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      Exogenous neurotensin modulates sperm function in Japanese Black cattle

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          Abstract

          Recently, the conception rates after artificial insemination have been pointed out to decline continuously. To overcome this problem, the control of frozen and thawed sperm quality is required. However, the mechanism of bovine sperm functional regulation is still largely unknown. In mammals, the ejaculated sperm are capable of showing fertilizing ability during migration in the female reproductive organs. It is well known that these female organs secrete several factors contributing to sperm capacitation. We previously reported that neurotensin (NT) secreted from the oviduct and cumulus cells enhanced sperm capacitation and acrosome reaction in mice. In this study, we confirmed the expression of the NT receptor (NTR1) in the bovine sperm neck region and the secretion of NT in the bovine uterus and oviduct. The similar expression patterns of NT and NTR1 suggests a conserved mechanism of sperm functional regulation between mouse and cattle. Thus, we examined the effects of exogenous NT on the bovine sperm functions. First, we showed that NT induced sperm protein tyrosine phosphorylation in a dose-dependent manner, suggesting that NT enhances sperm capacitation. Second, we showed that NT induced acrosome reactions of capacitated sperm in a dose-dependent manner, suggesting that NT facilitates acrosome reaction. Finally, we used a computer-aided sperm analysis system to show that NT did not have a great effect on sperm motility. These results suggest that NT acts as a facilitator of sperm capacitation and acrosome reaction in the female reproductive tracts in cattle, highlighting the importance of NT-mediated signaling to regulate sperm functions.

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          Most cited references34

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          Capacitation of bovine sperm by heparin.

          Capacitation of bovine sperm was evaluated by determining the ability of sperm to fertilize bovine oocytes in vitro and to undergo an acrosome reaction upon exposure to lysophosphatidylcholine (LC). Incubation of sperm with heparin (10 micrograms/ml) increased the percentage of oocytes fertilized, but this required exposing sperm to heparin for at least 4 h before adding them to oocytes. There was no effect on the percentage of motile or acrosome-reacted sperm after exposure of noncapacitated sperm to 100 micrograms/ml LC for 15 min. When sperm were incubated for 4 h with heparin, exposure to 100 micrograms/ml LC for 15 min had no effect on the percentage of sperm that were motile, but the percentage of acrosome-reacted sperm increased from less than 10% to over 70%. The acrosome reactions (ARs) induced by LC were synchronous, reached maximal levels within 15 min, and differed (p less than 0.001) between sperm incubated under capacitating (with heparin) and noncapacitating conditions (without heparin). The time course required for heparin to capacitate sperm as judged by in vitro fertilization and to render sperm sensitive to LC induction of the AR were found to be similar. The percentage of ARs induced by LC and percentage of oocytes fertilized by sperm were found to be heparin-dose-dependent, with the maximum responses occurring at 5-10 micrograms/ml heparin. The correlation between the mean fertilization and LC-induced AR percentages was 0.997 (p less than 0.01). These studies demonstrate capacitation of bovine sperm by heparin requires at least a 4-h exposure of sperm to heparin and suggest that plasma membrane changes prior to an AR can be detected by exposure of bovine sperm to LC.
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            The isolation of a new hypotensive peptide, neurotensin, from bovine hypothalami.

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              Basic aspects of frozen storage of semen.

              W.V. HOLT (2000)
              Basic concepts of cryopreservation and the causes of cryoinjury are reviewed. The possible roles of cryoprotectants and additives are considered in the context of their putative interactions with the sperm plasma membrane. Modern approaches to the laboratory assessment of spermatozoa after freeze-thawing are also briefly discussed.
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                Author and article information

                Journal
                J Reprod Dev
                J. Reprod. Dev
                JRD
                The Journal of Reproduction and Development
                The Society for Reproduction and Development
                0916-8818
                1348-4400
                20 May 2016
                August 2016
                : 62
                : 4
                : 409-414
                Affiliations
                [1) ]Laboratory of Animal Reproduction and Development, Graduate School of Agricultural Science, Tohoku University, Miyagi 981-8555, Japan
                [2) ]Miyagi Prefectural Livestock Experiment Station, Miyagi 989-6445, Japan
                [3) ]Miyagi Agricultural Development Corporation, Miyagi 981-0914, Japan
                Author notes
                Correspondence: K Tanemura (e-mail: kentaro@ 123456m.tohoku.ac.jp )
                Article
                2016-055
                10.1262/jrd.2016-055
                5004797
                27210588
                e42aa723-31f8-43a2-96db-a99973507622
                ©2016 Society for Reproduction and Development

                This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License.

                History
                : 04 April 2016
                : 21 April 2016
                Categories
                Original Article

                acrosome reaction,bovine sperm,capacitation,neurotensin,sperm motility

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