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The cytoplasmic structures in the spermioteleosis of Bruchus quadrimaculatus were studied, using various fixatives and stains including vital and smear techniques. Mitochondria appear in all stages following spermatogonia, exhibiting characteristic appearances during mitoses and in the spermatids. The [central substance] in the halves of the nebenkern becomes localized along the edges of the outgrowing [ribbons] as the source of the marginal filament. The undulating membrane apparently is formed by the fusion of these [ribbons.] The axial filament arises from the distal centriole, being permanently attached thereto, and lies along one edge of the undulating membrane of the tail. Golgi bodies are first observed in the secondary spermatocyte; they behave characteristically during spermiogenesis, producing the acrosome and leaving a residuum, the Golgi remnant, to be sloughed off with the cytoplasm from the tail. Chromatoid bodies occur in some spermatids but not in others. They migrate backward along the axial filament to be rejected with the Golgi remnant.

The chondriosomes are first recognizable as two clusters in contact with the nuclear wall. Beneath each cluster lies a chromosome. This relationship is constant and indicates that these two chromosomes play a part in the growth and development of the chondriosomes. The two masses fuse and the single mass grows considerably during the early growth period. Later it breaks up into a number of threads which become rings in the late growth period. The rings fuse into the large nebenkern which plays the usual role in the formation of the tail. The Golgi material is first seen outside of the chondriosomal cap. Early in the growth period, it breaks up into Golgi bodies which remain distributed in the cytoplasm during the growth period and spermatocyte divisions. About the midgrowth period a large number of spheres suddenly arise. Since they later fuse to form the idiosome, they are called the proidiosomal spheres. These spheres, which may originate in the Golgi bodies, remain scattered in the cytoplasm during the growth period and spermatocyte divisions. In the spermatid the Golgi bodies collect about the idiosome to form the acroblast. The Golgi remnants pass into the cytoplasm of the tail, while the acrosome elongates into a tail-like structure at the anterior end. The centriole was followed with unbroken continuity from the midgrowth period into the middle-piece of the sperm.

Mitochondria in early spermatids of many insects aggregate and form a round body, the nebenkern. The nebenkern undergoes a structural differentiation and then divides into two separate equal-sized bodies. In the present study, nebenkerns of Murgantia histrionica, a Hemipteran insect, were reconstructed using electron micrographs of serial sections to determine how the mitochondria transform into the two separate bodies. Newly formed nebenkerns are made of one piece, an anastomosis of rod-like segments. Some segments interconnect to join networks of rings. Each network interlocks with another similar network, but networks which interlock are connected with each other by other segments of the nebenkern. Later, the entire nebenkern is made of two unconnected and interlocked networks of rings. The nebenkern appears to remain bipartite during subsequent differentiation. Since the two pieces are interlocked, breaks must occur before the pieces can separate. As breaks occur, each network transforms into a set of curved sheets, producing a nebenkern made of four concentric layers. The three outer layers are each made of two curved sheets which surround a bipartite central core. The surface sheets meet at a furrow in the surface of the nebenkern; segments in each layer are roughly symmetrical with each other about the plane in which the furrow lies. Rod-like segments join alternate segments. The number of layers then decreases to three, and later, to two. These nebenkerns resemble four-layered nebenkerns, but fewer connections between alternate segments are present. The two pieces constituting the nebenkern probably separate after most of the latter connections disappear. Hypotheses to account for the observed changes in nebenkern structure are presented.

The nuclear cap in the spermatogonial and early spermatocyte cells of Gelastocoris is an aggregate of closely packed mitochondria with their long axes perpendicular to the nuclear membrane. Eventually in the early growth period, the mitochondria move from the cap and appear to become more or less equally distributed in the cytoplasm where they remain until their fusion in the spermatid to form the nebenkern. The Golgi complex consists of clusters of lamellae and vesicles, the Golgi bodies. Granules form within the vesicles, increase in size, move from their place of origin and become distributed at random in the cytoplasm. They are the pro-acrosomal granules and at the end of the growth period fuse to form the proacrosome, about which Golgi bodies collect. The Golgi bodies, however, never fuse into an acroblast. At one end of the oval-shaped pro-acrosome is a small dark body and a less dense vesicle the future of which is uncertain. The dark body eventually occupies a position at the tip of the acrosome. The pro-acrosome, after moving to the side of the nucleus opposite the nebenkern, differentiates into the acrosome which elongates into a tail-like structure. The nuclear membrane of some spermatocytes may appear wave-like in cross section, with the crest and trough different in appearance. Near the membrane and in the troughs of the waves large clusters of granules are frequently present. Similar clusters may be found elsewhere in the cytoplasm. Presumably they had their origin near the membrane but this is not conclusive. Bodies of indeterminate origin and structure may be present in the cytoplasm. They could be lysosomes but evidence is lacking. In late spermatocytes and in spermatids, a group of ten or twelve granules is present. They are smaller than the pro-acrosomal granules, are always closely associated and pass as a group into the tail. Their significance is unknown. The endoplasmic reticulum is typical of cells in general. There are no granule accumulations within the vesicles as in some secretory cells. Vesicles of various shapes and sizes are present within the centrosphere of the first meiotic division. While their location is similar to that of the centriole, the identity of the vesicles is uncertain.

The axial filament of Sciara coprophila does not conform to the usual 9 + 2 filament pattern but consists, rather, of as many as 76 pairs of filaments which decrease in number from the anterior to the posterior region of the sperm. It is first seen at the base of the head in the shape of an indented oval. The axial filament varies in configuration along the remaining length of the sperm as one whorl or two connected whorls of filament pairs. The other structures of the sperm revealed by the light and electron microscopes are a homogeneous, dense, spear-shaped nucleus, a row of spherical dense bodies in the middle piece enclosed by the axial filament and of unknown nature and function and a single mitochondrial derivative. The mitochondrial nebenkern derivative consists of a large electron transparent region bordered by cristae and a smaller paracrystalline region located adjacent to the axial filament. The derivative arises as paracrystalline material in a medial nuclear indentation. The electron transparent material is first seen at the anterior end of the middle piece. Unlike other known insect sperm, but reminiscent of sperm capacitation in mammals, sperm maturation is completed in the spermathecae of Sciara 7 to 9 hours after insemination. It consists of the acquisition of sperm motility and elimination of the electron transparent region of the mitochondrial nebenkern derivative. The electron microscope reveals in mature sperm that the axial filament doublets have changed configuration and consist of a single whorl which encloses the paracrystalline rod. The process by which the major portion of the nebenkern derivative is eliminated occurs in four identifiable stages. Since sperm maturation does not appear to be intrinsically controlled, factors in the spermathecal fluid may play a role in its completion.