Accumulation of ZP1 and ZPC in Quail Perivitelline Membrane During Follicular Development

The extracellular matrix surrounding the oocyte before ovulation is called perivitelline membrane (PL) in avian species. We previously reported that one of its components, ZPC, is produced in the ovarian granulosa cells by the stimulation of follicle-stimulating hormone and testosterone. Another component, ZP1, is synthesized in the liver, and might be transported to surface of the oocyte in the follicles. These glycoproteins are assembled to form a three-dimensional network of coarse fiber between the granulosa cells and the oocyte. In order to address the mode of PL formation, we investigated the progressive changes in contents of ZP1 and ZPC in PL during the follicular development in quail ovary. Western blot analysis using specific antisera indicated that ZP1 band was first appeared as 97kDa in molecular mass when the granulosa layer was isolated from the fourth largest follicle, and the intensity of the band was dramatically increased after the follicle matured to the third largest one. On the other hand, immunoreactive ZPC appeared as early as in the granulosa layer obtained from the small yellow follicles (SYF), and the intensity of the immunoreactive band increased progressively during follicular development. The immunohistochemical analysis indicated that the immunoreactive ZP1 and ZPC were not detected on the sections of follicular wall of the very small stroma-embedded white follicles. The immunoreactive ZPC was detected in the PL of SYF sections, whereas no detectable ZP1 was seen in the SYF sections. These results demonstrated that the accumulation of ZP1 was not synchronized to that of ZPC in the PL during follicular development. Alternatively, the accumulation of ZPC in the PL was preceded to that of ZP1 during follicular development in quail ovary

Cloning of Perivitelline Membrane Protein; ZP1 in Turkey (Meleagris gallopavo)

In aves, the perivitelline membrane is an extracellular matrix surrounding the oocyte at the time of ovulation, and is composed of several glycoproteins belonging to the zona pellucida (ZP) gene family. ZP1 and ZPC have been identified as the major components of the perivitelline membrane in quail and chicken. In addition, ZPD has been identified as a component of the chicken perivitelline membrane, and is synthesized in ovarian granulosa cells as ZPC. In contrast, ZP1 is synthesized in the liver and transported by blood circulation. In the present study, we isolated complementary DNA of turkey ZP1 from the liver and analyzed the turkey perivitelline membrane and serum using antiserum for quail ZP1. Turkey ZP1 encodes a protein of 943 amino acids containing a signal peptide, glutamine rich repeat, trefoil domain, ZP domain and consensus furin cleavage site, like quail and chicken counterparts. N-glycosylation sites, which have been proposed as important for interaction with sperm, are completely conserved among the birds. Turkey ZP1 shares 93% and 88% sequence identity with quail and chicken ZP1, respectively. In particular, sequence identity of the ZP domain among birds was higher than overall, probably due to the important role of the ZP domain in construction of the perivitelline membrane. Western blot analysis using anti-ZP1 antiserum detected a band which was almost the same molecular mass as quail ZP1, in the lysate of the perivitelline membrane and in the serum of turkey. These results suggest that turkey ZP1 is synthesized in the liver, and is transported by blood circulation to the ovary as reported in quail and chicken

Synthesis of Bioactive HEMA-MPS-CaCl2 Hybrid Gels: Effects of Catalysts in the Sol-Gel Processing on Mechanical Properties and in vitro Hydroxyapatite Formation in a Simulated Body Fluid

We investigated synthetic conditions for the fabrication of bioactive hybrid gels from monomers of 2-hydroxyethylmethacrylate (HEMA) and 3-methacryloxypropyltrimethoxysilane (MPS) in combination with CaCl2, at a starting molar ratio of HEMA: MPS : CaCl2