Changing expression of chloride channels during preimplantation mouse development

Abstract

Plasma membrane chloride channels (ClCs) play important roles in a broad range of cellular processes including cell volume regulation, proliferation, and transepithelial transport, all of which are critical during preimplantation embryonic development. In this study, the molecular and functional expression of voltage-gated ClCs was analyzed throughout preimplantation development of the mouse conceptus. mRNA transcripts for all Clcn genes were detected. Only Clcn1 mRNA showed differential expression in the blastocyst, being detected in the trophectoderm but not in the inner cell mass. CLCN3 protein was detected at low levels in the cytoplasm and plasma membrane in 4-cell embryos and was localized to the apical plasma membrane of the trophoblasts in the blastocyst. Whole-cell patchclamp recordings demonstrated the presence of a DIDS-sensitive, outwardly rectifying Cl K current throughout development, with this conductance being large at the 1-cell, morula and blastocyst stages. A second DIDS-insensitive Cl K current, which was inactivated by membrane depolarization, was present in cells differentiating into the trophoblast lineage and during blastocyst expansion. Inhibition of the DIDS-sensitive current and the DIDS-insensitive current, with 9-AC, prevented blastocyst expansion.

Involvement of the TREK‐1 channel in human alveolar cell membrane potential and its regulation by inhibitors of the chloride current

“…To characterize the overall membrane current response, the following solutions were used: 9-acetylanthracene (9-AC; Tocris) was dissolved in ethanol and added to the perfusion solution to achieve a final concentration of 1 mM (Arnaiz, Johnson, Cook, & Day, 2013;Cronier, Bois, Hervé, & Malassiné, 1995).…”

“…4-(2-butyl-6,7-dichloro-2-cyclo pentylindan-1-on-5-yl) oxobutyric acid (DCPIB, Tocris, Bristol, UK) was dissolved in ethanol and added to the perfusion solution to achieve a final concentration of 10 μM (Sato-Numata, Numata, Inoue, & Okada, 2016). 9-acetylanthracene (9-AC; Tocris) was dissolved in ethanol and added to the perfusion solution to achieve a final concentration of 1 mM (Arnaiz, Johnson, Cook, & Day, 2013;Cronier, Bois, Hervé, & Malassiné, 1995). Barium chloride (BaCl 2 ; Merk, Darmstadt, DE) was dissolved in the perfusion solution to achieve a final concentration of 4 mM (Ma et al, 2011).…”

Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/B0AT1 Is Required for Optimal Preimplantation Embryo Development in Mice

“…Thus, in this study, Pro was taken up in B 0 AT1 knockout oocytes and embryos via a non-saturable route. This may be due to transport via channels that also conduct AAs, such as the volume-sensitive anion channel known to be present in oocytes [34][35][36].…”

Ion currents in embryo development

“…In particular, whole-cell patch-clamp recordings demonstrated the presence of a DIDS-sensitive, outwardly rectifying Cl -current throughout embryo development, with a large conductance at the zygote, morula, and blastocyst stages. A second DIDS-insensitive Cl -current was detected in cells differentiating into the trophoblast lineage and during blastocyst expansion (Arnaiz et al, 2013).…”