Carlton A. Eddy

Temporal relationships of estrogen, progesterone, and luteinizing hormone levels to ovulation in women and infrahuman primates

These studies were undertaken to ascertain the interval between the estrogen and LH peaks and ovulation in women, rhesus monkeys, and baboons. Estrogen, progesterone, and LH were measured by RIA. Ovulation was documented by visual examination of the ovaries, histology of the corpora lutea, and recovery of ova. The data for human subjects was based on a group of 23 normal women scheduled for surgical sterilization. Blood was drawn daily between 8:30 and 10:30 P.M. beginning on day 10 of the cycle. Surgery was performed 1 to 5 days after the LH peak. The hormonal findings were correlated with the histology of the corpus luteum. The mean interval from the estrogen peak to ovulation was 34 hours, the interval from the estrogen peak to the LH peak was 24 hours, and that from the LH peak to ovulation was 9 hours. In 46 rhesus monkey cycles and in 53 baboon cycles diagnostic serial laparoscopic examinations were initiated following the estrogen peak and repeated every 24 hours until ovulation was confirmed. The mean interval between the estrogen peak and ovulation was 34 hours in the monkey and 41 hours in the baboon. The intervals from the estrogen peaks to the LH peaks were 12 hours in the monkey and 23 hours in the baboon. The intervals from LH peak to ovulation were 22 hours in the monkey and 18 hours in the baboon. Plasma progesterone levels were significantly increased prior to the LH peak in all three species.

Anatomy and physiology of the fallopian tube.

The anatomy and physiology of the human fallopian tube are described and discussed; then, these facts are correlated with clinical considerations as they relate to tubal factor infertility. Anatomically the human oviduct is a tubular, seromuscular organ attached distally to the ovary and proximally to the lateral aspect of the uterine fundus. Its length averages 11-12 cm. The oviduct can be divided into 4 main segments: 1) the infundibulum, whose terminal end contains the tubal ostium; 2) the ampullary region; 3) the isthmic portion; and 4) the intramural or interstitial portion, which is contained in the wall of the uterus. 4 electron micrographs illustrate these areas. Also discussed in this reveiw are the vascular analtomy, the lymphatics, and neuroanatomy of the fallopian tubes. Physiologic functions discussed in this article include the role of the fallopian tube in sperm transport, its part in sperm maintenance and capacitation, and the tubes function in ovum transport, fertilization, and embryo transport. Clinically, the role of the myosalpinx is undetermined, although it may affect tubal motility and ovum transport. The dense adrenergic innervation of the oviductal isthmus, along with the myosalpinx, suggests that innervation may be required for sphincter-like activity, although again no evidence exists for innervation being required in normal reproduction. The mucosa provides nutrients which may or may not be essential to normal reproduction, and its cilia seems uncritical in gamete transport and embryogenesis. Evidence shows that the uterotubal junction and the ampullary-isthmic junction are not necessary for conception (based on success rates of implantation procedures). Reversal of fimbriectomy is the most difficult and up to 1-cm of ampulla may be removed and resected and still maintain fertility.

NOX5 in Human Spermatozoa EXPRESSION, FUNCTION, AND REGULATION

Background: The identity of the reactive oxygen species (ROS)-producing enzyme(s) in human spermatozoa remains uncertain. Results: NOX5 NADPH oxidase, but not NOX1/2/4, is expressed in human spermatozoa and produces superoxide. Inhibition of NOX5 activity reduces spermatozoa motility. Conclusion: NOX5 is the main source of superoxide and is implicated in human spermatozoa motility. Significance: NOX5 might control the numerous ROS-dependent (patho)physiological processes in human spermatozoa. Physiological and pathological processes in spermatozoa involve the production of reactive oxygen species (ROS), but the identity of the ROS-producing enzyme system(s) remains a matter of speculation. We provide the first evidence that NOX5 NADPH oxidase is expressed and functions in human spermatozoa. Immunofluorescence microscopy detected NOX5 protein in both the flagella/neck region and the acrosome. Functionally, spermatozoa exposed to calcium ionophore, phorbol ester, or H2O2 exhibited superoxide anion production, which was blocked by addition of superoxide dismutase, a Ca2+ chelator, or inhibitors of either flavoprotein oxidases (diphenylene iododonium) or NOX enzymes (GKT136901). Consistent with our previous overexpression studies, we found that H2O2-induced superoxide production by primary sperm cells was mediated by the non-receptor tyrosine kinase c-Abl. Moreover, the HV1 proton channel, which was recently implicated in spermatozoa motility, was required for optimal superoxide production by spermatozoa. Immunoprecipitation experiments suggested an interaction among NOX5, c-Abl, and HV1. H2O2 treatment increased the proportion of motile sperm in a NOX5-dependent manner. Statistical analyses showed a pH-dependent correlation between superoxide production and enhanced sperm motility. Collectively, our findings show that NOX5 is a major source of ROS in human spermatozoa and indicate a role for NOX5-dependent ROS generation in human spermatozoa motility.

