On the basis of structural observations bovine oocytes were grouped into four successive classed: 0, those before the luteinizing hormone (LH) surge; 1, those up to 8 h following the LH peak level; 2, those between 8 and 19 h after the LH peak level; and 3, those between 19 h after the LH peak level and ovulation.
Oocytes in class 0 had mitochondria located in a generally peripheral position. Interior to the mitochondria were elements of rough endoplasmic reticulum (RER) and numerous membrane‐bound vesicles which bore ribosome‐like particles on their outer surface. The first visible changesater the LH peak level as seen in class 1 were the formation of the periviteline space with loss of contact between the cumulus cells and the oocyte, and ruffing of the nuclear envelope. These changes were followed b the resumption of meiosis as defined by germinal‐vesicle breakdown (GVBD), the disappearance of RER, and the formation fo clusters of mitochondria in association with lipid droplets and elementrs of smooth endolasmic reticulum (SER).
The period between 8 and 19 h following LH peak level (class 2) was characterized by intensive clustering of mitochoncria in association with lipid droplets and elements of SER, conversion of lipid, fusion of vesicles, and the appearance of ribosomes in the cytoplasm. During the final stage (class 3), the polar body was extruded, the mitochondria dispersed, and the majority of the organelles became located toward the center of the cell. The relatively organelle‐free cortical region contained cortical granules immediately adjacent to the plasma membrane together with aggregates of tubular SER. The structural changes are discussed in the context of follicular steroidogenesis and oocyte developmental competence.
The structural changes that characterize primary, secondary and tertiary atresia in sheep Graafian follicles have been studied by means of histological, histochemical and ultrastructural techniques. In primary atresia vacuoles representing swollen endoplasmic reticulum are prominent along the antral border together with disorganized granulosa cells containing pyknotic nuclei. Phagocytic cells, which increase in number as atresia progresses, were seen within the membrana granulosa and are considered to be transformed granulosa cells. Even in follicles classified as nonatretic, a few antral vacuoles and occasional pyknotic nuclei are present. During secondary atresia there is a large increase in the number of cells with pyknotic nuclei; many of these nuclei had been extruded and had fused to form the characteristic Feulgen-positive atretic bodies found along the edge of the antral cavity. These bodies usually have a diameter of up to 15 mum but occasionally reached as much as 400 mum. A second area of degeneration is frequently present in the membrana granulosa, two or three cell layers from the basal lamina, and it is at this level that exfoliation of granulosa cells occurs in tertiary atresia. In contrast to the membrana granulosa, there are during secondary atresia, only slight indications of degeneration in the cumulus. In tertiary atresia the membrana granulosa is highly disorganized; the atretic bodies are often fewer in number than at earlier stages. The basal lamina remains essentially intact. It is at this stage that the first clear signs of degeneration occur in the theca interna. Despite some disintegration of the cumulus, the integrity of the oocyte is maintained and its nucleus remains vesicular. Changes in the thecal microcirculation may plan a key role in atresia: adjacent to the basal lamina of non-atretic follicles, there is a well-developed capillary network which is significantly reduced as atresia progresses.
Embryonic development entails a well defined temporal and spatial programme of gene expression, which may be influenced by active chromosomal domains. These chromosomal domains can be detected using transgenes which integrate randomly throughout the genome, as their expression can be affected by chromosomal position. Position effects are probably exerted most strongly on transgenes that do not contain strong promoters, enhancers or other modulating sequences. Here we have systematically explored position effects using a transgene with the weak herpes-simplex-virus thymidine-kinase promoter, linked to the readily visualized lacZ indicator gene (HSV-TK-lacZ). Each transgenic fetus with detectable expression displayed a unique lacZ staining pattern. Thus expression of this construct is apparently dictated entirely by its chromosomal position, without any construct specificity. Furthermore the transgene is faithfully transmitted to subsequent generations, allowing for systematic mapping of changes in expression during development and in adult life. These results demonstrate that transgenes can indeed be powerful tools to probe the genome for active chromosomal regions, with the potential for identifying endogenous genes involved in organogenesis and pattern formation.
Pig oocytes were examined at hourly intervals after stimulation with hCG. Meiosis was resumed between 20 and 30 h after hCG. This coincided with a decline in the number of mitochondria and evidence is presented which indicates that this was due to fusion. The number of lipid droplets increased and the volume fraction of large vesicles decreased. Both these organelles maintained a close spatial relationship with the endoplasmic reticulum (ER). Mitochondria were clustered at the periphery of the cell before hCG injection but dispersed with maturation. The volume occupied by large vesicles, 'protein bodies' and Golgi also decreased at the edge of the oocyte with the progression of maturation.
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