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Elimination of apoptotic spermatozoa from rabbit insemination dose using annexin V associated with the MACS
Jaromir Vasicek1, Vladimir Parkanyi, Lubomir Ondruska
1Animal Production Research Centre Nitra, Hlohovecká 2, 951 41 Luzianky, Slovak Republic. jaromir.vasicek@gmail.com
Abstract:
The aim of this study was to verify whether the separation and elimination of the apoptotic fraction in rabbit semen using a MACS technique may improve sperm fertility potential and consequently rabbit kindling rate. Semen samples from 25 New Zealand White (NZW) rabbit males were collected using an artificial vagina and evaluated using the CASA system for concentration and motility. For artificial insemination the best 11 bucks were chosen based on motility parameters. Their ejaculates were mixed to make a heterospermic pool and routinely diluted in a commercial insemination diluent (MiniTüb, Tiefenbach, Germany) at a ratio of 1:6. Diluted heterospermic spermatozoa were filtered through a Sartorius filter to wash out seminal plasma, re-diluted in binding buffer (Annexin V Microbead Kit, Miltenyi Biotec, Germany) at a ratio of 1:3.66 and divided into two groups: an experimental group intended for MACS separation and control group without MACS separation. Then hormonally treated females of NZW rabbits were inseminated with fresh doses of filtered heterospermic semen (n = 27; 0.5 ml I.D. per female) and MACS separated semen (n=28; 0.5 ml I.D. per female). Separation and subsequent elimination of apoptotic spermatozoa (positive selection) from the insemination dose (after negative MACS selection) was verified under in vivo conditions on the basis of increased kindling rate in the experimental group in comparison with kindling rate in the control group (81.3% vs. 73.8%). In conclusion, elimination of apoptotic spermatozoa by the use of the MACS technique results in a slight improvement in kindling rate of rabbit does.
Insights
Eliminating apoptotic spermatozoa using Magnetic-Activated Cell Sorting (MACS) slightly improved rabbit kindling rates. This technique enhances sperm fertility potential in artificial insemination for New Zealand White rabbits.
Area of Science:
- Animal Reproduction
- Sperm Biology
- Biotechnology
Background:
- Apoptotic spermatozoa can negatively impact fertility.
- Improving artificial insemination efficiency is crucial in rabbit breeding.
- Magnetic-Activated Cell Sorting (MACS) offers a method for sperm selection.
Purpose of the Study:
- To determine if removing apoptotic sperm via MACS enhances rabbit fertility.
- To assess the effect of MACS-sorted semen on kindling rates in New Zealand White rabbits.
Main Methods:
- Semen from New Zealand White rabbits was collected and evaluated.
- Apoptotic spermatozoa were separated using MACS.
- Hormonally treated does were inseminated with MACS-treated or untreated semen.
- Kindling rates were compared between groups.
Main Results:
- The group inseminated with MACS-separated semen showed a kindling rate of 81.3%.
- The control group (untreated semen) had a kindling rate of 73.8%.
- A slight increase in kindling rate was observed in the MACS-treated group.
Conclusions:
- Elimination of apoptotic spermatozoa using MACS leads to a modest improvement in rabbit does' kindling rate.
- MACS technique shows potential for enhancing sperm fertility and reproductive efficiency in rabbits.
