Please wait a minute...
Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2015, Vol. 9 Issue (3) : 361-367     DOI: 10.1007/s11684-015-0395-x
RESEARCH ARTICLE |
Immunohistochemical measurement and expression of Mcl-1 in infertile testes
Raafat Hegazy1,Abdelmonem Hegazy2,*(),Mustafa Ammar3,Emad Salem3
1. Pathology
2. Anatomy and Embryology
3. Urology of Departments, Faculty of Medicine, Zagazig University, Zagazig 44159, Egypt
Download: PDF(1962 KB)   HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks
Abstract  

The diagnosis of azoospermia represents a major challenge to andrologists as this condition may occur despite normal spermatogenesis and genital tracts. Myeloid cell leukemia-1 (Mcl-1) is a member of the Bcl-2 family of proteins involved in regulation of apoptosis in various cell types. This study aimed to investigate the immunohistochemical expression of Mcl-1 in testicular biopsies of subjects with azoospermia. Eighty-six cases with azoospermia were obtained from 509 infertile patients admitted to the Andrology Unit of the Zagazig University Hospitals from January 2010 to December 2011. Biopsies were diagnosed and classified using H&E-stained slide sections. The specimens were subjected to immunohistochemical staining for Mcl-1 and examined through light microscopy. Forty-five cases of maturation arrest (25 at spermatids and 20 at the spermatocytes), 31 cases of hypospermatogenesis (20 moderate and 11 severe), 5 cases of Sertoli?cell-only syndrome, 2 cases of basement membrane hyalinization, and 1 case of tubular and peritubular sclerosis were observed. Normal spermatogenesis was detected in 2 cases. A strong positive immunoreaction in Leydig cells was observed among all investigated specimens. A moderate reaction was detected in spermatocytes and spermatozoa in cases of normal spermatogenesis and hypospermatogenesis, but a negative reaction was detected in cases of maturation arrest and germ cell aplasia. Apoptosis was found to be associated with decreased rate of spermatogenesis. High apoptosis rates may result in azoospermia, which can occur despite normal spermatogenesis and absence of duct obstruction.

Keywords spermatogenesis      testis      Mcl-1 expression      immunohistochemical study      infertility     
Corresponding Authors: Abdelmonem Hegazy   
Just Accepted Date: 29 May 2015   Online First Date: 18 June 2015    Issue Date: 26 August 2015
URL:  
http://academic.hep.com.cn/fmd/EN/10.1007/s11684-015-0395-x     OR     http://academic.hep.com.cn/fmd/EN/Y2015/V9/I3/361
Item Comment
Age 22?35?years
Infertility type Primary infertility
Hormonal profile Normal level
Semen analysis Azoospermia
Size of the testis Normal
Tab.1  Clinicopathological data of the patients
Diagnosis No. %
Normal spermatogenesis 2 2.32%
Moderate hypospermatogenesis 20 23.25%
Severe hypospermatogenesis 11 12.79%
Maturation arrest at the level of spermatids 25 29.06%
Maturation arrest at the level of secondary or primary spermatocytes 20 23.25%
Sertoli cell-only syndrome or partial Sertoli cell-only syndrome 5 5.81%
Basement membrane hyalinization and peritubular fibrosis 2 2.32%
Tubular and peritubular sclerosis 1 1.16%
Total 86 100%
Tab.2  Histopathological examination of the testicular biopsy
Histopathological results Percentage of the immunoreactive (positive) cells
Diagnosis No. of cases Leydig cells Sertoli cells Primary spermatocytes Secondary spermatocytes Spermatids Spermatozoa
Normal spermatogenesis 2 35% 10% 50% 45% 25% 25%
Moderate hypospermatogenesis 15 30% 10% 40% 45% 10% 10%
5 30% 10% 10% 10% 10% 10%
Severe hypospermatogenesis 11 45% 10% 15% 20% 8% 7%
Maturation arrest at the level of spermatids 25 50% 15% 10% 8% 6% No
Maturation arrest at the level of secondary or primary spermatocytes 20 80% 15% 10% 7% No No
Sertoli cell-only syndrome Partial Sertoli cell-only syndrome 3 80% 10% No No No No
2 80% 40% No No No No
Basement membrane hyalinization and peritubular fibrosis 2 10% 10% No No No No
Tubular and peritubular sclerosis 1 No No No No No No
Total 86
Tab.3  Immunohistochemical expression of Mcl-1 in testicular biopsy specimens in relation to histopathological diagnosis
Fig.1  A case of normal spermatogenesis. Numerous sperms (blue arrow), spermatids (yellow arrow), secondary spermatocytes (red arrow), primary spermatocytes (white arrow), Sertoli cells (black arrow), and Leydig cells (green arrow) are shown. H&E stain, 400×.