Antibodies to Chlamydia trachomatis and risk for tubal pregnancy

We performed a case-control study of the effect of exposure to Chlamydia trachomatis on the risk for tubal pregnancy. Sixty women with tubal pregnancies and 60 matched control women with normal second-trimester intrauterine pregnancies were studied. Cases were more likely than controls to have detectable antichlamydial IgG antibodies (82% versus 58%, p less than 0.01) and their mean titers were higher. The prevalence of IgM antibody seropositivity was not different between cases and controls (20% versus 12%, not significant). Compared with women with IgG antibody titers of less than or equal to 1:8 the relative risk for tubal pregnancy for women with titers greater than or equal to 1:128 was 6.6 (95% confidence interval, 2.0 to 21.6). Among women with tubal pregnancies, antichlamydial antibody titers of greater than or equal to 1:128 were significantly associated with pelvic adhesions and inflammatory tubal mucosal damage. Only 17.6% of women with detectable antichlamydial antibody or inflammatory tubal damage reported a history of pelvic inflammatory disease or gonorrhea.

Pelvic Adhesions Following Microsurgical and Macrosurgical Wedge Resection of the Ovaries

The incidence of postoperative adhesion formation following microsurgical and macrosurgical ovarian wedge resection was contrasted in 10 adult female rhesus monkeys. Bilateral wedge resection was performed on day 10 of the luteal phase using microsurgical technique on one ovary and macrosurgical technique contralaterally. Animals were examined 4 weeks postoperatively. Adhesion formation occurred in only one ovary in which microsurgery had been employed (10%). In contrast, adhesion formation followed macroscopic ovarian wedge resection in five ovaries (50%). All adhesions were periovarian, emanating from the suture line on the ovarian surface. Adhesions were most common on the nonovulatory ovary.

Survival of cryopreservation and thawing with all blastomeres intact identifies multicell embryos with superior frozen embryo transfer outcome

OBJECTIVE To assess the impact of survival of cryopreservation and thawing with all blastomeres intact on the outcome of multicell frozen ET. DESIGN Retrospective study. SETTING Academic assisted reproductive technology program. PATIENT(S) One hundred sixteen exclusively multicell frozen ETs in 78 patients. INTERVENTION(S) Frozen ET. MAIN OUTCOME MEASURE(S) Relation of embryonic blastomere survival to the outcome of frozen ET (i.e., pregnancy). RESULT(S) When at least one embryo survived with all blastomeres intact, the total pregnancy rate (biochemical, clinical, or delivered) was 37.7%, the clinical pregnancy rate was 24.6%, and the delivered pregnancy rate was 18.8%. When no embryo survived with all blastomeres intact, the corresponding rates were 10.6%, 8.5%, and 6.4%. The differences in the total pregnancy rate and the clinical pregnancy rate were statistically significant. The delivered pregnancy rates approached statistical significance. CONCLUSION(S) Multicell embryonic survival of cryopreservation and thawing with all blastomeres intact identifies embryos with superior developmental potential.

Cytogenetic and fertility studies of a rheboon, rhesus macaque (Macaca mulatta) × baboon (Papio hamadryas) cross: further support for a single karyotype nomenclature

Historically, two different numbering systems have been used to describe the baboon and macaque karyotypes. However, G-banding studies and, more recently, fluorescence in situ hybridization results have shown that the two karyotypes are virtually identical. To confirm this hypothesis, cytogenetic analysis of an unusual animal, a rheboon, was undertaken. The rheboon reported here, an 18-year-old male, is the only long-term survivor of 26 pregnancies resulting from matings between female baboons (Papio hamadryas) and male rhesus macaques (Macaca mulatta). A G-banded karyotype was prepared from the rheboon and compared with the karyotypes of the two parental species. Spectral karyotyping (SKY) was carried out on the rheboon chromosomes, and the results were compared with SKY studies reported for the baboon and with CISS (chromosome in situ suppression) studies in the rhesus macaque. No differences were detected in any of the rheboons pairs of autosomes, reinforcing the apparent identity of the two parental karyotypes. Based on these results, we argue that a single karyotyping system should be adopted for the two species. Fertility studies were initiated to determine if the rheboon is sterile, as are most hybrid animals. Two semen ejaculates were devoid of sperm. A testicular biopsy revealed hypoplasia of the seminiferous tubules with few Leydig cells and large lumena. Meiotic arrest occurred during meiosis I, resulting in absence of mature spermatozoa. Thus, the testicular and meiotic findings in the rheboon were similar to those observed in other hybrids, even though the parental karyotypes appear identical.

