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Biomedical subjects

G E Seidel

Publications and source records attributed to G E Seidel.

At least 19 recordsLinked to original sources

At least half of capacitated, motile mouse sperm can fertilize zona-free mouse oocytes.

The percentage of individual sperm capable of fertilizing zona pellucida-free mouse oocytes was investigated by placing motile sperm near zona-free oocytes with a micromanipulator. Incubation with one or two capacitated sperm per oocyte resulted in 50% and 70% fertilization, respectively, compared to 88% for cumulus intact (10(5) sperm/ml) and 87% for zona-free (2 x 10(3) sperm/ml) control oocytes. When sperm were treated with .1 microM calcium ionophore A23187 to facilitate the acrosome reaction, fertilization rates for single motile sperm were markedly lower than for capacitated, nontreated single sperm (4% and 35%, respectively). Similar fertilization rates resulted when one sperm was incubated per two ova (4% and 48% per sperm for A23187-treated and controls, respectively). When a lower dose of A23187 (.001 microM) was used to treat sperm, 7% of oocytes incubated with single sperm were fertilized. These experiments demonstrate that at least half of motile, capacitated mouse sperm are capable of fertilizing zona-free mouse oocytes in vitro, and that motile, A23187-treated mouse sperm resulted in poor fertilization rates.

Acrosome

Embryo transfer for cows with reproductive problems.

Surgical embryo recovery and transfer methods were applied to 25 cows and heifers with chronic reproductive problems. In 16 of the 25 animals, abnormalities could not be detected by palpation per rectum and, at laparotomy, the probable cause of low fertility was found to be oviductal obstructions or periovarian adhesions in 6 of them. Four pregnancies were obtained from 3 of these 6 animals. A definitive diagnosis was not obtained through laparotomy in the other 10 animals in this group; however, an abnormal uterine environment associated with senescence may have been responsible for low fertility in the older cows. Eighteen calves were obtained by embryo transfer from 8 of these 10 donor animals. The remaining 9 heifers and cows had lesions detectable by palpation per rectum. Chronic, purulent metritis was found in 2 cows, one of which produced 7 calves by embryo transfer. The other 7 had periovarian adhesions, and 8 calves were obtained from 2 of them. The adhesions in 6 of these 7 cows were attributable to cesarean section. Laparotomy was valuable for defining the extent of the adhesions and establishing a reliable prognosis in some cases.

Animals

Seminal quality, spermatozoal outpost, and testicular changes in growing Holstein bulls.

Nine Holstein bulls, initially between the ages of 17 and 22 mo, awaiting proofs in artificial breeding, were ejaculated each summer for 3 yr, and 8 and 7 survivors for 4 and 5 yr, respectively, to study changes in the same group of bulls and determine the predictive value of early measurements. Semen was collected twice a day, 2 days per wk, for 4 wk. Bulls differed in testicular size, consistency, ejaculate volume, total spermatozoal output, percentage of unstained spermatozoa and abnormal spermatozoa, and in several storage and freezing tests. The largest yearly effects were on testis size and consistency, ejaculate volume, and total output of spermatozoa. The latter increased per bull from 28.3 X 10(9) per wk at 17 to 22 mo of age to 40.9 X 10(9) per wk 4 yr later, representing a high output of spermatozoa with a total of only 20 min of intensive sexual preparation per wk. The correlation between testis size and spermatozoal output was .72. Testicular consistency was indicative of semen quality measured simultaneously as judged by correlations with spermatozoal characteristics ranging from .61 to .95. These characteristics are believed to be representative of those Holstein bulls in artificial breeding which are ejaculated frequently.

Animals

Effect of tris (hydroxymethyl) aminomethane salt of prostaglandin F2alpha on post-thaw motility of bovine spermatozoa.