Fig.2  A case of mild hypospermatogenesis with few sperms (red arrows) and spermatids (blue arrows). H&E stain, 400×.
Fig.3  A case of moderate hypospermatogenesis with strong immunoreactivity to Mcl-1 in spermatids (red arrow) and negative reactions in the hardly visible sperm (blue arrow). 400×.
Fig.4  A case of moderate hypospermatogenesis with moderate positive reactions to Mcl-1 of membranous staining in secondary spermatocytes (yellow arrows) and nuclear staining in spermatids (red arrows) and negative reactions in the spermatozoa (blue arrows). 400×.
Fig.5  A case of severe hypospermatogenesis. Only one sperm cell is visible (blue arrow). H&E stain, 400×.
Fig.6  A case of severe hypospermatogenesis with moderate reactivity for Mcl-1 in spermatocytes (blue arrows) and negative reactivity in spermatids (black arrows) and sperm (red arrow). 400×.
Fig.7  A case of severe hypospermatogenesis with moderate reactivity to Mcl-1 in Leydig cells (red arrows) and negative reactivity in spermatocytes (blue arrows). 400×.
Fig.8  A case of maturation arrest at the spermatids (blue arrows) level. H&E stain, 400×.
Fig.9  A case of spermatogenic arrest with strong cytoplasmic and/or membranous positive reactions in Leydig cells (red arrows) and weak membranous positive reactions in primary spermatocytes (blue arrows). 400×.
Fig.10  A case of Sertoli cell-only syndrome. The tubules are lined by Sertoli cells (red arrow) alone. Leydig cell hyperplasia (blue arrows) is noted. H&E stain, 400×.
Fig.11  A case of Sertoli cell-only syndrome with strong cytoplasmic and/or membranous positive reactions to Mcl-1 in Leydig cells (red arrows) and negative reactions in Sertoli cells (blue arrows). 400×.
Fig.12  A case of Sertoli cell-only syndrome with weak immunoreactivity to Mcl-1 in Sertoli cells (blue arrows). 400×.
Fig.13  A case of Sertoli cell-only syndrome. The tubules are lined by Sertoli cells (blue arrows) alone with moderate peritubular fibrosis, thickening of basement membrane (yellow arrows), and interstitial Leydig cell hyperplasia (white arrow). H&E stain, 400×.
Fig.14  A case of Sertoli cell-only syndrome with strong cytoplasmic and/or membranous positive reactions to Mcl-1 in Sertoli cells (red arrows). Thick peritubular fibrosis (blue arrows) is shown. 400×.
