The development of lampbrush chromosome-type transcription in the early diplotene oocytes of Xenopus laevis: an electron-microscope analysis.

Oocytes of Xenopus laevis in pachytene and early diplotene of meiosis have been studied using the Miller spreading technique. Transcription first appears in germinal vesicles 25-40 micrometers in diameter, when the oocyte is in early diplotene. Transcription at this stage consists of arrays of short RNP transcripts, irregularly distributed along the DNP axis. Linear regression analysis has shown that many of these arrays are transcription units (Type I) with the transcripts having a common point of origin. The lengths of these early transcription units (mean = 7.06 +/- 5.06 micrometers), calculated from the linear regression data, are comparable to the lengths of transcription units from later stages, including Dumont stage 3. The polymerase granules of Type I transcription units are widely and irregularly spaced, having a mean spacing of 505 +/- 748 nm. More advanced transcription units (Type II, mean length = 8.72 +/- 3.77 micrometer) are usually found in the same chromosome set as the Type I units. Type II transcription units have closer and more regularly spaced polymerase granules than Type I transcription units (mean spacing = 92 +/- 49 nm). Both Type I and II transcription units have comparatively short RNP transcripts, the mean values for the slopes of their regression lines being 0.1336 and 0.1440 respectively. By the time the germinal vesicles are about 50-60 micrometers in diameter the transcription units have a quite different morphology (Type III). The lengths of the Type III transcription units are comparable to the Type I and II units, the mean length being 6.34 +/- 4.03 micrometers. The spacing of the polymerase granules in the Type III units is closer and more regular than the earlier stages (70 +/- 40 nm). Another significant difference between Type III and Types I and II transcription units is a decrease in the foreshortening of the Type III RNP transcripts. The mean slope of the regression lines for Type III transcription units is 0.2439. The morphological appearance of the Type III transcription unit is virtually identical to that of the transcription units from Dumont stage 3 oocytes, both with respect to the length and the spacing of the polymerase granules. However, the transcripts in Type III transcription units are still more foreshortened than those of Dumont stage 3 oocytes, having mean regression slopes of 0.4728. From the data obtained in the present study, it has been concluded that the pattern of lampbrush-type transcription is virtually fully established by the time most germinal vesicles are about 50 micrometers in diameter.