Germ Cell Clusters Research Articles

An in vitro model of gonad differentiation in the chick embryo. Roller cultures in gas permeable biofoil bags.

Embryonic gonads of 6 1/2 to 12 days old chick embryos were enzymatically dissociated. The cell suspensions were cultured in small gas permeable bags of foil (Biofolie Heraeus) in a roller culture apparatus. The cells formed multiple small aggregates, in which sex specific differences developed within two days. In cell suspensions of embryonic testes smooth spheric aggregates formed with well delineated testicular cords in the center and a tunica albuginea-like mesenchymal layer at the outside. Most of the male germ cells were incorporated in the central cords. A number of germ cells were barred from entering the cords by the tunica albuginea-like mesenchymal layer and populated the outer surface of the aggregates. The aggregates of left ovary were irregular in shape and characterized by clusters of germ cells residing in an outer cortical zone. The aggregates of the right ovary, which regresses in vivo, showed poor growth and did not differentiate, thus, indicating that the suppression of right ovary was not removed in culture. In the roller cultures of dissociated embryonic gonads male and female morphogenesis was mimicked in a reproducible manner, so that the system can be used for further experimental studies of gonadal development.

Ultrastructure of oogenesis in the holopelagic polychaetes Rhynchonerella angelini and Alciopa reynaudii (Polychaeta: Alciopidae)

Ultrastructural features of oogenesis were examined in the pelagic polychaetes Rhynchonerella angelini and Alciopa reynaudii which were collected from Bahamian waters by a manned submersible during 1979 and 1980. No definitive ovary was detected in either species. Oogonia are released into the coelom as packets of cells, where they undergo mitotic division while surrounded by an envelope of sheath cells. Cytokinesis is incomplete, resulting in intercellular bridges between oogonia. Oocytes undergo early stages of meiosis characterized by the presence of synapsed chromosomes, followed by a period of rapid cytoplasmic and nuclear growth. Oocytes are released from the packets in the early vitellogenic phase into the coelom, where they undergo yolk synthesis as solitary coelomic cells. Vitellogenesis includes both autosynthetic and heterosynthetic processes. Autosynthesis involves the fusion of secretory vesicles formed by the combined activity of the rough endoplasmic reticulum and Golgi complex, with convoluted electron-dense tubular bodies of unknown origin. Heterosynthesis involves the intense uptake of exogenous precursors through endocytosis and their fusion into nascent yolk bodies which, in turn, are presumed to fuse with autosynthetically-derived yolk bodies. No nutrient stores were detected in somatic tissues. Early and middle stages of vitellogenic oocytes were absent from the coelom. This absence combined with the high level of endocytotic activity suggests that vitellogenesis occurs rapidly. These features, in combination with the presence of an exceptionally thin body wall and gut, might serve as related adaptations for predator avoidance by the maintenance of relatively low tissue-density. Alciopid, phyllodocid, and nereid polychaetes share some common reproductive features including the presence of “dispersed” ovaries, clusters of syncytial germ cells which undergo meiosis while enveloped by somatic cells and the release of oocytes from the clusters prior to vitellogenesis.

The flea ovary: Ultrastructure and analysis of cell clusters

Panoistic ovarioles are found in the order of fleas (Siphonaptera). Only in some species of the Hystrichopsylloidea do polytrophic meroistic ovaries occur. No stem cells and no dividing cystocytes are found in female imagines of Hystrichopsylla talpae. However, each germ cell cluster consists of 32 cells which are generated by five mitotic cycles during the pupal stage. One of the cells containing five intercellular bridges becomes the oocyte, the others serve as nurse cells. Thus, germ cell cluster formation follows the 2 n -rule. However, no polyfusome is found and nurse cells do not form a rosette. Furthermore, nurse cells remain small and show the same ultrastructural characters as the oocytes, which became distinguishable from nurse cells only by their enhanced growth during pre-vitellogenesis. The first phase of pre-vitellogenesis is dominated by the production of an unknown cytoplasmatic component, consisting of spherical particles, clearly distinguishable from ribosomes by diameter and contrast. The next phase is characterized by a tremendous increase in the production of ribosomes. During this second phase another cytoplasmic component consisting of ball-like structures becomes prominent. During pre-vitellogenesis, germ cell nuclei undergo a pronounced structural change in which, finally, numerous extranucleolar particles predominate. Thus, H. talpae has a polytrophic meroistic ovary, but its oocyte genomes behave panoistically.

