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A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Published on: February 6, 2018
Fyn kinase-tubulin interaction during meiosis of rat eggs
A Talmor-Cohen1, R Tomashov-Matar, W B Tsai
1Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University, Ramat-Aviv 69978 Tel-Aviv, Israel.
Abstract:
Prior to fertilization, the spindle of vertebrate eggs must remain stable and well organized during the second meiotic meta-phase arrest (MII). In a previous study we have determined that the completion of meiosis is a Src family kinase (SFK)-dependent event. In the current study we have used the SFK inhibitors, SU6656 and PP2, and demonstrated that inhibition of SFKs caused the formation of a disorganized spindle. The observation that proper organization of an MII spindle is an SFK-dependent process, combined with our previous finding that Fyn kinase is localized at the microtubules (MTs), prompted us to examine the potential role of Fyn in MT signaling. Our results show an association between Fyn and tubulin, the ability of Fyn to phosphorylate tubulin in vitro and stimulation of meiosis completion by injection of a constitutively active form of Fyn (CAF). We suggested that SFKs mediate significant functions during the organization of the MII spindle. In view of CAF injection experiments, and of the pronounced concentration of Fyn kinase at the spindle, we propose that Fyn may play an important role in some aspects of the spindle functions, possibly those involving the MTs.
Insights
Src family kinases (SFKs) are crucial for maintaining vertebrate egg spindle stability during meiosis II arrest. Fyn kinase, an SFK, directly interacts with tubulin, suggesting a key role in microtubule organization for successful cell division.
Area of Science:
- Cell Biology
- Molecular Biology
- Developmental Biology
Background:
- Vertebrate egg spindle organization is essential for meiotic arrest at metaphase II (MII).
- Src family kinases (SFKs) regulate meiosis completion.
- Fyn kinase is localized to microtubules (MTs) within the spindle.
Purpose of the Study:
- To investigate the role of Fyn kinase in microtubule (MT) signaling and MII spindle organization.
- To determine if Fyn kinase directly influences tubulin and MTs.
Main Methods:
- Utilized SFK inhibitors (SU6656, PP2) to assess spindle organization.
- Examined Fyn kinase association with tubulin.
- Performed in vitro tubulin phosphorylation assays by Fyn.
- Injected constitutively active Fyn (CAF) to stimulate meiosis completion.
Main Results:
- SFK inhibition led to disorganized MII spindles.
- Demonstrated physical association between Fyn and tubulin.
- Showed Fyn phosphorylates tubulin in vitro.
- CAF injection stimulated meiosis completion.
Conclusions:
- SFKs are critical for MII spindle organization.
- Fyn kinase plays a significant role in spindle functions, likely through microtubule interactions.
- Fyn's phosphorylation of tubulin suggests a direct mechanism in MT regulation during meiosis.
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