Sperm Na+, K+-ATPase and Ca2+-ATPase activity: A preliminary study of comparison of swim up and density gradient centrifugation methods for sperm preparation

Silvia W. Lestari, Manggiasih D. Larasati, Asmarinah, Indra G. Mansur

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

Abstract

As one of the treatment for infertility, the success rate of Intrauterine Insemination (IUI) is still relatively low. Several sperm preparation methods, swim-up (SU) and the density-gradient centrifugation (DGC) are frequently used to select for better sperm quality which also contribute to IUI failure. Sperm selection methods mainly separate the motile from the immotile sperm, eliminating the seminal plasma. The sperm motility involves the structure and function of sperm membrane in maintaining the balance of ion transport system which is regulated by the Na+, K+-ATPase, and Ca2+-ATPase enzymes. This study aims to re-evaluate the efficiency of these methods in selecting for sperm before being used for IUI and based the evaluation on sperm Na+,K+-ATPase and Ca2+-ATPase activities. Fourteen infertile men from couples who underwent IUI were involved in this study. The SU and DGC methods were used for the sperm preparation. Semen analysis was performed based on the reference value of World Health Organization (WHO) 2010. After isolating the membrane fraction of sperms, the Na+, K+-ATPase activity was defined as the difference in the released inorganic phosphate (Pi) with and without the existence of 10 mM ouabain in the reaction, while the Ca2+-ATPase was determined as the difference in Pi contents with and without the existence of 55 μm CaCl2. The prepared sperm demonstrated a higher percentage of motile sperm compared to sperm from the whole semen. Additionally, the percentage of motile sperm of post-DGC showed higher result than the sperm from post-SU. The velocity of sperm showed similar pattern with the percentage of motile sperm, in which the velocity of prepared sperm was higher than the sperm from whole semen. Furthermore, the sperm velocity of post-DGC was higher compared to the sperm from post-SU. The Na+, K+-ATPase activity of prepared sperm was higher compared to whole semen, whereas Na+, K+-ATPase activity in the post DGC was higher than post SU. The Ca2+-ATPase activity of prepared sperm was higher compared to whole semen, whereas Ca2+-ATPase activity in the post DGC was higher than post SU. The SU and the DGC methods were able to perform sperm selection by showing a high result of Na+, K+-ATPase and Ca2+-ATPase activities, moreover DGC method selected the sperm with high activities of both the Na+, K+-ATPase and Ca2+-ATPase better compared to SU method.

Original languageEnglish
Title of host publication2nd Biomedical Engineering�s Recent Progress in Biomaterials, Drugs Development, and Medical Devices
Subtitle of host publicationProceedings of the International Symposium of Biomedical Engineering, ISBE 2017
EditorsRadon Dhelika, Yudan Whulanza, Ghiska Ramahdita, Praswasti P.D.K. Wulan
PublisherAmerican Institute of Physics Inc.
ISBN (Electronic)9780735416253
DOIs
Publication statusPublished - 13 Feb 2018
Event2nd Biomedical Engineering's Recent Progress in Biomaterials, Drugs Development, and Medical Devices, ISBE 2017 - Bali, Indonesia
Duration: 25 Jul 201726 Jul 2017

Publication series

NameAIP Conference Proceedings
Volume1933
ISSN (Print)0094-243X
ISSN (Electronic)1551-7616

Conference

Conference2nd Biomedical Engineering's Recent Progress in Biomaterials, Drugs Development, and Medical Devices, ISBE 2017
Country/TerritoryIndonesia
CityBali
Period25/07/1726/07/17

Keywords

  • Ca-ATPase
  • K-ATPase
  • Na
  • density gradient centrifugation
  • swim-up

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