CaMKIIα interacts with multi-PDZ domain protein MUPP1 in spermatozoa and prevents spontaneous acrosomal exocytosis

Ackermann, Frauke; Zitranski, Nele; Borth, Heike; Buech, Thomas; Gudermann, Thomas; Boekhoff, Ingrid

doi:10.1242/jcs.058263

Cited by 38 publications

(29 citation statements)

References 106 publications

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“…Streptolysin O (SLO) permeabilization is a widespread methodology used in analysis of dense-core granule exocytosis that we and others have applied to studying the AR (28,(36)(37)(38)(39). This method permits the introduction of exogenous factors-which are not otherwise able to cross membranes-into the cytosol through pores in the plasma membrane generated by SLO.…”

Section: Resultsmentioning

confidence: 99%

Rab27 and Rab3 sequentially regulate human sperm dense-core granule exocytosis

Bustos

Lucchesi

Ruete

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Two so-called “secretory Rabs,” Rab3 and Rab27, regulate late steps during dense-core vesicle exocytosis in neuroendocrine cells. Sperm contain a single large dense-core granule that is released by regulated exocytosis (termed the acrosome reaction) during fertilization or on exposure to inducers in vitro. Sperm exocytosis uses the same fusion machinery as neurons and neuroendocrine cells, with an additional requirement for active Rab3. Here we show that Rab27 is also required for the acrosome reaction, as demonstrated by the inability of inducers to elicit exocytosis when streptolysin O-permeabilized human sperm were loaded with inhibitory anti-Rab27 antibodies or the Rab27–GTP binding domain of the effector Slac2-b. The levels of GTP-bound Rab27 increased on initiation of exocytosis, as did the proportion of GTP-bound Rab3A. We have developed a fluorescence microscopy-based method for detecting endogenous Rab3A-GTP and Rab27-GTP in the acrosomal region of human sperm. Challenge with an inducer increased the population of cells exhibiting GTP-bound Rabs in this subcellular domain. Interestingly, introducing recombinant Rab27A loaded with GTP-γ-S into sperm elicited a remarkable increase in the number of cells evincing GTP-bound Rab3A. In the converse condition, recombinant Rab3A did not modify the percentage of Rab27-GTP–containing cells. Furthermore, Rab27A-GTP recruited a Rab3 GDP/GTP exchange factor (GEF) activity. Our findings suggest that Rab27/Rab3A constitutes a Rab-GEF cascade in dense-core vesicle exocytosis.

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Section: Resultsmentioning

confidence: 99%

Rab27 and Rab3 sequentially regulate human sperm dense-core granule exocytosis

Bustos

Lucchesi

Ruete

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Chelerythrine, U73122, U73343, Ro-31-7549, adenophostin A, 2-aminoethoxydiphenylborate (2-APB), xestospongin C, isopropyl ␤-D-1-thiogalactopyranoside, glutathione, FITC-conjugated Pisum sativum agglutinin (FITC-PSA), and EGTA were from Merck; glutathione-Sepharose 4B was from GE Healthcare. Phorbol 12-myristate 13-acetate (PMA), 1,2-dioctanoyl-sn-glycerol (DAG) (catalogue number D5156), phosphatidic acid (PA), FIPI hydrochloride hydrate, 1,2-bis(2-aminophenoxy)ethane-N,N,NЈ,NЈ-tetraacetic acid tetrakis (acetoxymethyl ester) (BAPTA-AM), and A23187 were from Sigma; phosphatidylinositol 4,5-bisphosphate (PIP 2 ) and phosphatidylinositol 4-phosphate (PI4P) were from Avanti Polar Lipids, Inc. (Alabaster, AL); EGTA-AM and Fluo3-AM were from Invitrogen; [␥- 32 P]ATP and Chemiluminescence Reagent Plus were from PerkinElmer Life Sciences and Migliore Laclaustra SRL (Buenos Aires, Argentina); 1-butanol was from BDH Chemicals; anti-Rab3A was from Synaptic Systems (mouse monoclonal antibody, purified IgG clone 42.2); anti-ARF6 (mouse monoclonal anti-human ARF6, ab19871 or ab131261) and anti-PLD1 (rabbit polyclonal anti-human PLD1, ab10585) were from Abcam (Cambridge, UK); anti-SNAP25 (monoclonal mouse antibody, clone 71.1; conserved epitope), anti-RIM, and anti-complexin I/II were from Synaptic Systems; Cy-3-conjugated anti-rabbit and HRP goat-conjugated antimouse were from Jackson ImmunoResearch. The rabbit polyclonal anti-GST antibody (purified IgG) was purchased from…”

Section: Methodsmentioning

confidence: 99%

“…Controlled plasma membrane permeabilization has been used to gain access to the membrane fusion machinery required for exocytosis in several secretory cells (29,30). We used a well established streptolysin O (SLO) permeabilization protocol for studying acrosomal granule exocytosis (31)(32)(33)(34) that allows the incorporation of exogenous proteins, lipids, and ions into the cytosol. The extracellular calcium acts as an exocytosis inducer mimicking the opening of store-operated calcium channels.…”

mentioning

confidence: 99%

ADP Ribosylation Factor 6 (ARF6) Promotes Acrosomal Exocytosis by Modulating Lipid Turnover and Rab3A Activation