The effect of the tris (hydroxymethyl) aminomethane salt of prostaglandin F2alpha on the post-thaw motility of bovine spermatozoa was evaluated for two ejaculates from each of five bulls. This salt of prostaglandin F2alpha was added to the glycerol fraction of the extender to provide 37.5, 112.5, or 337.5 microgram of active prostaglandin F2alpha per .5-ml straw. Post-thaw progressive motility was depressed by the addition of the salt of prostaglandin F2alpha. Motilities averaged over both 0 and 2 h of post-thaw incubation were 19.3, 17.8, 13.6, and 5.8% for 0, 37.5, 112.5, and 337.5 microgram.

Animals

In vitro fertilization, culture, and transfer of rabbit ova.

Ovulated rabbit oocytes were fertilized in vitro in chemically defined media supplemented with bovine serum albumin and either cultured up to the expanding blastocyst stage or transferred to recipients after varying periods of culture. Embryos transferred after up to 72 hours of in vitro culture were born as viable young. Oocytes from young virgin does were superior to oocytes from nonvirgin does for the purpose of in vitro fertilization (54% versus 26% fertilized, P less than 0.01). Capacitated sperm from artificially inseminated capacitators resulted in fertilization rates slightly lower than those from naturally mated does (46% versus 57% fertilized, P less than 0.025). Removal of cumulus and corona cells from oocytes with hyaluronidase and repeated aspiration through a fine pipette resulted in lowered fertilization rates (51% versus 73%, P less than 0.025). Linbro Disposo Tray wells were as good as glass tissue-culture dishes for the in vitro mixing of gametes and were more convenient to use. Modified Ham's F10 medium was used to culture the in vitro-fertilized embryos. However, when a modified Brackett's medium was used instead of modified Ham's F10 for the initial 4-hour period after mixing gametes, more oocytes were fertilized (52% versus 28%, P less than 0.01).

Animals

Evaluation of bovine spermatozoal morphologic features after staining or fixation.

Two experiments were conducted to evaluate effects of 3 stains and 2 fixatives on morphologic features of bovine spermatozoa. In experiment 1, the morphologic features of acrosomes of raw and incubated, extended spermatozoa were evaluated after staining with Hancock's Blom's or Wells-Awa's stains or after fixation with buffered glutaraldehyde. Evaluations were done of stained smears by bright field microscopy and of fixed, unstained preparations, by differential interference contrast microscopy, using wet mounts. Raw semen samples from 1st ejaculates of 80 bulls were evaluated. The percentage of spermatozoa with intact acrosomes averaged 83.5% in unstained preparations fixed in glutaraldehyde, compared with averages of 68.1, 74.5, and 67.4% for smears stained with Hancock's, Blom's, or Wells-Awa's procedures (P less than 0.01). From these results, it appeared that procedures for preparing stained smears were detrimental to acrosomes. Although counts for other acrosomal abnormalities differed (P less than 0.01) in each treatment, patterns were inconsistent. With incubated, extended spermatozoa from 57 bulls, glutaraldehyde-fixed, unstained samples had more (55%) intact acrosomes (P less than 0.01) than did samples stained with Hancock's or Blom's procedures (24.0 and 34.7%, respectively, but the former were not significantly different from Wells-Awa-stained smears (49.3% intact acrosomes). In experiment 2, several morphologic characteristics of spermatozoa from 15 1st ejaculates of 7 bulls were evaluated after staining with Hancock's or Blom's stains or after fixation in buffered glutaraldehyde or buffered formal saline fixatives. Higher counts (P less than 0.01) of head abnormalities were found in wet, unstained fixed preparations (4.83, 4.47, 7.87, and 7.93% respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline methods). There were more (P less than 0.05) separated heads on stained, dry smears (1.43, 1.23, 0.47, and 0.47%, respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline procedures). Fixation with buffered glutaraldehyde resulted in higher counts (P less than 0.01) of proximal protoplasmic droplets (2.47, 1.03, 0.67, and 1.43%, respectively, for glutaraldehyde, Hancock's, Blom's, and formol saline procedures). Although not significant, the same trend was observed for distal protoplasmic droplets...

Acrosome