Theodor Boveri and the Formation of the Germ Line

Theodor Boveri (1862–1915) was the first person to observe an organism’s chromosomes throughout its development. In so doing, he discovered a fascinating feature in the development of the roundworm Parascaris aequorum (formerly known as Ascaris megalocephala). This nematode worm has only two chromosomes per haploid cell, allowing detailed observations of its individual chromosomes. The cleavage plane of the first embryonic division is unusual in that it is equatorial, separating the animal half from the vegetal half of the zygote (Figure 1). More bizarre, however, is the behavior of the chromosomes in the subsequent division of these first two blastomeres. The chromosomes in the animal blastomere fragment into dozens of pieces just before this cell divides. This phenomenon is called chromosome diminution, because only a portion of the original chromosome survives. Numerous genes are lost when the chromosomes fragment, and these genes are not included in the newly formed nuclei (Tobler et al. 1972; Müller et al. 1996).

Figure 1

Figure 1   Distribution of germ plasm during cleavage ofnormal and centrifuged zygotes of Parascaris. (A) In normal cleavage,the germ plasm is localized in the vegetalmost blastomere,as shown by the lack of chromosomal diminution in that particularcell. Thus, at the 4-cell stage, the embryo has a single stem cellfor its gametes. (B) When centrifugation is used to displace thefirst cleavage by 90°, both of the resulting cells have vegetal germplasm, and neither cell undergoes chromosome diminution. Afterthe second cleavage, both of these two cells give rise to germinalstem cells. (After Waddington 1966.)

Meanwhile, in the vegetal blastomere, the chromosomes remain normal. During second cleavage, the animal cell splits meridionally while the vegetal cell again divides equatorially. Both vegetally derived cells have normal chromosomes. However, the chromosomes of the more animally located of these two vegetal blastomeres fragment before the third cleavage. Thus, at the 4-cell stage, only one cell—the most vegetal—contains a full set of genes. At successive cleavages, nuclei with diminished chromosomes are given off from this vegetalmost line until the 16-cell stage, when there are only two cells with undiminished chromosomes. One of these two blastomeres gives rise to the germ cells; the other eventually undergoes chromosome diminution and forms more somatic cells. The chromosomes are kept intact only in those cells destined to form the germ line. If this were not the case, the genetic information would degenerate from one generation to the next. The cells that have undergone chromosome diminution generate the somatic cells.

Boveri has been called the last of the great observers of embryology and the first of the great experimenters. Not content with observing the retention of the full chromosome complement by the germ cell precursors, he set out to test whether a specific region of cytoplasm protects the nuclei within it from diminution. If so, any nucleus happening to reside in this region should remain undiminished. In 1910, Boveri tested this hypothesis by centrifuging Parascaris eggs shortly before their first cleavage. This treatment shifted the orientation of the mitotic spindle. When the spindle forms perpendicular to its normal orientation, both resulting blastomeres contain some of the vegetal cytoplasm (Figure 1B). Boveri found that after the first division, neither nucleus underwent chromosomal diminution. However, the next division was equatorial along the animal-vegetal axis. Here the resulting animal blastomeres both underwent diminution, whereas the two vegetal cells did not. Boveri concluded that the vegetal cytoplasm contains a factor (or factors) that protects nuclei from chromosomal diminution and determines germ cells.