Effect of ovarian endometriosis on ovulation in rabbits

To study the relationship between endometriosis and ovulatory dysfunction, we induced ovarian endometriosis in the rabbit model Adipose tissue was placed in the contralateral ovary as a control. Ovulation was induced with human chorionic gonadotropin, and ovulation points were counted before and after induction of endometriosis. Periovarian adhesions were graded, and ovaries were histologically examined. A significant decrease in the number of ovulation points was observed in ovaries with endometrial tissue (p = 0.001) but not in ovaries that contained adipose tissue (p = 0.095). Periovarian adhesions decreased the number of ovulation points (p less than 0.01) in ovaries that contained adipose or endometrial tissues. Multivariate analysis demonstrated that an increase in adhesion severity was correlated with a decrease in the number of ovulation points (p less than 0.05), but endometrial tissue was not (p = 0.45). We conclude that, in the rabbit model, minimal ovarian endometriosis impairs ovulation primarily through a mechanism related to periovarian adhesions.

The role of cilia in fertility: An evaluation by selective microsurgical modification of the rabbit oviduct

Fertility, ovum transport, and ciliary activity were examined in 35 rabbits after either unilateral microsurgical reversal or double transection (surgical controls) of 1 cm. segments of distal, middle, or proximal ampulla. Fertility was also examined following similar segmental reversal at the level of the distal and proximal isthmus in eight additional rabbits. Reversal of 1 cm. segments of distal, middle, or proximal ampulla effectively prevented pregnancy. In contrast, control animals that underwent double transection without reversal experienced unaltered fertility. Reversal of 1 cm. segments of distal or proximal isthmus was also consistent with normal fertility. Ova placed on the fimbrial surface of oviducts that had undergone ampullary segmental reversal were arrested at the distal anastomosis of the reversed segment. When mechanically displaced into the reversed segment, such ova were transported toward the ovary and were again arrested at the distal anastomosis. Transampullary transport of ova was normal in doubly transected controls. Ova were transported across both anastomosis sites to the ampullary isthmic junction. Studies of ciliary activity confirmed that cilia within reversed segments of ampulla beat in the ovarian direction, counter to that in the rest of the oviduct. Direction of ciliary beat remained unaltered in surgical controls. These experiments demonstrate a critical role of tubal cilia in ovum transport through the rabbit ampulla. In contrast, other mechanisms control ovum transport into and through the tubal isthmus.

Effects of follicular-phase cocaine administration on menstrual and ovarian cyclicity in rhesus monkeys

OBJECTIVE The purpose of this study was to evaluate the effects of daily follicular-phase cocaine administration on menstrual cyclicity, gonadotropin and ovarian steroid levels, ovulation rates, and corpus luteum function in cycling rhesus monkeys. STUDY DESIGN Thirteen normally cycling, drug-naive adult rhesus monkeys were randomized to receive daily intravenous injections of either 4 mg/kg cocaine or an equal volume of saline solution. Treated animals were yoked to pair-fed controls to minimize differences in caloric intake. Daily blood samples were obtained through indwelling catheters for measurement of serum gonadotropin and ovarian steroid levels. Daily vaginal swabs were obtained to determine the onset of menses. Laparoscopy was performed 2 days after the midcycle estrogen peak to check for ovulation. Daily caloric intakes and pretreatment and posttreatment weights were recorded. RESULTS All six of the control monkeys had laparoscopically confirmed ovulation compared with one of seven in the cocaine-treated group (p < 0.004). Cycle length was normal in five of six controls versus one of seven cocaine-treated monkeys. Estradiol levels were significantly higher in the controls versus the cocaine-treated monkeys (p = 0.01) during the first 14 days of the treatment cycle. There were no differences in basal plasma gonadotropin levels between groups. Luteal-phase lengths and luteal-phase plasma progesterone levels were similar in the controls and the single ovulatory cocaine-treated monkey. There were no significant differences in weight change or caloric intake between the two groups. CONCLUSIONS Daily follicular-phase cocaine administration disrupts menstrual cyclicity and folliculogenesis independent of weight loss, caloric intake, and basal gonadotropin levels.