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Comparison of Ca2+ and CaMKII responses in IVF and ICSI in the mouse

Abstract Novel methods of egg activation in human assisted reproductive technologies and animal somatic cell nuclear transfer are likely to alter the signalling process that occurs during normal fertilization. Intracytoplasmic sperm injection (ICSI) bypasses the normal processes of the acrosome reaction, sperm–egg fusion, and processing of the sperm plasma membrane, as well as alters some parameters of intracellular calcium (Ca 2+ i ) dynamics (reported previously by Kurokawa and Fissore (2003)). Herein, we extend these studies to determine if ICSI alters the activity of the Ca 2+ -dependent protein, Ca 2+ /calmodulin-dependent kinase II (CaMKII), which is responsible for the completion of meiosis in vertebrate eggs. After ICSI or in vitro fertilization (IVF), individual mouse eggs were monitored for their relative changes in both Ca 2+ i and CaMKII activity during the first Ca 2+ i rise and a subsequent rise associated with second polar body extrusion. The duration of the first Ca 2+ i rise was greater in ICSI than in IVF, but the amplitude of the rise was transiently higher for IVF than ICSI. However, a similar mean CaMKII activity was observed in both procedures. During polar body extrusion, the amplitude and duration of the Ca 2+ rises were increased by a small amount in ICSI compared with IVF, whereas the CaMKII activities were similar. Thus, compared with IVF, ICSI is not associated with decreased or delayed CaMKII activity in response to these Ca 2+ signals in the mouse. Key words http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Molecular Human Reproduction Oxford University Press

Comparison of Ca2+ and CaMKII responses in IVF and ICSI in the mouse

Abstract

Abstract Novel methods of egg activation in human assisted reproductive technologies and animal somatic cell nuclear transfer are likely to alter the signalling process that occurs during normal fertilization. Intracytoplasmic sperm injection (ICSI) bypasses the normal processes of the acrosome reaction, sperm–egg fusion, and processing of the sperm plasma membrane, as well as alters some parameters of intracellular calcium (Ca 2+ i ) dynamics (reported previously by Kurokawa and Fissore (2003)). Herein, we extend these studies to determine if ICSI alters the activity of the Ca 2+ -dependent protein, Ca 2+ /calmodulin-dependent kinase II (CaMKII), which is responsible for the completion of meiosis in vertebrate eggs. After ICSI or in vitro fertilization (IVF), individual mouse eggs were monitored for their relative changes in both Ca 2+ i and CaMKII activity during the first Ca 2+ i rise and a subsequent rise associated with second polar body extrusion. The duration of the first Ca 2+ i rise was greater in ICSI than in IVF, but the amplitude of the rise was transiently higher for IVF than ICSI. However, a similar mean CaMKII activity was observed in both procedures. During polar body extrusion, the amplitude and duration of the Ca 2+ rises were increased by a small amount in ICSI compared with IVF, whereas the CaMKII activities were similar. Thus, compared with IVF, ICSI is not associated with decreased or delayed CaMKII activity in response to these Ca 2+ signals in the mouse. Key words
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