnazi V. Altered of microRNA expression level in oligospermic patients. Iran J Reprod Med. 2014; 12(10): 681-686.
40. Jodar M, Selvaraju S, Sendler E, Diamond MP, Krawetz SA, Net­work RM. The presence, role and clinical use of spermatozoal RNAs. Hum Reprod Update. 2013; 19(6): 604-624.
41. Azad N, Nazarian H, Nazari L, Ghaffari Novin M, Piryaei A, Heidari MH, et al. Evaluation of PAWP and PLC? Expression in infertile men with previous ICSI fertilization failure. Urol J. 2018; 15(3): 116-121.
42. Park JH, Kim SK, Kim J, Kim JH, Chang JH, Jee BC, et al. Rela­tionship between phospholipase C zeta immunoreactivity and DNA fragmentation and oxidation in human sperm. Obs tet Gynecol Sci. 2015; 58(3): 232-238.
43. Heytens E, Parrington J, Coward K, Young C, Lambrecht S, Yoon SY, et al. Reduced amounts and abnormal forms of phospholipase C zeta (PLCζ) in spermatozoa from infertile men. Hum Reprod. 2009; 24(10): 2417-2428.
44. Ghazavi-Khorasgani N, Janghorban-Laricheh E, Tavalaee M, Zohrabi D, Abbasi H, Nasr-Esfahani MH. Evaluation of pos t-acro­somal sheath WW domain binding protein expression in sperma­tozoa from infertile men with varicocele. Tehran Univ Med J. 2017; 75(6): 417-423.
45. Almabhouh FA, Singh HJ. Adverse effects of leptin on his tone-to-protamine transition during spermatogenesis are prevented by me­latonin in Sprague-Dawley rats. Andrologia. 2018; 50(1).
46. Lambard S, Galeraud-Denis I, Martin G, Levy R, Chocat A, Car­reau S. Analysis and significance of mRNA in human ejaculated sperm from normozoospermic donors: relationship to sperm motil­ity and capacitation. Mol Hum Reprod. 2004; 10(7): 535-541.
47. Rogenhofer N, Ott J, Pilatz A, Wolf J, Thaler CJ, Windischbauer L, et al. Unexplained recurrent miscarriages are associated with an aberrant sperm protamine mRNA content. Hum Reprod. 2017; 32(8): 1574-1582.
48. Liu T, Huang Y, Liu J, Zhao Y, Jiang L, Huang Q, et al. MicroR­NA-122 influences the development of sperm abnormalities from human induced pluripotent s tem cells by regulating TNP2 expres­sion. S tem Cell Dev. 2013; 22(12): 1839-1850.
49. Youssefian L, Vahidnezhad H, Raoofian R, Greene WK, Heidari M. Potential functions of the human homeobox TGIFLX/Y genes in normal and abnormal development. Eur J Clin Med Oncol. 2012; 4(1): 23-26.
50. Aydos OSE, Hekmatshoar Y, Altınok B, Özkan T, Şakirağaoğlu O, Karadağ A, et al. Genetic polymorphisms in PRM1, PRM2, and YBX2 genes are associated with male factor infertility. Genetic Tes t Mol Biomarkers. 2018; 22(1): 55-61.
51. Moghbelinejad S, Mozdarania H, Ghoraeian P, Asadi R. Basic and clinical genetic s tudies on male infertility in Iran during 2000-2016: a review. Int J Reprod Biomed. 2018; 16(3): 131-148.
52. Hammoud S, Emery BR, Dunn D, Weiss RB, Carrell DT. Sequence alterations in the YBX2 gene are associated with male factor infer­tility. Fertil S teril. 2009; 91(4): 1090-1095.
53. Azizi F, Ghafouri-Fard S. Outer dense fiber proteins: bridging between male infertility and cancer. Arch Iran Med. 2017; 20(5): 320-325.
54. Etheridge A, Lee I, Hood L, Galas D, Wang K. Extracellular microR­NA: a new source of biomarkers. Mutat Res. 2011; 717(1-2): 85-90.
55. Azizi F, Omrani MD, Sadighi Gilani MA, Hosseini J. The Genetic causes of male infertility in Iranian population; a sys tematic review. Men's Health Journal. 2018; 2(1): e1.
56. Abhari A, Zarghami N, Shahnazi V, Barzegar A, Farzadi L, Karami H, et al. Significance of microRNA targeted es trogen receptor in male fertility. Iran J Basic Med Sci. 2014; 17(2): 81-86.
57. Zhao Y, Deng C, Wang J, Xiao J, Gatalica Z, Recker RR, et al. Let-7 family miRNAs regulate es trogen receptor alpha signaling in es trogen receptor positive breas t cancer. Breas t Cancer Res Treat. 2011; 127(1): 69-80.
58. Rahbar S, Novin MG, Alizadeh E, Shahnazi V, Pashaei-Asl F, Asr­Badr YA, et al. New insights into the expression profile of MicroR­NA-34c and P53 in infertile men spermatozoa and tes ticular tissue. Cell Mol Biol (Noisy-le-grand). 2017; 63(8): 77-83.
59. Bouhallier F, Allioli N, Lavial F, Chalmel F, Perrard MH, Durand P, et al. Role of miR-34c microRNA in the late s teps of spermatogen­esis. RNA. 2010; 16(4): 720-731.
60. Mukherjee A, Koli S, Reddy KVR. Regulatory non-coding tran­scripts in spermatogenesis: shedding light on 'dark matter'. Androl­ogy. 2014; 2(3): 360-369.
61. Kamaliyan Z, Pouriamanesh S, Soosanabadi M, Gholami M, Mir­fakhraie R. Inves tigation of piwi-interacting RNA pathway genes role in idiopathic non-obs tructive azoospermia. Sci Rep. 2018; 8(1): 142.
62. Gu A, Ji G, Shi X, Long Y, Xia Y, Song L, et al. Genetic variants in Piwi-interacting RNA pathway genes confer susceptibility to sper­matogenic failure in a Chinese population. Hum Reprod. 2010; 25(12): 2955-2961.
63. Vourekas A, Zheng K, Fu Q, Maragkakis M, Alexiou P, Ma J, et al. The RNA helicase MOV10L1 binds piRNA precursors to initiate piRNA processing. Genes Dev. 2015; 29(6): 617-629.
64. Sarkardeh H, Totonchi M, Asadpour O, Sadighi Gilani MA, Zamani Es teki M, Almadani N, et al. Association of MOV10L1 gene poly­morphisms and male infertility in azoospermic men with complete maturation arres t. J Assis t Reprod Genet. 2014; 31(7): 865-871.
65. Zhang L, Liu Z, Li X, Zhang P, Wang J, Zhu D, et al. Low long non-coding RNA HOTAIR expression is associated with down-regulation of Nrf2 in the spermatozoa of patients with as thenozoo­spermia or oligoas thenozoospermia. Int J Clin Exp Pathol. 2015; 8(11): 14198-14205.
66. Rinn JL, Kertesz M, Wang JK, Squazzo SL, Xu X, Brugmann SA, et al. Functional demarcation of active and silent chromatin domains in human HOX loci by noncoding RNAs. Cell. 2007; 129(7): 1311-1323.
67. Burl RB, Clough S, Sendler E, Es till M, Krawetz SA. Sperm RNA elements as markers of health. Sys t Biol Reprod Med. 2018; 64(1): 25-38.
68. Cao C, Wen Y, Wang X, Fang N, Yuan S, Huang X. Tes ticular piRNA profile comparison between successful and unsuccessful micro- TESE retrieval in NOA patients. J Assis t Reprod Genet. 2018; 35(5): 801-808.