Blattella germanica.
Unlike the spermatogonia of Stenopelmatus, those of Blattella have both a faintly-staining nucleolus and a deeply-staining chromatin element (x), and moreover the two are always closely associated (figs. 95, 96). The number of chromatin elements in the equatorial plate of late spermatogonial mitoses is 23 (fig. 97). Later events indicate that one of the 23 is the element x, but it is impossible to distinguish it here. Figure 98 is a very early stage of the spermatocyte of the first order, showing the element x as a U-shaped body. The centrosome was also conspicuous in all of the cells of this group. The spireme here, as also in figure 99, is fine and closely interwound. In figure 99 and again in figure 100 the element x is joined to the spireme as it is throughout the spireme stage. In the "bouquet" or "polarized" stage the combined nucleolus and element x are always at one side of the group of loops and down very close to the base of the figure (figs. 101, 103). In figure 102 most of the loops are cut across. The stage shown in figures 104 and 105 is a later one than that just described. Here we have again a continuous spireme connected with the element x, making it seem improbable that the bivalent chromosomes are really separated in the bouquet stage. Figure 106 gives some of the variations in form of the combined nucleolus and element x. The last of the five figures was taken from a giant cell containing at least twice the usual amount of chromatin. In one giant cell four unusually large combinations of this kind were found, and a corresponding amount of chromatin in the spireme. In figure 107 one sees the spireme divided into segments still joined by linin bridges. In figure 108 similar segments may be seen, one of them showing a longitudinal split. The element x is present, but the nucleolus has disappeared. In many cases the split, if it appears at all, closes quickly and the chromosome bends in U-shape, as in figure 109, plate IV. This figure also shows two centrosomes (c). In other cases the split persists as in figure 110 and leads to the formation of crosses of a tetrad character (figs. 111, 112, 113), as in Stenopelmatus and many other insects. Figures 114 to 117 show later stages of the U-shaped chromosomes. Perfect rings are rare. All sorts of variations are seen, broad and narrow U-shapes, rings split at one point or the opposite points, a U split at the bottom (fig. 114), pairs of parallel rods (fig. 115), and occasionally rods constricted in the middle and showing a longitudinal split in each half, as in figure 116. Figure 117 shows different views of the split rings. Apparently all of these forms straighten out so that the two components of the bivalent chromosome stand end to end as dumbbells or compressed crosses in the metaphase of the first maturation spindle (figs. 123-125). The element x remains concentrated and more or less spherical in form. Figures 118-122 are equatorial plates, with x absent in figure 120, in the same plane as the 11 other chromosomes in figure 119, far to one side in figure 118, and near one pole of the forming spindle in figure 122. It is also shown in various positions with regard to the spindle in figures 123 to 126 and 128 to 132. In figure 125 it is apparently double, and again in figure 129. In figure 130 one lagging chromosome shows the dyad nature of the products of the division of the tetrad. In this form there can be no doubt that reduction occurs in the first spermatocyte division. The element x is very often concealed by the polar aggregation of chromatin, but it is sometimes as conspicuous as in figures 131 and 132. The spermatocytes of the second order go into a complete resting stage before they are completely separated, and one of a pair shows the element x, while it is lacking in the other (fig. 133). At the close of the resting stage the chromosomes appear as 11 pairs of rods of considerable length, which gradually shorten and thicken and usually bend at the center, forming U's or V's (figs. 134-138). In one stage these double U's look much like tetrads (fig. 138). The rods straighten again as they shorten still more (fig. 139), become more closely approximated, and finally form dumbbells, as in figure 141.
The element x is, of course, present in only one-half of these nuclei. In the equatorial plate, figure 142, it is absent; in figure 143 it is present, but can not be distinguished from the other chromosomes, while in figure 144 it is rendered conspicuous by its spherical form and isolated position. In only a few cases has it been possible to distinguish x in the spindle. Figures 146 and 147 show two of these cases where this element is clearly double and of different form from the other chromosomes. It is probable that it divides and so goes into one-half of all of the spermatids, as in McClung's typical cases of the accessory chromosome. Figure 145 shows the usual appearance of the other chromosomes in metaphase. The two spermatids of a pair are always alike so far as any evidence of the presence of the element x is concerned (fig. 148). Figure 149 is an exceptional case, where one chromatin element (possibly x) has evidently divided late and been left out in the cytoplasm; a smaller chromatin granule is also present in the cytoplasm of each spermatid. All of the spermatids, as in Stenopelmatus, develop a deeply-staining body, which, however, in this case is usually centrally located and often appears double (figs. 150-152).
The spindle-remains (Spindelreste) forms a very conspicuous body at one side of the nucleus in the spermatids, and occasionally a mass of chromatin, probably due to imperfect mitosis, is found near the spindle-substance (fig. 150). The mass of spindle-substance at first appears structureless, but soon assumes the condition shown in figures 150 to 152. In one individual many of the spermatids had two balls of spindle-material (fig. 152), and the resulting later stages were double-tailed (fig. 153). Figure 156 shows how the spindle-substance goes into the tail and gradually disappears as the tail lengthens.
The centrosome is evidently applied to the nuclear membrane, as in Stenopelmatus, and the middle-piece is developed in connection with it, as in figures 156-157, 154-155, 158-160. The element x of the spermatids gradually disappears (figs. 150, 159). An acrosome develops at the anterior end, the head condenses and lengthens, and we have the ripe spermatozoön (fig. 161). The tail is very long and is shown only in part.
Of the forms studied, Blattella alone has many degenerate spermatozoa. Some follicles have none, others a number varying perhaps from one-fourth to three-fourths of the whole number. No evidence of degeneracy was detected among the young spermatids up to the stage shown in figures 154-155, where a few like figure 162 were found. Most of the degenerate forms occur among the nearly ripe spermatozoa or in the sperm-ducts. Such are shown in figures 163 to 168. The chromatin is strangely broken up into irregular clumps, and probably no two of these degenerate sperm-heads can be found which are alike. The tails are always imperfect. The distribution and varying numbers of these degenerate spermatozoa make it impossible to interpret their condition as due to the absence of the accessory chromosome, as Miss Wallace does in the spider. The only probable explanation, it seems to me, is imperfect mitosis. Cases where more or less chromatin is left behind in the cytoplasm, especially in the first spermatocyte mitosis, are very common, and such cases as those shown in figures 149 and 150 are not rare. The giant cells, so far as I have been able to trace them, do not develop into spermatozoa.
The most important points are:
(1) The presence of the element x in the spermatogonia, closely associated with the nucleolus.
(2) The uneven number of chromatin elements in the metaphase of spermatogonial divisions.
(3) The connection of the element x with the spireme up to the stage where the spireme segments to form the bivalent chromosomes.
(4) The varied character of the tetrads, showing the first spermatocyte division to be a reducing division in the sense that it separates whole chromosomes.
(5) The fact that the element x fails to divide in the first maturation division, does divide in the second, but can not be traced beyond the equatorial plate of the latter mitosis.
(6) The similarity of all the normal spermatids, though one-half of them must contain the element x, the other half not.
(7) The varying and often large number of degenerate spermatozoa.
An attempt was made to determine the somatic number of chromosomes. The dividing cells of the follicles of young eggs seemed to afford the most favorable material, but even here there was so much overlapping of the ends of the chromosomes that it was impossible to be absolutely certain of the number. In the two most favorable cases 23 were counted (fig. 94). This differs from McClung's count for similar cases among the Orthoptera, and Sutton's for Brachystola magna. The eggs have so far resisted all efforts to learn what part the odd chromosome may play in fertilization.