Data on the concentrations of etoposide, PSC833, BAPTA-AM, and cycloheximide that do not compromise the vitality of mature mouse oocytes, parthenogenetically activated and fertilized embryos

These data document the vitality of mature mouse oocytes (Metaphase II (MII)) and early stage embryos (zygotes) following exposure to the genotoxic chemotherapeutic agent, etoposide, in combination with PSC833, a selective inhibitor of permeability glycoprotein. They also illustrate the vitality of parthenogenetically activated and fertilized embryos following incubation with the calcium chelator BAPTA-AM (1,2-Bis(2-aminophenoxy)ethane- N,N,N′,N′-tetraacetic acid tetrakis (acetoxymethyl ester)), cycloheximide (an antibiotic that is capable of inhibiting protein synthesis), and hydrogen peroxide (a potent reactive oxygen species). Finally, they present evidence that permeability glycoprotein is not represented in the proteome of mouse spermatozoa. Our interpretation and discussion of these data feature in the article “Identification of a key role for permeability glycoprotein in enhancing the cellular defense mechanisms of fertilized oocytes” (Martin et al., in press) [1].


Specifications
Mouse oocytes and zygotes were treated with etoposide (100 mg/ml) for 15 min when appropriate. Those used in the examination of permeability glycoprotein (PGP) efflux activity were pretreated with either the PGP inhibitor PSC833 for 15 min (5 mM) or with BAPTA-AM (5 mM)), or cycloheximide (20 mg/ml) for 4 h during the allotted activation/fertilization period.

Experimental features
Mouse oocytes and spermatozoa were harvested and zygotes or parthenotes produced via IVF or strontium chloride chemical activation, respectively. Oocytes were treated with etoposide and PSC833 in combination (a selective inhibitor of PGP), BAPTA-AM or cycloheximide. The cytotoxicity of these drugs was evaluated by labeling of the cells with a standard vitality reagent for 15 min at 37°C.

Data accessibility
All relevant data are presented within this article

Value of the data
These data provide valuable insight into the maintenance of mature mouse oocyte and zygote vitality following genotoxic insult with etoposide (100 mg/ml); a chemotherapeutic agent that elicits a potent inhibition of topoisomerase II α action. Similarly, these data indicate that selective pharmacological inhibition of permeability glycoprotein (PGP) with PSC833 (5 mM), as well as incubation of oocytes in BAPTA-AM (5 mM) and cycloheximide (20 mg/ml) for periods of up to 4 h following insemination with spermatozoa or activation with strontium, does not adversely affect oocyte or embryo vitality.
This information is of use to the scientific community as it establishes concentrations of various pharmacological reagents that can be utilized without compromising oocyte and embryo viability.
Finally, these data provide evidence that mouse spermatozoa do not harbor PGP within their proteome, thus discounting the possibility of a paternal contribution to elevated levels of PGP found in the zygote.

Data
The files included in this article comprise vitality profiles of mouse MII stage oocytes, chemically activated and fertilized zygotes following exposure to etoposide (100 mg/ml) in combination with PSC833 (5 mM) (Fig. 1), cycloheximide (20 mg/ml), BAPTA-AM (5 mM) or hydrogen peroxide (1 mM) (Fig. 3). This latter treatment was included as a positive control. Immunoblots of mouse sperm lysates with of anti-PGP antibodies are also included in this article (Fig. 2).

Reagents
Reagents were purchased from Sigma Aldrich (St Louis, MO, USA) unless otherwise stated. Antipermeability glycoprotein (PGP; ab170904) antibody used for immunoblotting was procured from Abcam (Cambridge, England, UK).

Strontium activation and in vitro fertilization (IVF)
Parthenogenic activation was stimulated by incubation of cumulus free oocytes in calcium free KSOM medium supplemented with 10 mM strontium chloride for 4 h. In vitro fertilization was carried out in human tubal fluid (HTF) medium containing 1 mM reduced glutathione (GSH). Recovered oocytes were co-incubated with 2 Â 10 5 capacitated spermatozoa for 4 h at 37°C and successful fertilization (or activation in the case of parthenotes) was assessed by recording the extrusion of the second polar body and/or pronucleus formation [4].

Assessment of factors influencing permeability glycoprotein (PGP) expression
A number of drugs were used to investigate the factors responsible for the notable increase in PGP labeling following fertilization and activation [1]. The relative contribution of protein translation was analyzed by the inclusion of cycloheximide (20 mg/ml), an antibiotic with the ability to block protein synthesis. Alternatively, the role of activation-associated cytosolic calcium oscillations was assessed in the presence of the calcium chelator, BAPTA-AM (5 mM).

Etoposide treatment and cellular vitality
MII oocytes, parthenotes and pronuclear stage zygotes, were exposed to etoposide (100 mg/ml for 15 min at 37°C), a topoisomerase II α inhibitor capable of eliciting DSB DNA damage. The cytotoxicity Fig. 1. Co-incubation with etoposide and valspodar (PSC833), a selective inhibitor of PGP efflux activity did not see a loss in oocyte vitality. The cytotoxicity of etoposide and PSC833 were evaluated using as a standard far red detectable live/dead vitality reagent. Analysis confirmed that in no instance was there an associated decrease in oocyte vitality following treatment exposure n ¼3.
of etoposide, PSC833 [5,6], BAPTA-AM and cycloheximide at the specific concentrations employed in this investigation were evaluated by labeling of the cells with a far red detectable live/dead vitality reagent (Thermo Fisher Scientific, Waltham, MA, USA).

Statistical analysis
Image processing was achieved using the public sector program, Image J (National Institute of Health, Bethesda, MD, USA). Statistical significance was determined using JMP software (version 10.0.0, SAS Institute, NC, USA). Each experiment was conducted on a minimum of three biological replicates and expressed as the mean 7s.e.m. Differences with a value of Po 0.05 were considered statistically significant.

Transparency document. Supplementary material
Transparency data associated with this article can be found in the online version at http://dx.doi. org/10.1016/j.dib.2016.07.046.