Mechanisms of calcium oscillations in mouse and human eggs

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Elgmati, Khalil;
  • Subject: QH301 | QH426

Long lasting calcium (Ca2+) oscillations are necessary and sufficient for mammalian egg activation and early embryological development. In mammals, phospholipase C zeta (PLCζ) has been identified as the likely endogenous trigger of Ca2+ oscillations at fertilization. So... View more
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    2.1 HEALTH AND SAFETY & LEGAL PROCEDURES ----------------------------------------------------------------------------- 88

    2.2 MATERIALS ---------------------------------------------------------------------------------------------------------------------- 88 2.2.1 Super-ovulation and egg collection -------------------------------------------------------------------------------- 89 2.2.1.1 Mouse eggs --------------------------------------------------------------------------------------------------------------------- 89 2.2.1.2 Human eggs -------------------------------------------------------------------------------------------------------------------- 89

    2.3 METHODS ------------------------------------------------------------------------------------------------------------------------ 91 2.3.1 Microinjection -------------------------------------------------------------------------------------------------------- 91 2.3.1.1 The injection procedure for PLCζ experiments---------------------------------------------------------------------------- 93 2.3.1.2 The injection procedure for Sr2+ experiments------------------------------------------------------------------------------ 94 2.3.1.3 The procedure for Sr2+ injection into mouse/human eggs ---------------------------------------------------------------- 94 2.3.2 Media preparation --------------------------------------------------------------------------------------------------- 96 2.3.2.1 H-KSOM (Hepes-buffered Potassium Simplex Optimised Media). ---------------------------------------------------- 96 2.3.2.2 Sr2+ H-KSOM Preparation---------------------------------------------------------------------------------------------------- 96

    3.1 INTRODUCTION---------------------------------------------------------------------------------------------------------------- 107 3.1.1 Critical levels of extracellular Sr2+ are required to trigger intracellular Ca2+-oscillations. --------------108 3.1.2 How does extracellular Sr2+ trigger Ca2+ release? -------------------------------------------------------------109 3.1.3 Objectives ------------------------------------------------------------------------------------------------------------110

    3.2 STRONTIUM INJECTION INTO MOUSE EGGS & CALCIUM RELEASE ----------------------------------------------------- 110 3.2.1 KCl injection into mouse eggs and Ca2+ release ----------------------------------------------------------------112

    3.3 STRONTIUM-TRIGGERED OSCILLATIONS; ARE THEY CALCIUM OR STRONTIUM TRANSIENTS? -------------------- 113 3.3.1 Sr2+-triggered fluorescence and ionomycin ----------------------------------------------------------------------113

    3.4 THE RELATIONSHIP BETWEEN STRONTIUM-TRIGGERED CALCIUM OSCILLATIONS AND MEDIUM OSMOLARITY 115 3.4.1 The effect of choline in low Na+ media on Sr2+-triggered Ca2+-oscillations --------------------------------120 3.4.2 The effect of high K+ in low Na+ media on Sr2+-triggered Ca2+-oscillations --------------------------------121 3.4.3 Na+ (0mM) media facilitates Sr2+-triggered Ca2+-oscillations in mouse eggs ------------------------------122 3.4.4 The relationship between Sr2+-triggered Ca2+-oscillations and mouse egg swelling-----------------------123

    3.6 DISCUSSION ------------------------------------------------------------------------------------------------------------------- 144 3.6.1 Sr2+ Influx can cause Ca2+-oscillations --------------------------------------------------------------------------144

    4.1 OVERVIEW OF PHOSPHOLIPASE CS ---------------------------------------------------------------------------------------- 150

    4.2 MALE INFERTILITY-LINKED POINT MUTATION THAT DISRUPTS THE CALCIUM OSCILLATION-INDUCING ACTIVITY

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