Pelletán

Suhaiman

Vaquer

et al. 2015

Journal of Biological Chemistry

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Background: Sperm acrosomal exocytosis requires GTPases, SNAREs, and a complex lipid signaling. Results: Exocytic stimuli promote ARF6 activation, which accomplishes exocytosis by stimulating PLC and Rab3A. Conclusion: ARF6 induces acrosome calcium efflux and assembles the fusion machinery leading to membrane fusion. Significance: This study explores a novel molecular link between ARF6, PLC, and Rab3A and provides insight into the molecular mechanisms of exocytosis and reproduction.

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“…For the execution of the acrosome fusions additional Ca 2+ entry (in vitro by use of Ca 2+ ionophores, in vivo after zona binding) is required [9]. Diverse groups have shown that SNARE complex interacting proteins such as complexins [9,59, 60], dynamins [61], Rab 3A [62], synaptotagmins [63], multi-PDZ domain protein MUPP1, Calmodulin and CaMKIIalpha [64,65], Rab-2a, syntaxin binding proteins and Munc-18 (Tsai et al, unpublished results) have been discovered in sperm (see Fig. 5.2).…”

Section: Acrosome Exocytosismentioning

confidence: 99%

“…This is the site where the sperm binds to the zona pellucida and where the acrosome exocytosis as a response of that binding is inititated [46]. In that area within the aggregating lipid rafts MUPP1/CaMKIIα have been reported to interact with the trans ternary SNARE protein complex and this association functions as a fusion clamp [64,65]. (ii) The important factor in mouse sperm is the phosphorylated form of synaptotagmin, which appears to be important for preventing the acrosome exocytosis [63].…”

Section: Fig 52 Two Step Model For Snare Mediated Acrosome Exocytosmentioning

confidence: 99%

Membrane Fusions During Mammalian Fertilization

Gadella

Evans

2011

Advances in Experimental Medicine and Biology

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Successful completion of fertilization in mammals requires three different types of membrane fusion events. Firstly, the sperm cell will need to secrete its acrosome contents (acrosome exocytosis; also known as the acrosome reaction); this allows the sperm to penetrate the extracellular matrix of the oocyte (zona pellucida) and to reach the oocyte plasma membrane, the site of fertilization. Next the sperm cell will bind and fuse with the oocyte plasma membrane (also known as the oolemma), which is a different type of fusion in which two different cells fuse together. Finally, the fertilized oocyte needs to prevent polyspermic fertilization, or fertilization by more than one sperm. To this end, the oocyte secretes the contents of cortical granules by exocytotic fusions of these vesicles with the oocyte plasma membrane over the entire oocyte cell surface (also known as the cortical reaction or cortical granule exocytosis). The secreted cortical contents modify the zona pellucida, converting it to a state that is unreceptive to sperm, constituting a block to polyspermy. In addition, there is a block at the level of the oolemma (also known as the membrane block to polyspermy). IntroductionFertilization of the oocyte involves three membrane fusion events [1] namely, (1) a preparative series of secretion membrane fusions at the apical sperm surface known as acrosome exocytosis [2]. The membrane fusions are induced when the sperm cell binds to specific zona binding proteins at the sperm surface [3][4][5][6][7]. The acrosome exocytosis is a multipoint membrane fusion event between the sperm plasma membrane and the outer acrosomal membrane (see Fig. 5.1 [8,9]) and the exposed acrosomal content is required for sperm to penetrate the zona pellucida [10][11][12]. This so-called zona drilling effectively takes place because the sperm at this stage also has acquired hyperactivated motility [13]. (2) After zona penetration the sperm enters the perivitelline space where it can bind and fuse with the oocyte plasma membrane [14,15]. This is the actual fertilization fusion in which the contents of the sperm are delivered into the oocyte cytoplasm. The plasma membrane of the equatorial segment (see Fig. 5.1) is the site where proteins are located that orchestrate sperm-oocyte binding and fusion [16]. (3) In order to prevent polyspermy the oocyte has to activate defense systems to block redundant sperm-oocyte fusion [17]. To this end the first fertilizing sperm delivers activation factors

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CaMKIIα interacts with multi-PDZ domain protein MUPP1 in spermatozoa and prevents spontaneous acrosomal exocytosis

Cited by 38 publications

References 106 publications

Rab27 and Rab3 sequentially regulate human sperm dense-core granule exocytosis

Rab27 and Rab3 sequentially regulate human sperm dense-core granule exocytosis

ADP Ribosylation Factor 6 (ARF6) Promotes Acrosomal Exocytosis by Modulating Lipid Turnover and Rab3A Activation

Membrane Fusions During Mammalian Fertilization

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