Penelope Gaddum-Rosse

Studies on the mucosa of postmenopausal oviducts: surface appearance ciliary activity and the effect of estrogen treatment.

The epithelial lining of the human oviduct is known to be responsive to the fluctuating hormonal levels of the normal menstrual cycle, but its response to the changes in hormonal climate at the time of the menopause is not clearly defined. In this study the oviducts of nine postmenopausal patients were obtained at the time of abdominal hysterectomy, and the lining epithelium was studied by scanning electron microscopy. The activity of cilia on the fresh tissue was assessed by their ability to transport particulate matter applied to the epithelial surface. The fimbriae of oviducts from women who had received little or no estrogen treatment before surgery showed a significant deciliation of the epithelium, compared with specimens from premenopausal patients, and even showed some sloughing of cells from the surface. The secretory cells appeared inactive. However, the specimens from patients who had been treated with estrogen for periods of 1 year or more showed a remarkable maintenance of the epithelium, with the proportion of ciliated cells remaining almost as high as in premenopausal oviducts, even as late as 25 years after the menopause. The ampullar and isthmic portions showed less obvious changes. Cilia in oviducts from the former group (short-term or no treatment) were incapable of transporting 15-mum microspheres or lycopodium spores applied to the epithelial surface, whereas the oviductal cilia obtained from patients under long-term estrogen therapy showed efficient transport of particulate matter. The results are discussed in relation to earlier conflicting reports on the postmenopausal oviduct.

In vitro fertilization in the rat: observations on living eggs**Supported by USPHS grant HD-13075 from the National Institutes of Health and grant GA-PS-7915 from the Rockefeller Foundation. D. E. B. was supported by Developmental Biology Training grant 07183 from the National Institutes of Health.

The purpose of this study was to observe and record some of the key events during in vitro fertilization in the rat. Freshly ovulated eggs were incubated with epididymal spermatozoa at 37 degrees C and removed 4.5 to 7.5 hours later for microscopic examination. The head of the fertilizing spermatozoon penetrated the zona pellucida with its long axis perpendicular to the zona: this orientation was maintained during subsequent incorporation into the vitellus. Sperm motility was drastically reduced soon after sperm-egg fusion. Simultaneously, the flagellum, most of which was still outside the zona, assumed a characteristic curved posture. Time-lapse cinematography demonstrated that the vitellus frequently underwent surface movements during the tail incorporation process, suggesting that its cortex was undergoing significant changes. This study presents the first long-term observations on the fertilization of living rat eggs in vitro.

Studies on Sperm Motility by Laser-Light Scattering

Measurement of sperm motility has received considerable attention of many investigators since the earliest observations of Leeuwenhoek. However, sperm motility has often been judged subjectively by an individual. Therefore, a suitable definition for the measurement of sperm motility continues to confuse the observer. Many methods of measurement have been developed in recent years (1,2). Essentially, these techniques can be divided into two categories depending upon the objectives of the study, which may be associated with the mechanism of flagellation of single cells or may be concerned with the evaluation of relevant statistical parameters which describe the quality of sperm motility. In the studies of effects of exogenous factors on sperm motility and clinical evaluations of semen quality, the microphoto-graphic technique for studying single cells becomes cumbersome due to its inherent disadvantage of being time-consuming. This paper summarizes some of our recent studies of sperm motility by laser-light scattering. Results of these measurements show that this simple and objective method has considerable potential for both basic research and clinical use in biomedical studies.

Laser light-scattering study of the effect of washing on sperm motility**Supported in part by grant HD-12629 to Dr. Wylie I. Lee and grant HD-13075 to Dr. Penelope Gaddum-Rosse, from the National Institutes of Health. Presented in part at the Thirty-Eighth Annual Meeting of The American Fertility Society, March 20 to 24, 1982, Las Vegas, Nevada.

The effect of washing on human sperm motility was measured by means of dynamic laser light-scattering spectroscopy. Semen samples from 24 fertile donors were diluted with Biggers, Whitten and Whittingham (BWW) medium and subsequently centrifuged at one of the following forces: 235 x g, 325 x g, 400 x g, 470 x g, 500 x g, 600 x g, and 800 x g. The duration of centrifugation was 8 minutes for the first wash, 6 minutes for the second wash, and 3 minutes for the third wash. Sperm motility was evaluated in terms of the root mean square swimming speed of the spermatozoa and the mean migration rate of washed spermatozoa in estrous bovine cervical mucus (BCM). It was found that sperm motility and viability were improved when semen samples were washed at 235 x g, even after three washes. However, washing at forces of 600 x g or more reduced sperm motility and also their ability to penetrate cervical mucus in vitro. Repeated washing at forces between 300 x g and 500 x g had little deleterious effect on sperm motility.

Sperm penetration into cervical mucus in vitro. III. effect of freezing on estrous bovine cervical mucus**Supported in part by grants HD-03752 and HD-12629 from the National Institutes of Health.††Presented at the Thirty-seventh Annual Meeting of The American Fertility Society, Atlanta Hilton, Atlanta, Georgia, March 14 to 18, 1981.

The influence of the storages period on estrous bovine cervical mucus after it was stored in the freezing compartment of the laboratory refrigerator was evaluated by an in vitro sperm penetration test with human spermatozoa, laser light-scattering, and a spinnbarkeit test. Data obtained from the sperm penetration test were analyzed by a mathematical model that correlates the sperm motility with the sperm transport rate and the penetrability of the mucus. The tests showed that estrous bovine cervical mucus can be stored for up to 4 weeks at -12 degrees C without a change in its physical properties. The results of this study strengthen the suggestion that bovine mucus could be employed as a substitute for human cervical mucus.

Sperm Penetration into Cervical Mucus In Vitro. II. Human Spermatozoa in Bovine Mucus**Supported by United States Public Health Service Grant HD-03752 and by Contract HD4-2826 from the National Institutes of Health.

Human spermatozoa pentrate estrous bovine cervical mucus readily in vitro and maintain good motility and viability for a number of hours. They show pronounced unidirectional motion in mucus that has been aligned linearly. Data from tube preparations indicate that human spermatozoa from a given ejaculate travel more rapidly in estrous bovine mucus than in human midcyle mucus. They are prevented from penetrating luteal phase bovine mucus. The results are discussed in relation to a model of the molecular structure of cervical mucus, derived from laser light-scattering spectroscopy. In addition, it is suggested that bovine cervical mucus could be developed as a possible substitute for human cervical mucus in cases of infertility due to deficient endogenous mucus.

Sperm Penetration into Cervical Mucus In Vitro. I. Comparative Studies**Supported by United States Public Health Service Grant HD-03752 and by Contract HD4-2826 from the National Institutes of Health.

Comparative studies have been carried out on the behavior of human and bovine spermatozoa toward homologous cervical mucus in vitro. In both cases the degree of sperm penetration and the pattern of sperm motility were influenced in a characteristic fashion by prior manipulation of the mucus: the most rapid and extensive penetration, and pronounced unidirectional motion, were seen in mucus that had been aligned linearly. By contrast, spermatozoa from rabbits, guinea pigs, rats, and mice were largely prevented from entering either midcycle human or estrous bovine cervical mucus, regardless of its physical arrangement. The observations on sperm motility patterns and the degree of penetration are discussed in relation to a model of the molecular arrangement of cervical mucus, derived in our laboratory from laser light-scattering spectroscopy.