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Integrins are not involved in the process of human sperm-oolemmal fusion
Authors:Sengoku Kazuo  Takuma Naoyuki  Miyamoto Toshinobu  Horikawa Michiharu  Ishikawa Mutsuo
Affiliation:Department of Obstetrics and Gynecology, Asahikawa Medical College, Midorigaoka-higashi 2-1, Asahikawa, 0788510, Japan
Abstract:BACKGROUND: We investigated whether integrins are required forthe human sperm–oocyte binding and fusion processes. METHODS:The expression of several integrin subunits at the human oocyteplasma membrane was investigated using immunofluorescence microscopy,and the functional role of integrins expressed at the humanoocyte surface in sperm–oocyte interaction was studiedusing a zona-free human oocyte binding and fusion assay. A totalof 144 unfertilized oocytes were stained with anti-integrinantibodies and 147 zona-free unfertilized oocytes were inseminatedin the presence of various anti-integrin antibodies that wereexpressed in oocyte plasma membrane. RESULTS: The antibodiesof six {alpha} integrin subunits ({alpha}2, {alpha}3, {alpha}5, {alpha}6, {alpha}V, {alpha}M) and six {beta} integrinsubunits ({beta}1, {beta}2, {beta}3, {beta}4, {beta}5, {beta}6) were bound to the surface of fixedunfertilized oocytes. In contrast, the presence of {alpha}1 and {alpha}4 subunitscould not be verified. The human sperm–oocyte bindingwas only partially inhibited by blocking antibodies of {alpha}2, {alpha}3,{alpha}5, {alpha}6, {alpha}V, {alpha}M, {beta}1, {beta}2 and {beta}3 with a maximum of 55% inhibition, butantibodies of {beta}4, {beta}5 and {beta}6 showed no effect on sperm–oolemmalbinding. A similar reduction of the number of fused sperm wasobserved. However, the ratio of fused sperm to total sperm (boundand fused) was not impaired by all integrin antibodies, suggestingthat integrins had no role in the sperm–oolemmal fusionprocess. CONCLUSIONS: These results suggest that one of thebinding mechanisms can be inhibited by integrin antibodies butthat this mechanism does not play an essential role in the humansperm–oolemmal binding and fusion processes. The othermechanisms, insensitive to integrins, may involve both bindingand fusion processes in human oocytes.
Keywords:human gamete fusion/integrin/oocyte plasma membrane/sperm
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