1 Hetts SW. To die or not to die: an overview of apoptosis and its role in disease. JAMA 1998; 279(4): 300–307
doi: 10.1001/jama.279.4.300 pmid: 9450715
2 Kerr JFR, Winterford CM, Harmon BV. Apoptosis. Its significance in cancer and cancer therapy. Cancer 1994; 73(8): 2013–2026
doi: 10.1002/1097-0142(19940415)73:8<2013::AID-CNCR2820730802>3.0.CO;2-J pmid: 8156506
3 Brinkworth MH, Weinbauer GF, Bergmann M, Nieschlag E. Apoptosis as a mechanism of germ cell loss in elderly men. Int J Androl 1997; 20(4): 222–228
doi: 10.1046/j.1365-2605.1997.00056.x pmid: 9401825
4 Dunkel L, Hirvonen V, Erkkil? K. Clinical aspects of male germ cell apoptosis during testis development and spermatogenesis. Cell Death Differ 1997; 4(3): 171–179
doi: 10.1038/sj.cdd.4400234 pmid: 16465226
5 Hikim AP, Wang C, Lue Y, Johnson L, Wang XH, Swerdloff RS. Spontaneous germ cell apoptosis in humans: evidence for ethnic differences in the susceptibility of germ cells to programmed cell death. J Clin Endocrinol Metab 1998; 83(1): 152–156
doi: 10.1210/jcem.83.1.4485 pmid: 9435433
6 Sinha Hikim AP, Swerdloff RS. Hormonal and genetic control of germ cell apoptosis in the testis. Rev Reprod 1999; 4(1): 38–47
doi: 10.1530/ror.0.0040038 pmid: 10051101
7 Opferman JT, Iwasaki H, Ong CC, Suh H, Mizuno S, Akashi K, Korsmeyer SJ. Obligate role of anti-apoptotic MCL-1 in the survival of hematopoietic stem cells. Science 2005; 307(5712): 1101–1104
doi: 10.1126/science.1106114 pmid: 15718471
8 Lipshultz LI, Howards SS. Evaluation of the subfertile male. In: Lipshultz LI, Howards SS. Infertility in the Male. 3rd ed. St. Louis: Mosby-Year Book, Inc., 1997: 173–193
9 Coburn M, Kim ED, Wheeler TM. Testicular biopsy in male infertility evaluation. In: Lipshultz LI, Howards SS. Infertility in the Male. 3rd ed. St. Louis: Mosby-Year Book, Inc., 1997: 219–248
10 Kovacs G. Assessment of the male partner. In: Kovacs G. The Subfertility Handbook: a Clinician’s Guide. Cambridge: Cambridge University Press, 2010: 54–59
11 Boenisch T, Farmilo AJ, Stead RH, Key M, Welcher R, Harvey R, Atwood KN. Hand Book Immunochemical Staining Methods, 3rd ed. Carpinteria, California USA: DAKO Corporation, 2001: 1–62
12 Kukner A, Oner J, Oner H, Ozan E. Effect of vitamin E on expression of Appoptosis Related Proteins in Immobilized Rat testes. J <?Pub Caret?>Anim Vet Adv 2010; 9(1): 180–185
doi: 10.3923/javaa.2010.180.185
13 Sugiyama N, Obinata M, Matsui Y. Bcl-2 inhibits apoptosis of spermatogonia and growth of spermatogonial stem cells in a cell-intrinsic manner. Mol Reprod Dev 2001; 58(1): 30–38
doi: 10.1002/1098-2795(200101)58:1<30::AID-MRD5>3.0.CO;2-4 pmid: 11144217
14 Rinkenberger JL, Horning S, Klocke B, Roth K, Korsmeyer SJ. Mcl-1 deficiency results in peri-implantation embryonic lethality. Genes Dev 2000; 14(1): 23–27
pmid: 10640272
15 Yang T, Buchan HL, Townsend KJ, Craig RW. MCL-1, a member of the BLC-2 family, is induced rapidly in response to signals for cell differentiation or death, but not to signals for cell proliferation. J Cell Physiol 1996; 166(3): 523–536
doi: 10.1002/(SICI)1097-4652(199603)166:3<523::AID-JCP7>3.0.CO;2-R pmid: 8600156
16 Oldereid NB, Angelis PD, Wiger R, Clausen OP. Expression of Bcl-2 family proteins and spontaneous apoptosis in normal human testis. Mol Hum Reprod 2001; 7(5): 403–408
doi: 10.1093/molehr/7.5.403 pmid: 11331661
17 El-Fackahany HM, Sakkas D. Abortive apoptosis and sperm chromatin damage. In: Zini A, Agarwell A. Sperm Chromatin: Biological and Clinical Applications in Male Infertility. New York: Springer-Verlag New York, 2011
[1] Yujiao Liu,Chao Liu,Wen Dong,Wei Li. Physiological functions and clinical implications of the N-end rule pathway[J]. Front. Med., 2016, 10(3): 258-270.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed