Xenopus laevis oocyte G alpha subunits mRNAs. Detection and quantitation during oogenesis and early embryogenesis by competitive reverse PCR.
The expression of mRNAs coding for different Xenopus laevis oocyte G alpha subunits was analyzed by the PCR technique. Using the nucleotide sequences of five previously cloned cDNAs for oocyte G alpha subunits [FEBS Lett. 244, 188-192, 1989; FEBS Lett. 268, 27-31, 1990] and the highly sensitive reverse PCR reaction we found that G alpha o, G alpha i-1, G alpha i-3 and G alpha s species are present in oocyte stage VI, G alpha o mRNA being the most abundant transcript. G alpha o mRNA was further quantitated through oogenesis, unfertilized eggs and early embryogenesis stages by a competitive PCR reaction using an 'in vitro' deleted G alpha o mRNA as the internal standard. Using this approach we found that Xenopus G alpha o mRNA levels were constant during oogenesis and unfertilized eggs at a concentration of 3.5 pg of mRNA/stage (5 x 10(5) molecules) and diminish gradually during early embryogenesis, reaching a level of 0.3 pg in the gastrula stage. These findings show that oocyte G alpha o, and perhaps the rest of the alpha subunits, are expressed as maternal mRNAs and could play an important role in signal transduction at the beginning of oocyte cell differentiation.
Oñate
A
A
Departamento de Bioquímica, Universidad de Chile, Santiago.
Herrera
L
L
Antonelli
M
M
Birnbaumer
L
L
Olate
J
J
D000818:Animals; D001483:Base Sequence; D019204:GTP-Binding Proteins / Q000235:genetics*; D015870:Gene Expression; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D009865:Oocytes / Q000502:physiology; D009866:Oogenesis; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D014982:Xenopus laevis / Q000196:embryology* / Q000235:genetics
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D019204:GTP-Binding Proteins