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. 2015;61(6):519-24.
doi: 10.1262/jrd.2015-073. Epub 2015 Aug 21.

Effects of acrosomal conditions of frozen-thawed spermatozoa on the results of artificial insemination in Japanese Black cattle

Kazumi Kishida  1 , Mitsuhiro Sakase, Kenta Minami, Miyuki M Arai, Reiko Syoji, Namiko Kohama, Takayuki Akiyama, Akio Oka, Hiroshi Harayama, Moriyuki Fukushima
Affiliations

Effects of acrosomal conditions of frozen-thawed spermatozoa on the results of artificial insemination in Japanese Black cattle

Kazumi Kishida et al. J Reprod Dev. 2015.

Abstract

The purposes of this study were to examine the relationship between male artificial insemination (AI) fertility and sperm acrosomal conditions assessed by new and conventional staining techniques and to identify possible reproductive dysfunctions causing low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions in Japanese Black bulls. We investigated individual differences among bulls in the results concerning (1) acrosomal conditions of frozen-thawed spermatozoa as assessed by not merely peanut agglutinin-lectin staining (a conventional staining technique) but also immunostaining of acrosomal tyrosine-phosphorylated proteins (a new staining technique), (2) routine AI using frozen-thawed spermatozoa as assessed by pregnancy diagnosis, (3) in vivo fertilization of frozen-thawed spermatozoa and early development of fertilized eggs as assessed by superovulation/AI-embryo collection tests and (4) in vitro fertilization of frozen-thawed spermatozoa with oocytes. The percentages of frozen-thawed spermatozoa with normal acrosomal conditions assessed by the abovementioned staining techniques were significantly correlated with the conception rates of routine AI, rates of transferable embryos in superovulation/AI-embryo collection tests and in vitro fertilization rates. These results are consistent with new suggestions that the distribution of acrosomal tyrosine-phosphorylated proteins as well as the acrosomal morphology of frozen-thawed spermatozoa are AI fertility-associated markers that are valid for the prediction of AI results and that low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions result from reproductive dysfunctions in the processes between sperm insemination into females and early embryo development, probably failed fertilization of frozen-thawed spermatozoa with oocytes.

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Figures

Fig. 1.
Fig. 1.
Relationship between acrosomal integrity and distributive normality of acrosomal tyrosine-phosphorylated (pY) proteins in frozen-thawed spermatozoa from Japanese Black bulls. Filled circles (●) indicate the results obtained for each of 34 bulls (A1–A9, B1–B9, C1–C9 and D1–D7). a Acrosomal integrity was examined by the fluorescein isothiocyanate (FITC)-conjugated peanut agglutinin (PNA)/propidium iodide (PI) staining. bDistributive normality of acrosomal pY proteins was examined by indirect immunofluorescence.
Fig. 2.
Fig. 2.
Relationship between acrosomal conditions of frozen-thawed spermatozoa and results of artificial insemination (AI) using frozen-thawed spermatozoa in Japanese Black bulls. Filled circles (●) indicate the results obtained for each of the bulls (left panel, 19 bulls, A2, A4–A9, B1–B9, D8, D9 and E1; right panel, 16 bulls A2, A4–A9 and B1–B9).
Fig. 3.
Fig. 3.
Relationship between acrosomal conditions of frozen-thawed spermatozoa and results of superovulation/artificial insemination-embryo collection (SOV/AI-EC) tests using frozen-thawed spermatozoa in Japanese Black bulls. Filled circles (●) indicate the results obtained for each of 16 bulls (A1, A3–A9, B1-B5 and C1–C3).
Fig. 4.
Fig. 4.
Relationship between rapidly progressive movement of frozen-thawed spermatozoa and results of SOV/AI-EC tests using frozen-thawed spermatozoa in Japanese Black bulls. Filled circles (●) indicate the results obtained for each of 16 bulls (A1, A3–A9, B1–B5 and C1–C3).
Fig. 5.
Fig. 5.
Results of in vitro fertilization (IVF) tests using frozen-thawed spermatozoa with different acrosomal conditions in Japanese Black bulls. Values are means ± SEM.
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