VAMPs sensitive to tetanus toxin are required for cortical granule exocytosis in mouse oocytes

Paola, Matilde de; Garrido, Facundo; Zanetti, M. Natalia; Michaut, Marcela Alejandra

doi:10.1016/j.yexcr.2021.112629

Cited by 4 publications

(13 citation statements)

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“…In addition, the catalytic activity of the light chain is zinc-dependent and is used as a tool for the study of exocytosis in different secretory cells ( Ahnert-Hilger et al, 1989 ; Bittner et al, 1989 ). Previously we have demonstrated that VAMP1 and VAMP3 are expressed in mouse oocytes ( Paola et al, 2021 ). Considering that FAM-CPP internalization is innocuous for mouse oocytes ( Figure 2A ) and does not perturb cortical granules localization ( Figure 8 ), we hypothesized that a permeable tetanus Toxin light chain (CPP-TeTx) might inhibit cortical reaction by cleaving the VAMPs isoforms expressed in mouse oocytes.…”

Section: Resultsmentioning

confidence: 97%

“…Cortical reaction is a calcium regulated exocytosis, in which cortical granules fuse with the oocyte plasma membrane after mouse oocyte activation. This fusion is mediated by SNARE proteins, and we have proposed a working model for cortical granules exocytosis in mouse oocytes ( Paola et al, 2021 ). The SNARE proteins are proteolyzed by the light chains of the clostridial neurotoxins–tetanus toxin and botulinum toxin ( Humeau et al, 2000 ; Schiavo et al, 2000 ).…”

Section: Resultsmentioning

confidence: 99%

“…Our laboratory has made significant contributions to the identification of several proteins involved in this exocytosis process. We have demonstrated the participation of alpha-SNAP and NSF ( Paola et al, 2015 ), Rab3A ( Bello et al, 2016 ), as well as VAMP1 and VAMP3 ( Paola et al, 2021 ). Furthermore, other groups have identified SNAP23 ( Mehlmann et al, 2019 ), Rabphilin 3A ( Masumoto et al, 1996 ), Rab27A ( Wang et al, 2016 ), and Synaptotagmin I ( Zhu et al, 2020 ).…”

Section: Introductionmentioning

confidence: 95%

“…Furthermore, other groups have identified SNAP23 ( Mehlmann et al, 2019 ), Rabphilin 3A ( Masumoto et al, 1996 ), Rab27A ( Wang et al, 2016 ), and Synaptotagmin I ( Zhu et al, 2020 ). Collectively, these findings have allowed us to propose a comprehensive working model for membrane fusion during cortical granule exocytosis in mouse oocytes ( Paola et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

“…To identify the participation of a specific protein in the cortical reaction, we developed a functional assay based on cortical granules quantification during the cortical reaction parthenogenetically activated in mouse oocytes. To inhibit protein function, we performed microinjections of recombinant proteins, blocking antibodies, or toxins prior to parthenogenetic activation with strontium chloride ( Paola et al, 2015 ; Bello et al, 2016 ; Paola et al, 2021 ). So far, oocyte microinjection is the standard method for introducing molecules into oocytes.…”

Section: Introductionmentioning

confidence: 99%

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The light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibits cortical reaction in mouse oocytes

Klinsky,

Wetten,

Zanni-Ruiz

et al. 2023

Front. Cell Dev. Biol.

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Introduction: Cortical reaction is a secretory process that occurs after a spermatozoon fuses with the oocyte, avoiding the fusion of additional sperm. During this exocytic event, the cortical granule membrane fuses with the oocyte plasma membrane. We have identified several molecular components involved in this process and confirmed that SNARE proteins regulate membrane fusion during cortical reaction in mouse oocytes. In those studies, we microinjected different nonpermeable reagents to demonstrate the participation of a specific protein in the cortical reaction. However, the microinjection technique has several limitations. In this work, we aimed to assess the potential of cell-penetrating peptides (CPP) as biotechnological tools for delivering molecules into oocytes, and to evaluate the functionality of the permeable tetanus toxin (bound to CPP sequence) during cortical reaction.Methods: Arginine-rich cell-penetrating peptides have demonstrated the optimal internalization of small molecules in mammalian cells. Two arginine-rich CPP were used in the present study. One, labeled with 5-carboxyfluorescein, to characterize the factors that can modulate its internalization, and the other, the permeable light chain of tetanus toxin, that cleaves the SNAREs VAMP1 and VAMP3 expressed in mouse oocytes.Results: Results showed that fluorescent CPP was internalized into the oocyte cytoplasm and that internalization was dependent on the concentration, time, temperature, and maturation stage of the oocyte. Using our functional assay to study cortical reaction, the light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibited cortical granules exocytosis.Discussion: Results obtained from the use of permeable peptides demonstrate that this CPP is a promising biotechnological tool to study functional macromolecules in mouse oocytes.

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibits cortical reaction in mouse oocytes

Klinsky,

Wetten,

Zanni-Ruiz

et al. 2023

Front. Cell Dev. Biol.

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Dithiothreitol prevents the spontaneous release of cortical granules in in vitro aged mouse oocytes by protecting regulatory proteins of cortical granules exocytosis and thickening the cortical actin cytoskeleton

Wetten,

Klinsky,

Michaut

2024

Theriogenology

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In Celebration of Women in Molecular and Cellular Reproduction

2024

Frontiers Research Topics

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VAMPs sensitive to tetanus toxin are required for cortical granule exocytosis in mouse oocytes

Cited by 4 publications

References 57 publications

The light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibits cortical reaction in mouse oocytes

The light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibits cortical reaction in mouse oocytes

Dithiothreitol prevents the spontaneous release of cortical granules in in vitro aged mouse oocytes by protecting regulatory proteins of cortical granules exocytosis and thickening the cortical actin cytoskeleton

In Celebration of Women in Molecular and Cellular Reproduction

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