Reduction of truncated Kit Expression in Men with Abnormal Semen Parameters, Globozoospermia and History of Low or Fertilization Failure


Somayeh Haghighat, M.Sc, 1,2,#Marziyeh Tavalaee, Ph.D, 2,#Zahra Zakeri, Ph.D, 3Mahdi Noureddini, Ph.D, 1,*Abdolhossein Shahverdi, Ph.D, 4Mohammad Hossein Nasr Esfahani, Ph.D, 2,5,*
Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran
Department of Reproductive Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran
Department of Biology, Queens College and Graduate Center of the City University of New York, Flushing, NY, USA
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Medicine, ACECR, Tehran, Iran
Isfahan Fertility and Infertility Center, Isfahan, Iran
Physiology Research Center, Kashan University of Medical Sciences, Kashan, Iran
Department of Reproductive Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran
Department of Biology, Queens College and Graduate Center of the City University of New York, Flushing, NY, USA
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Medicine, ACECR, Tehran, Iran
Isfahan Fertility and Infertility Center, Isfahan, Iran
*Corresponding Addresses: P.O.Box: 87155/111 Physiology Research Center Kashan University of Medical Sciences Kashan Iran P.O.Box: 8165131378 Department of Reproductive Biotechnology Reproductive Biomedicine Research Center Royan Institute for Biotechnology ACECR Isfahan Iran Emails:mnoureddini@yahoo.com,mh.nasr-esfahani@royaninstitute.org

The first two authors equally contributed to this work.

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Haghighat Somayeh, Tavalaee Marziyeh, Zakeri Zahra, Noureddini Mahdi, Shahverdi Abdolhossein, Nasr Esfahani Mohammad Hossein. Reduction of truncated Kit Expression in Men with Abnormal Semen Parameters, Globozoospermia and History of Low or Fertilization Failure. Cell J. 2019; 21(3): 314-321.

Abstract

Objective

Phospholipase C zeta 1 (PLCζ) is one of the main sperm factor involved in oocyte activation and other factors may assist this factor to induce successful fertilization. Microinjection of recombinant tr-kit, a truncated form of c-kit receptor, into metaphase II-arrested mouse oocytes initiate egg activation. Considering the potential roles of tr- KIT during spermiogenesis and fertilization, we aimed to assess expression of tr-KIT in sperm of men with normal and abnormal parameters and also in infertile men with previous failed fertilization and globozoospermia.

Materials and Methods

This experimental study was conducted from September 2015 to July 2016 on 30 normozoospermic and 20 abnormozoospermic samples for experiment one, and also was carried out on 10 globozoospermic men, 10 men with a history low or failed fertilization and 13 fertile men for experiment two. Semen parameters and sperm DNA fragmentation were assessed according to WHO protocol, and TUNEL assay. Sperm tr- KIT was evaluated by flow cytometry, immunostaining and western blot.

Results

The results show that tr-KIT mainly was detected in post-acrosomal, equatorial and tail regions. Percentage of tr-KIT-positive spermatozoa in abnormozoospermic men was significantly lower than normozoospermic men. Also significant correlations were observed between sperm tr-KIT with sperm count (r=0.8, P<0.001), motility (r=0.31, P=0.03) and abnormal morphology (r=-0.6, P<0.001). Expression of tr-KIT protein was significantly lower in infertile men with low/ failed fertilization and globozoospermia compared to fertile men. The significant correlation was also observed between tr-KIT protein with fertilization rate (r=-0.46, P=0.04). In addition, significant correlations were observed between sperm DNA fragmentation with fertilization rate (r=-0.56, P=0.019) and tr-KIT protein (r=-0.38, P=0.04).

Conclusion

tr-KIT may play a direct or indirect role in fertilization. Therefore, to increase our insight regarding the role of tr-KIT in fertilization further research is warranted.