R.L. Ax

Structural comparisons among glycosaminoglycans to promote an acrosome reaction in bovine spermatozoa.

Bovine epididymal spermatozoa were cultured for 22 hr. in a chemically defined medium containing glycosaminoglycans. Sperm were stained and examined for degree of acrosome reactions and viability after treatment. Viability did not differ significantly among treatments or doses. Highly significant treatment and dose effects resulted for induction of the acrosome reaction. Heparin was the most potent glycosaminoglycan and hyaluronic acid was least effective. Chondroitin sulfate isomers yielded a response intermediate to heparin or hyaluronic acid. These results suggested that the degree of sulfation of the glycosaminoglycans was partly responsible for promoting the acrosome reaction. Treatment of chondroitin sulfate isomers with chondro-4-sulfatase or chondro-6-sulfatase showed that the respective stimulatory effects of chondroitin-4-sulfate and chondroitin-6-sulfate were negated. It was concluded that the ability of glycosaminoglycans to induce the acrosome reaction in bovine spermatozoa in vitro resides in the amino-sugar complex and is enhanced by the presence of a sulfate moiety on the amino-sugar disaccharide.

Proteoglycan from bovine follicular fluid enhances an acrosome reaction in bovine spermatozoa

Abstract Bovine epididymal and ejaculated spermatozoa were incubated for 22 and 9.5 h respectively, in a chemically defined medium. The percentages of sperm exhibiting an acrosome reaction were determined morphologically after fixing and staining specimens. Addition of bovine follicular proteoglycan or chondroitin sulfates ABC significantly increased the incidence of acrosome reaction. The stimulatory effects of the proteoglycan or chondroitin sulfates were negated by exposure to the enzyme chondroitinase ABC. Viability of spermatozoa was not affected by the various experimental treatments. Transmission electron microscopy of spermatozoa showed that vesiculation had occurred between the plasma and outer acrosomal membrane. These results suggest that proteoglycan present in follicular fluid at the time of ovulation may promote the acrosome reaction which precedes the ability of sperm to fertilize an ovum.

Glycosaminoglycans in bovine cumulus-oocyte complexes: morphology and chemistry.

Bovine cumulus-oocyte complexes from small (1-5 mm) follicles were cultured for 24 h in 0.25 ml minimum essential medium supplemented with 10% fetal bovine serum and 20 microCi [3H]glucosamine. Treatment groups consisted of supplementing the culture medium with no hormone (control), 0.5 IU/ml follicle-stimulating hormone (FSH) or 10 mM 8-Br-adenosine cyclic monophosphate (cAMP). After culture, the complexes were fixed for light and scanning electron microscopy. Electron photomicrographs revealed that complexes induced to expand with FSH or cAMP contained a copious glycosaminoglycan (GAG) matrix extending between and around the cumulus cells. Control complexes did not exhibit expansion or an extracellular matrix. The radiolabeled GAG material was isolated for chemical identification. Chemical analyses included: (1) electrophoresis of GAG material, (2) electrophoresis of GAG material after enzyme or nitrous acid treatment, (3) thin-layer chromatography of GAG hydrolysates. The results from electrophoresis showed that the radiolabeled GAG co- migrated with hyaluronic acid. The GAG material was resistant to chondroitinase ABC and nitrous acid degradation but was digested by hyaluronidase. Complexes treated with FSH and cAMP incorporated higher (P less than 0.1 and P less than 0.025 respectively) amounts of [3H]glucosamine into hyaluronic acid than control cultures. Thin-layer chromatography identified the primary amino sugar of the GAG to be glucosamine. These data collectively showed that the radioactive GAG produced by bovine cumulus-oocyte complexes was hyaluronic acid.

Structural variation around prolactin gene linked to quantitative traits in an elite Holstein sire family.

SummaryDigestion of genomic DNA with the restriction endonuclease Avail disclosed a probable insertion deletion of approximately 200 base pairs (bp) near the prolactin gene. Two alleles were apparent as three distinct hybridization patterns. These alleles were statistically associated with quantitative trait loci among sons of one elite Holstein sire family. The favorable genotype was correlated with the presence of a 1.15-kb hybridization band inherited from the sire when genomic DNA was probed with a full-length cDNA for prolactin. Pedigree estimates of genetic merit among genotypes were similar, differing by only 19.3 kg for milk in ancestor merit. Comparisons of genetic estimates for quantitative yield traits in offspring of this heterozygous sire showed significant (P<0.05) differences between homozygous genotypes for predicted difference milk (PDM), predicted difference dollars (PD

Identification of a heparin-binding protein in bovine seminal fluid as tissue inhibitor of metalloproteinases-2

), cheese yield dollars, and protein dollars. The estimated differences between homozygous genotypes for USDA Transmitting Abilities of PDM, PD

Supplementation of progesterone via controlled internal drug release inserts during ovulation synchronization protocols in lactating dairy cows

, Cheese Yield

Purification and characterization of fertility‐associated antigen (FAA) in bovine seminal fluid

and Protein

Proteoglycan production by bovine granulosa cells in vitro occurs in response to fsh

were 282.93 kg,

Pregnancy outcomes in two commercial dairy herds following hormonal scheduling programs

74.35,

Isolation and characterization of seminal fluid proteins that bind heparin.

48.58 and