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Selected Publication:

Publication type: Journal Article
Document type: Full Paper

Year: 2000

Author(s): Suttner, R; Zakhartchenko, V; Stojkovic, P; Müller, S; Alberio, R; Medjugorac, I; Brem, G; Wolf, E; Stojkovic, M

Title: Intracytoplasmic sperm injection in bovine: effects of oocyte activation, sperm pretreatment and injection technique.

Source: Theriogenology. 2000; 54(6):935-948



Authors Vetmeduni Vienna:

Brem Gottfried
Müller Simone


Abstract:
Intracytoplasmic sperm injection (ICSI) is a very important technique for treating male subfertility and for basic research. The efficiency of ICSI in bovine is very limited because of the necessity for additional oocyte activation before or after the ICSI procedure. In this study, we compared the effects of seven different protocols on activation and fertilization rates of bovine oocytes after ICSI and on their subsequent development under in vitro conditions. The protocols include 1) different chemical activation of oocytes, 2) pretreated or nonpretreated sperm, and 3) conventional or Piezo-driven injection techniques. In all three groups, ICSI, sham-injected, and noninjected, the highest activation rates were obtained after treatment of oocytes with ionomycin followed by 6-dimethylaminopurine (6-DMAP). Using this treatment for oocyte activation, 59% of oocytes were activated and 31% of oocytes were fertilized using dithiothreitol (DTT) pretreated spermatozoa and Piezo-driven injection. Using the protocols with the same oocyte activation or activation with calcium ionophore (Ca-I) and cycloheximide (CHX), nonpretreated sperm, and conventional injection technique, early cleavage rate (79.6% and 77.6%, respectively) were significantly (P <0.01) higher when compared with all other protocols. The latter protocol resulted in 8% blastocyst and 90% of the obtained blastocysts were found to be diploid. Our results demonstrate that activation of oocytes, sperm treatment, and injection technique separately or together could improve the success of bovine ICSI.

Keywords Pubmed: Adenine/analogs & derivatives
Adenine/pharmacology
Animals
Benzimidazoles/chemistry
Calcimycin/pharmacology
Cattle/physiology*
Coloring Agents/chemistry
Cycloheximide/pharmacology
DNA/chemistry
DNA/isolation & purification
DNA Primers/chemistry
Enzyme Inhibitors/pharmacology
Female
Fluorescent Dyes/chemistry
Ionomycin/pharmacology
Ionophores/pharmacology
Karyotyping
Least-Squares Analysis
Male
Oocytes/physiology*
Oxazines/chemistry
Ploidies
Polymerase Chain Reaction/veterinary
Polymorphism, Restriction Fragment Length
Sex Determination Analysis/veterinary
Sperm Injections, Intracytoplasmic/methods
Sperm Injections, Intracytoplasmic/veterinary*
Sperm-Ovum Interactions/physiology*
Spermatozoa/physiology*


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