Localization of the zona glycoproteins ZPB (ZP3 alpha) and ZPC (ZP3 beta) in the bovine ovary during pre- and postnatal development.

The purpose of this study was to investigate the expression of two zona pellucida gene families ZPB (ZP3 alpha) and ZPC (ZP3 beta). Sections of ovaries from bovine fetuses, calves and cows were labelled with polyclonal antibodies. Immunopositive labelling was found in both the follicle cells and the oocyte. Labelling was dependent on the stage of development. The specific sequence of immunopositive reactions suggests that in the bovine fetus both the ovary and the follicle cells contribute to the production of the zona pellucida during pre- and postnatal development.

Localization of the mRNA encoding the zona protein ZP3 alpha in the porcine ovary, oocyte and embryo by non-radioactive in situ hybridization.

The mRNA of the zona pellucida glycoprotein ZP3 alpha was localized in frozen sections of pig ovaries, isolated oocytes and early embryos by in situ hybridization using biotinylated oligonucleotide probes. In follicles, the distribution of mRNA for ZP3 alpha was correlated with the developmental stage: in primordial and primary follicles, the mRNA was shown to be predominantly localized in the oocyte. In secondary follicles, mRNA was found in both the oocyte and follicle cells. In tertiary and preovulatory follicles, the follicle cells showed distinct staining, whereas the oocyte was labelled weakly. In the early embryo, i.e. 2 days after fertilization, mRNA for ZP3 alpha could not be demonstrated. Our results suggest that, in the pig, the zona pellucida protein ZP3 alpha is synthesized by the oocyte and the follicle cells in sequence. After fertilization, synthesis of ZP3 alpha is terminated.

Early expression of chromogranin A and tyrosine hydroxylase during prenatal development of the bovine adrenal gland.

The present study was undertaken to define the temporal pattern and distribution of cells positive for chromogranin A (CgA) and tyrosine hydroxylase (TH) in various developmental stages of fetal bovine adrenal gland. CgA is an acidic protein, co-stored and co-released with amines and a variety of peptide hormones and neurotransmitters in dense core vesicles of neural and endocrine cells and can be used as a marker for these cells and their malignant counterparts. TH is the rate-limiting enzyme in catecholamine biosynthesis and reflects noradrenergic differentiation. The expression of CgA and TH was examined by immunohistochemistry. CgA immunoreactivity appears first in 35-day-old bovine fetuses. By the end of the second month, CgA-labelled cells are scattered throughout the entire primordium of the adrenal gland, and at a fetal age of 85-91 days most of these cells concentrate in the developing adrenal medulla. From this stage onwards, immunoreactive cells of the marginal zone of the medulla exhibit significantly stronger CgA immunoreaction than the central area. TH immunoreactivity appeared in the adrenal primordium for the first time at the end of the second month of gestation. The distribution pattern of TH-positive cells was similar to that described for CgA, and no significant differences in topographical arrangement between TH- and CgA-positive cells can be detected. The results show that bovine adrenal chromaffin cells express CgA already during their earliest stages of development and prior to TH. The stronger immunoreaction of marginal adrenal medullary cells suggests an adrenalcortical effect of glucocorticoids on the expression of CgA.

Immunocytochemical characterization of porcine zona pellucida during follicular development.

Sections of bovine ovaries fixed in Bouin's fluid or methanol-acetic acid and embedded in paraffin were incubated with chicken polyclonal antibodies to HPLC-purified zona glycoproteins ZP3 alpha and ZP3 beta. Oocytes of primordial follicles as well as of primary follicles showed weak labelling with anti-ZP3 alpha and anti-ZP3 beta. No immunostaining could be observed in the follicle cells. The ZP of primary follicles displayed distinct immunoreactivity for both ZP3 alpha and ZP3 beta. In secondary follicles, distinct labelling with anti-ZP3 beta and weak labelling with anti-ZP3 alpha could be seen in the oocyte. The ZP showed immunoreactivity with antibodies to ZP3 alpha and ZP3 beta. Both antibodies labelled single follicle cells. In tertiary follicles, the oocytes were weakly labelled with anti-ZP3 alpha and anti-ZP3 beta. Some granulosa cells showed staining for ZP3 alpha and ZP3 beta. The ZP displayed strong immunoreactivity for ZP3 beta and ZP3 alpha. Cells of the corona radiata were strongly immunopositive for ZP3 alpha and ZP3 beta. Similar histotopography of immunoreactive cells could be seen in preovulatory follicles. The characteristic pattern observed for the distribution of ZP3 alpha and ZP3 beta strongly suggests that in the porcine ovary both the oocyte and the follicle cells contribute to the synthesis of the ZP, perhaps in sequence.

Development of the bovine duodenum with reference to enterochromaffin cells.

The present light microscopic study describes the morphological changes which occur in the duodenal mucosa of the bovine fetus from the third month of gestation until birth with regard to the sequence of appearance and morphology of enterochromaffin cells. The mucosa of the duodenum is prepared for digestive activity during the first months of development. This becomes obvious in the early differentiation of the fetal intestinal mucosa. The enterocytes in villus epithelium contain glycogen in the early months. These glycogen pools seem to shrink during the following months. They may provide a metabolic reserve for the fetus, which derives all of its nutrition from carbohydrates. Enterochromaffin cells appear in the three month old bovine fetus with two cell-types situated in the epithelial lining of crypts and villi. The density of enterochromaffin cells increases during gestation. There were no findings to indicate that the fetal enterochromaffin cells did not originate from endoderm. The early appearance from enterochromaffin cells (which are the producers of serotonin, 5-hydroxytryptamine) correlates with first gastrointestinal movements, which are known to commence in the first third of gestation, leading us to believe that 5-HT represents the chemical inducer of muscular motility in the developing intestine.

[Fetal development of the omasum of cattle (Bos taurus)]. / Zur fetalen Entwicklung des Blättermagens des Rindes (Bos taurus).

Structures characteristic of the omasum are already present by the third month of development. The fetal omasal laminae exhibit their typical arrangement (1434243414). Muscle fibers from the lamina circularis also extend into the first through third order laminae at this stage. Conical projections and small secondary papillae begin to appear on the laminae as the fetus ages. The lamina muscularis mucosae can be found in all of the laminae by the fourth to fifth month of development. The cuboidal to columnar epithelium of the stratum basale remains largely unaltered throughout the course of fetal development, whereas the stratum superficiale is subjected to a continuous series of changes. Superficial cell layers increase and the glycogen content of the epithelial cells fluctuates significantly.