Duru ARAS TOSUN, Elmas Yaren SUİÇMEZ, Derya GÖKMEN, Sinan ÖZKAVUKÇU

Farklı Likefaksiyon Sıcaklıklarının Semen Ozmolalitesi ve Sperm Canlılığı Üzerine Etkileri

Semen likefaksiyonu, jel görünümlü semenin prostatta üretilen proteazların enzimatik aktivitesi ile sıvılaştığı proteolitik bir işlemdir. Semen likefaksiyonu için üremeye yardımcı tedavi laboratuvarları arasında farklı uygulamalar benimsenmiştir. Farklı sıcaklıkların semen ozmolalitesinde yol açtığı değişikliklerin sperm canlılığı üzerindeki etkileri bilinmemektedir. Bu metodolojik çalışmanın amacı, semen likefaksiyonu farklı sıcaklıklarda gerçekleştirildiğinde semen ozmolalitesinde meydana gelebilecek olası değişimlerin sperm canlılığına etkilerini ortaya koymaktır. Çalışmaya bir Üreme Yardımcı Tedavi Merkezi'ne semen analizi veya intrauterin inseminasyon için başvuran erkek hastalar dahil edilmiştir. Toplam 15 hastadan alınan semen örnekleri iki gruba ayrılarak 37°C'de veya oda sıcaklığında inkübe edilmiştir. Her bir örnek için ozmolalite ölçümü likefaksiyonu takiben donma noktası depresyon ozmometresi ile yapılmıştır. Sperm hareketliliği Makler sayım kamarasıyla belirlenmiş ve hareket tipleri Dünya Sağlık Örgütü tarafından yayınlanan Semen Analizi Kılavuzu’nda belirtildiği şekilde sınıflandırılmıştır. Sperm canlılığı eozin-nigrozin boyamasıyla test edilmiştir. Gruplar arasındaki fark student’s t testiyle belirlenmiştir. Sonuçlar p

Anahtar Kelimeler:

Likefaksiyon, Sıcaklık, Semen ozmolalitesi, Sperm canlılığı

The Effects of Different Liquefaction Temperatures on Semen Osmolality and Sperm Viability

Semen liquefaction is a proteolytic process in which semen is liquefied by the enzymatic activity of proteases derived from the prostate. Different practices are adopted among assisted reproduction laboratories for semen liquefaction. Effects of different temperatures to sperm viability due to osmotic changes are unknown. The aim of this methodological study is to reveal the changes in semen osmolality and sperm viability when liquefied at different temperatures. Male patients who applied to an Assisted Reproduction Technologies Center for semen analysis or intrauterine insemination were included to the study. Semen samples from 15 individuals were divided into two groups before liquefaction and were incubated at 37°C or room temperature. For each sample, osmolality measurement was performed with freezing point depression osmometer. Sperm motility was determined by the Makler counting chamber and motility types were classified as specified in the Guideline for Semen Analysis published by the World Health Organization. Sperm viability was determined by eosin-nigrosine staining following liquefaction. Student’s t-test was utilized to compare groups and the significance level was set at p

Keywords:

Liquefaction, Temperature, Semen osmolality, Sperm viability,

PDF

___

1. Reinhardt K, Dobler R, Abbott J. An ecology of sperm: sperm diversification by natural selection. Annu Rev Ecol Evol Syst. 2015;46:435-459.
2. Lupold S, Pitnick S. Sperm form and function: What do we know about the role of sexual selection? Reproduction. 2018;155(5):R229-R243.
3. Kayman-Kurekci G, Bunsuz M, Önal G, Dincer P. Kazanılmış epigenetik değişikliklerin kalıtımı ve hastalıklara yatkınlıktaki rolü. J Ist Faculty Med. 2017; 80:1:45-53
4. Lavanya M., Selvaraju S, Krishnappa B, ve ark. Microenvironment of the male and female reproductive tracts regulate sperm fertility: Impact of viscosity, pH, and osmolality. Andrology. 2022;10:92-104.
5. Katz D, Bloom T, Bondurant R. Movement of bull spermatozoa in cervical mucus. Biol Reprod. 1981;25(5):931-937.
6. Ho H-C, Suarez SS. Hyperactivation of mammalian spermatozoa: function and regulation. Reproduction. 2001;122(4):519-526.
7. Zhang Z, Ferraris JD, Brooks HL, Brisc I, Burg MB. Expression of osmotic stress-related genes in tissues of normal and hyposmotic rats. Am J Physiol Renal Physiol. 2003;285(4):F688-F693.
8. Cooper, TG, Yeung CH. Acquisition of volume regulatory response of sperm upon maturation in the epididymis and the role of the cytoplasmic droplet. Micros Res Tech 2003;61(1):28-38.
9. Björndahl L, Kvist U. Sequence of ejaculation affects the sper‐ matozoon as a carrier and its message. Reprod BioMed Online. 2003;7(4):440-448.
10. Holmes E, Björndahl L, Kvist U. Possible factors influencing post‐ejaculatory changes of the osmolality of human semen in vitro. Andrologia. 2019;51:e13443.
11. Anamthathmakula P, Winuthayanon W. Mechanism of semen liquefaction and its potential for a novel non-hormonal contraception. Biol Reprod 2020;103(2):411–426.
12. World Health Organization. WHO laboratory manual for the examination and processing of human semen. 6th edition. Geneva: World Health Organization; 2021.
13. Holmes E, Björndahl L, Kvist U. Post‐ejaculatory increase in human semen osmolality in vitro. Andrologia. 2019;51:e13311.
14. Nematollahi S, Mehdizadeh M, Hosseini S, ve ark. DNA integrity and methylation changes of mouse spermatozoa following prolonged incubation. Andrologia. 2019;51(6):e13276.
15. Matsuura R, Takeuchi T, Yoshida A. Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa. Asian J Androl. 2010;12(5):753-759.
16. Ameen, EM. Relationships of osmolality and oxidative stress with semen quality and their effects on male fertility. IHJPAS. 2021;IHICPAS:1-9.
17. Cooper TG, Barfield JP and Yeung CH. Changes in osmolality during liquefaction of human semen. Int J Androl. 2005;28:58-60.
18. Emami N, Deperthes D, Malm J, Diamandis EP. Major role of human KLK14 in seminal clot liquefaction. J Biol Chem. 2008;283(28):19561-19569.