Effects of 6-dimethylaminopurine on microtubules and putative intermediate filaments in sea urchin embryos

Dufresne, Louise; Néant, Isabelle; St‐Pierre, Johanne; Dubé, François; Guerrier, P.

doi:10.1242/jcs.99.4.721

Cited by 22 publications

(1 citation statement)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The three inducers utilized in this experiment were CB, 6-DMAP and salinity, which were widely used in inducing triploid and tetraploid shell shes (Gérard et The effect on fertilized eggs can be virtually instantaneous, thus the window for treating fertilized eggs required to be strictly controlled (Peachey and Allen 2016). 6-DMAP acts as an inhibitor of protein phosphorylation and protein kinase activity (Dufresne et al 1991). Many experts have been able to induce shell sh polyploidization using 6-DMAP with an e ciency comparable to that of CB (Peachey and Allen 2016; Gérard et al 1999).…”

Section: The Tetraploid Rate Was Signi Cantly Affected With the Induc...mentioning

confidence: 99%

Comparative study on tetraploid induction of the Fujian oyster Crassostrea angulata utilizing three typical methods

Liang

Geng

Bai

et al. 2023

Preprint

View full text Add to dashboard Cite

The sterility advantages of triploids make them paramount for oyster aquaculture. Current hatcheries for triploid industrial breeding utilize tetraploids as male parents, and thus tetraploids are a core asset for triploid oyster culture. The oyster Crassostrea angulata is the most productive oyster species in China, however, the optimal conditions for tetraploid induction have never been examined. In this study, we investigated the effectiveness of three induction methods (Method Ⅰ, 2n ♀ × 2n ♂ + inducer abbreviated as DD + ind; Method Ⅱ, 3n ♀ × 2n ♂ + inducer abbreviated as TrD + ind; Method Ⅲ, 2n ♀ × 4n ♂ + inducer abbreviated as DT + ind) and three inducers (salinity; cytochalasin B, CB; and 6-dimethylaminopurine, 6-DMAP) for tetraploid induction in C. angulata. The optimal induction conditions for the three induction methods were a CB concentration of 0.5 mg/L treated for 20 min, 0.75 mg/L CB for a constant induction treatment of 20 min and 6-DMAP at a concentration of 100 mg/L for 20 min, respectively. When the optimal induction conditions were applied to a single pair of oysters, the D-larval tetraploid rate for the three methods were 18.93%, 58.32% and 51.61%, respectively. The survival rates of the experimental groups were only 10.36%, 28.07% and 21.99%, respectively, at day 30. The tetraploid rates for Method Ⅰ and Method Ⅲ, although decreased significantly over time, still reached 11.81% and 21.99% at 30 days, and 8% and 16% at 180 days of age, respectively. Tetraploid rates for Method Ⅱ were 58.32% − 71.28% at the larval stage and 80% at day 180. In summary, in spite of very different induction rates, viable tetraploid C. angulata can be harvested in all three methods. Method Ⅱ (TrD + inducer) was regarded as the optimal method to induce C. angulata tetraploids, and the optimal induction condition was a CB concentration of 0.75 mg/L and treated for 20 min under this method.

show abstract

Section: The Tetraploid Rate Was Signi Cantly Affected With the Induc...mentioning

confidence: 99%

Comparative study on tetraploid induction of the Fujian oyster Crassostrea angulata utilizing three typical methods

Liang

Geng

Bai

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

Effects of cell-cycle-arrest agents on cleavage and development of mouse embryos

Samaké

Smith

1996

J. Exp. Zool.

View full text Add to dashboard Cite

In mammals, there are no reliable methods for synchronizing cell division of early embryos without reducing their ability to develop into blastocysts and fetuses. The present study was undertaken to examine the in vitro inhibition of cell division of four‐cell mouse embryos by cell cycle arrest agents. The reversibility of the agents was also tested by examining the developmental ability of treated embryos. Four‐cell mouse embryos obtained at 54 hr post‐human chorionic gonadotrophin (post‐hCG) were cultured for 4, 8, 12, or 16 hr in media supplemented with either nocodazole, an inhibitor of tubulin polymerization, 6‐dimethylaminopurine (6‐DMAP), an inhibitor of maturation promoting factor (MPF) activation, or aphidicolin, a specific inhibitor of DNA polymerase alpha. Reversibility and toxicity of these agents were both dose and time dependent. For all three agents, prolonging cleavage arrest for 8 or 16 hr (at the effective concentrations) caused embryo lethality. Although nocodazole treatment was least cytotoxic, 6‐DMAP and aphidicolin concentrations which induce cleavage arrest were detrimental to development beyond the blastocyst stage. The results of this study show that the development of embryos treated with these three cell‐cycle‐arrest agents is dose and incubation time dependent. Toxic effects beyond the blastocyst stage could only be minimized for nocodazole by reducing the exposure time of treatment and concentration of the mitotic inhibitor. However, these results render doubt on the usefulness of 6‐DMAP and aphidicolin for synchronization studies leading to embryo transfer procedures. © 1996 Wiley‐Liss, Inc.

show abstract

Phosphorylation protects sperm-specific histones H1 and H2B from proteolysis after fertilization

Morín¹,

Acuña²,

Díaz³

et al. 2000

J. Cell. Biochem.

View full text Add to dashboard Cite

At intermediate stages of male pronucleus formation, sperm-derived chromatin is composed of hybrid nucleoprotein particles formed by sperm H1 (SpH1), dimers of sperm H2A-H2B (SpH2A-SpH2B), and a subset of maternal cleavage stage (CS) histone variants. At this stage in vivo, the CS histone variants are poly(ADP-ribosylated), while SpH2B and SpH1 are phosphorylated. We have postulated previously that the final steps of sperm chromatin remodeling involve a cysteine-protease (SpH-protease) that degrades sperm histones in a specific manner, leaving the maternal CS histone variants unaffected. More recently we have reported that the protection of CS histones from degradation is determined by the poly(ADP-ribose) moiety of these proteins. Because of the selectivity displayed by the SpH-protease, the coexistence of a subset of SpH together with CS histone variants at intermediate stages of male pronucleus remodeling remains intriguing. Consequently, we have investigated the phosphorylation state of SpH1 and SpH2B in relation to the possible protection of these proteins from proteolytic degradation. Histones H1 and H2B were purified from sperm, phosphorylated in vitro using the recombinant alpha-subunit of casein kinase 2, and then used as substrates in the standard assay of the SpH-protease. The phosphorylated forms of SpH1 and SpH2B were found to remain unaltered, while the nonphosphorylated forms were degraded. On the basis of this result, we postulate a novel role for the phosphorylation of SpH1 and SpH2B that occurs in vivo after fertilization, namely to protect these histones against degradation at intermediate stages of male chromatin remodeling.

show abstract

Effects of 6-dimethylaminopurine on microtubules and putative intermediate filaments in sea urchin embryos

Cited by 22 publications

References 43 publications

Comparative study on tetraploid induction of the Fujian oyster Crassostrea angulata utilizing three typical methods

Comparative study on tetraploid induction of the Fujian oyster Crassostrea angulata utilizing three typical methods

Effects of cell-cycle-arrest agents on cleavage and development of mouse embryos

Phosphorylation protects sperm-specific histones H1 and H2B from proteolysis after fertilization

Contact Info

Product

Resources

About