Van Winkle L J, Patel M, Wasserlauf H G, Dickinson H R, Campione A L
Department of Biochemistry, Midwestern University, Chicago College of Osteopathic Medicine, Downers Grove, IL 60515.
Biochim Biophys Acta. 1994 May 11;1191(2):244-55. doi: 10.1016/0005-2736(94)90175-9.
Taurine was shown recently to increase the frequency at which 2-cell mouse conceptuses develop into blastocysts in vitro. For this reason and because taurine helps cells adapt to external stresses, we studied transport of this and related amino acids by preimplantation mouse conceptuses. The most conspicuous component of taurine transport in conceptuses at the 1-cell through blastocyst stages of development was both Na(+)- and Cl(-)-dependent. This Na(+)- and Cl(-)-dependent transport system interacted relatively strongly with beta- but not alpha-amino acids. By these criteria, transport system beta is responsible for Na(+)-dependent taurine transport in preimplantation mouse conceptuses. Moreover, detection of mRNA encoding the taurine transport protein (TAUT) in early conceptuses supports the theory that TAUT is a major component of system beta. Transport of taurine by system beta in 1-cell conceptuses was slower in hypotonic than in hypertonic media, whereas the reverse was true for system beta in blastocysts. In contrast, hypotonically stimulated Na(+)-independent taurine transport was, of course, more rapid in hypotonic than in hypertonic media in both 1-cell conceptuses and blastocysts. Transport via this hypotonically stimulated process also showed no sign of saturation by up to 10 mM taurine. Hypotonically stimulated taurine transport appeared transiently in 1-cell conceptuses under hypotonic conditions until they had recovered their initial volumes. Hence, we suggest that a decrease in taurine uptake via system beta and an increase in taurine exodus via the Na(+)-independent, nonsaturable transport process could contribute to the regulatory volume decrease in 1-cell conceptuses in hypotonic medium. Since taurine uptake by system beta in blastocysts is, however, higher in hypotonic than in hypertonic media, taurine uptake by system beta in blastocysts might intensify a tendency to increase cell volume in hypotonic medium. Such an increase in taurine uptake could further favor anabolic changes associated with cell swelling. In addition to contributing to regulation of cellular volume and perhaps metabolism, the hypotonically stimulated Na(+)-independent transport processes in early embryos have novel characteristics. Hypotonically stimulated Na(+)-independent taurine transport was inhibited by niflumate, N-ethylmaleimide and NaN3 but not by furosemide, iodoacetate, KCN, ouabain or alpha- or beta-amino acids. Furthermore, 4,4'-diisothiocyanostilbene-2,2'-disulfonate inhibited this transport in 1-cell conceptuses but not in blastocysts. Hence, different hypotonically stimulated Na(+)-independent taurine transport processes appear to be present in 1-cell conceptuses vs. blastocysts. The functions of these and other instances of developmental regulation of expression of transport processes in preimplantation conceptuses remain largely to be elucidated.(ABSTRACT TRUNCATED AT 400 WORDS)
最近研究表明,牛磺酸能提高体外培养的2细胞期小鼠胚胎发育成囊胚的频率。鉴于此,且由于牛磺酸有助于细胞适应外界压力,我们研究了植入前小鼠胚胎对这种氨基酸及相关氨基酸的转运情况。在从1细胞期到囊胚期的胚胎发育过程中,牛磺酸转运最显著的成分是依赖Na⁺和Cl⁻的。这种依赖Na⁺和Cl⁻的转运系统与β氨基酸相互作用较强,而与α氨基酸相互作用较弱。根据这些标准,转运系统β负责植入前小鼠胚胎中依赖Na⁺的牛磺酸转运。此外,在早期胚胎中检测到编码牛磺酸转运蛋白(TAUT)的mRNA,这支持了TAUT是系统β主要成分的理论。系统β在1细胞期胚胎中对牛磺酸的转运在低渗培养基中比在高渗培养基中慢,而在囊胚中则相反。相比之下,低渗刺激的不依赖Na⁺的牛磺酸转运,在1细胞期胚胎和囊胚中,低渗培养基中当然比高渗培养基中更快。通过这种低渗刺激过程的转运,在牛磺酸浓度高达10 mM时也没有饱和迹象。低渗刺激的牛磺酸转运在低渗条件下短暂出现在1细胞期胚胎中,直到它们恢复初始体积。因此,我们认为通过系统β的牛磺酸摄取减少以及通过不依赖Na⁺的、不可饱和的转运过程的牛磺酸外流增加,可能导致低渗培养基中1细胞期胚胎的调节性体积减小。然而,由于系统β在囊胚中对牛磺酸的摄取在低渗培养基中高于高渗培养基,系统β在囊胚中对牛磺酸的摄取可能会增强低渗培养基中细胞体积增加的趋势。这种牛磺酸摄取的增加可能进一步有利于与细胞肿胀相关的合成代谢变化。除了有助于调节细胞体积以及可能的代谢外,早期胚胎中低渗刺激的不依赖Na⁺的转运过程具有新的特征。低渗刺激下不依赖Na⁺的牛磺酸转运受到尼氟灭酸、N - 乙基马来酰胺和NaN₃的抑制,但不受速尿、碘乙酸、KCN、哇巴因或α或β氨基酸的抑制。此外,4,4'-二异硫氰基芪-2,2'-二磺酸抑制1细胞期胚胎中的这种转运,但不抑制囊胚中的转运。因此,1细胞期胚胎和囊胚中似乎存在不同的低渗刺激下不依赖Na⁺的牛磺酸转运过程。植入前胚胎中这些以及其他转运过程表达的发育调控实例的功能在很大程度上仍有待阐明。(摘要截断于400字)