Monoclonal antibody detection of heparin-binding proteins on sperm corresponds to increased fertility of bulls.

A monoclonal antibody (M1) was produced against seminal fluid heparin-binding proteins (HBP) from a vasectomized bull. In the first part of this study, the presence of HBP in sperm or seminal fluid was determined for 53 bulls with an ELISA using M1. Bulls (8 to 18 per pasture) were bred to 1,114 cows at ratios of 1 bull:25 cows. Bulls with detectable HBP on sperm membranes were 11 percentage points more fertile than bulls with undetectable HBP in sperm membranes. In the second part of this study, three sperm, membrane HBP approximately 30, 24, and 21.5 kDa were identified with Western blots using M1. Santa Gertrudis bulls (n = 64) were bred to 1,354 Santa Gertrudis cows in groups with 2 to 11 bulls. Bulls with those three HBP (Group A) or a single 30-kDa HBP (Group B) in sperm membranes had the greatest fertility, ranging from 74.4 to 89.9% (mean = 81.5%) of the palpated cows that were pregnant. Bulls with the 21.5- and 30-kDa HBP (i.e., the 24-kDa HBP was absent; Group C) had a reduced fertility of 61.3%. Bulls without detectable HBP (Group D) resulted in 41.9% of 186 cows palpated pregnant. Bulls in Groups A and B were more (P < .01) fertile than all other groups. In conclusion, the presence of HBP in sperm membranes was indicative of the fertility potential of bulls.

Estrogen enhances epidermal growth factor-induced DNA synthesis in mammary epithelial cells.

Estradiol (E2) priming (1 nM for 48 h) of normal murine mammary gland epithelial cells significantly increased the response of those cells to epidermal growth factor (EGF)-induced DNA synthesis. The synergism between E2 and EGF was evident in two aspects: After serum-free synchronization for 24 h, more cells entered the S-phase of the cell cycle after E2 priming and when treated with 0.17 nM EGF (13%) than did control cells (1.3%) or cells treated with EGF (4%) or E2 (3.5%) alone; further, the dose of EGF required to elicit maximal response was reduced an order of magnitude in estrogen-primed cells (0.17 nM) compared to controls (1.7 mM). Estrogen alone, however, did not increase DNA synthesis in these cells. Ligand binding studies indicate that these effects of estrogen on proliferating mammary epithelial cells may be explained, at least in part, by a 3.7-fold increase in the number of high affinity EGF-receptors observed in estrogen primed cells (7,300 receptors per cell) compared to estrogen deprived cells (1,960 receptors/cell).

Binding of bovine follicular fluid glycosaminoglycans to fibronectin, laminin and low-density lipoproteins.

Interactions of bovine follicular fluid glycosaminoglycans (GAGs) with extracellular matrix (ECM) components fibronectin and laminin and with low-density lipoproteins (LDL) were examined using affinity chromatography. Glycosaminoglycans from small (diameter less than 5 mm) and large (diameter 11-20 mm) follicles were isolated from follicular fluid. The dermatan sulphate or heparan sulphate from small or large follicles was applied to Fn-, Lm- or LDL-Sepharose columns. Portions of each fraction of the bound or unbound GAG were then subjected to gel filtration h.p.l.c. for quantification. The binding interaction between dermatan sulphate and fibronectin was significantly greater than between heparan sulphate and fibronectin (P less than 0.05); the binding interaction between GAGs from small follicles and fibronectin was significantly greater than between GAGs from large follicles (P less than 0.05). The binding interaction between GAGs from small follicles and laminin was significantly greater than for GAGs from large follicles (P less than 0.05). Dermatan sulphate from small follicles bound to fibronectin (42%), laminin (36%) and LDL (14%) and that from large follicles bound to fibronectin (14%), laminin (23%) and LDL (14%). Heparan sulphate from small follicles bound to fibronectin (17%), laminin (15%) and that from large follicles bound to fibronectin (13%), laminin (10%) and LDL (6%). These results suggest that dermatan sulphate, but not heparan sulphate, from follicles at different stages of development exhibit a varied ability to interact with components of the ECM. Both substances bound to LDL comparably in small amounts.