Morphology, ultrastructure, and germ cell cluster formation in ovarioles of aphids.

The structure of aphid ovaries, including ovipare and virginopare morphs of five species, was investigated by light and electron microscopy. Aphids contain telotrophic meroistic ovarioles. The amount and distribution of cytoplasmic components of nurse cells, nutritive cords, and young oocytes are nearly identical to those known from scale insects and heteropterans. Each ovariole has a constant number of nurse cells and oocytes. In ovaries of ovipare morphs, the nurse cell nuclei enlarge by endomitosis (n = 28 n-210 n), whereas in virginopare morphs the nurse cell nuclei remain small (n = 22 n-24 n). Furthermore, in virginoparae the previtellogenic growth of oocytes is highly reduced, and vitellogenesis and chorionogenesis are blocked totally. Embryogenesis starts immediately after the shortened previtellogenic growth. In each ovariole, all germ cell descendants belong to one germ cell cluster that follows the 2n rule. The cluster normally contains 25 = (32) cells, but other mostly smaller numbers also occur. In contrast to polytrophic meroistic ovarioles, more than one cell of each cluster will develop into an oocyte. In Drepanosiphum platanoides, 16 (2n-1 ) nurse cells and 16 (2n-1 ) oocytes exist in each cluster, whereas, in Metopolophium dirhodum, 8 (2n-2 ) oocytes and 24 (2n-1 + 2n-2 ) nurse cells are normally found. In many ovarioles of Macrosiphum rosae, 21 nurse cells nourish 11 oocytes. Models of germ cell cluster formation in aphid ovaries are discussed.

Separation of mouse spermatogenic cells by sedimentation velocity: A morphological characterization

A method utilizing sequential enzymatic incubation in collagenase (1 mg/ml) and trypsin (2.5 mg/ml) has been developed for the dissociation of the seminiferous epithelium. A significant advantage of this method is that, following collagenase incubation and washings in an enriched Krebs-Ringer bicarbonate buffer solution, isolated seminiferous tubules are obtained which are free of interstitial cells. The “purified” seminiferous epithelium is then dissociated with trypsin. A further advantage of this dissociation technique has been a reduction in the number of symplasts (multinucleate cells) which form by the opening up of the intercellular bridges that occur between synchronously differentiating clusters of germ cells. Both the elimination of the interstitial cells and the reduction in the number of symplasts have made possible the recovery of more highly enriched germ cell fractions. The homogeneity of the cell fractions was determined by light and electron microscopy. Integrity of the isolated cells was verified by Trypan blue exclusion and measurement of oxygen consumption.

The embryology of Lytta viridana LeConte (Coleoptera: Meloidae). VI. The appendiculate, 72-h embryo

The embryology of L. viridana is continued to the 72-h stage, when a little over one-quarter of the embryonic period has elapsed. The origin of the segmental appendages, further development of the coelomic sacs and enteron rudiments, formation of the nerve ganglia, appearance of tracheal invaginations, migration of the germ cells, and secretion of the serosal cuticle are described. The embryonic origin of the labrum is preoral and preantennal, that of the antennae and deutocerebrum paraoral. The tritocerebrum is formed in a well-defined intercalary segment. The ventral musculature of the stomodaeum arises from intercalary mesoderm, the dorsal and lateral musculature from labral mesoderm. The outer sheaths of the Malpighian tubules are formed from mesoderm of the 10th abdominal segment, the musculature of the proctodaeum from the 11th segment. The pleuropodia differentiate rapidly and begin secreting. The germ cell rudiment subdivides into three to six pairs of germ cell clusters, which become arranged segmentally.