The human Fallopian tube contains placental protein 5

Placental protein 5 (PP5), a glycoprotein with properties ofa serine protease inhibitor, was found in human Fallopian tubesremoved during proliferative (n = 6) and secretory (n = 6) phasesof the menstrual cycle. The content of PP5 did not differ infimbrial, ampullar and isthmic parts of the tube. In gel filtration,PP5-immunoreactivity from Fallopian tube extracts eluted asone major peak corresponding to a mol. wt of 36 kd. In radioimmunoassay,the dose–response curves of purified placental PP5 andFallopian tubal extracts were parallel. In immunofluorescencestaining, utilizing polyclonal and monoclonal antibodies, PP5was localized to the epithelium of the Fallopian tube, but weakerstaining was observed also in the stroma. As a potential serineprotease inhibitor, PP5 might play a part in the implantationof the fertilized ovum     

Effect of mifepristone on the expression of cytokines in the human Fallopian tube

Cytokines are believed to play a critical role as mediators between the oviduct and the developing embryo. A synchronous development of embryo and endometrium is essential to successful implantation. It seems to be beneficial for embryo development to rest for some time in the Fallopian tube. Expression of cytokines in the human Fallopian tube and the effect of mifepristone were investigated. Fourteen women with regular menstrual cycles and proven fertility, admitted to the hospital for tubal ligation, were randomly allocated to control or treatment groups. Mifepristone 200 mg was given on day LH+2. Surgery was performed on day LH+3 to LH+5. Biopsies were obtained from the ampullar and isthmic regions of the tubes. Expression of interleukin 8 (IL-8), tumour necrosis factor alpha (TNFalpha), transforming growth factor beta (TGFbeta) and leukaemia inhibitory factor (LIF) was analysed using immunohistochemistry. All cytokines except IL-8 showed the same staining intensity both in the ampullar and isthmic region, while IL-8 was more pronounced in the ampullar region in both epithelial and stromal cells. Exposure to mifepristone made the spatial difference in IL-8 disappear and increased the expression of TNFalpha in the epithelium of the isthmus, but had no effect on the expression of TGFbeta1 or LIF. Changes in cytokine expression in the Fallopian tube are likely to influence embryo development, which could contribute to the contraceptive effect of mifepristone.     

Successful conception following Fallopian tube recanalization in infertile patients with a unilateral proximally occluded tube and a contralateral patent tube

BACKGROUND: There are many published case reports of successful conception following transcervical Fallopian tube recanalization (T-FTR) in patients with bilateral proximally occluded Fallopian tubes. However, no serial trials have been published with respect to successful conception following unilateral tubal recanalization in infertile patients with a unilateral proximally occluded tube and a contralateral patent tube. This study was designated to analyse the success rate of T-FTR and the pregnancy rate due to natural fertilization in the lumen of the recanalized tube in these patients. METHODS: We have encountered only 11 patients with this abnormality in our department in the past 10 years. T-FTR with fluoroscopic guidance was performed in these patients, confirmed by at least two hysterosalpingographies to exclude tubal spasm. The uterine catheter devised by us was used during the procedure. RESULTS: All 11 Fallopian tubes were successfully opened by T-FTR. In the six patients who conceived, a preovulatory follicle was demonstrated on the side of the cannulated tube during the conception. The success rate of recanalization, the pregnancy rate due to fertilization in the lumen of the recanalized tube and the successful delivery rate were 100, 55 and 36% respectively. CONCLUSIONS: Our findings suggest that a functional and/or organic disorder in the patent tube resulted in infertility in patients with unilateral proximal tubal obstruction. Our results further show that recanalization of occluded tubes is an effective treatment. Thus, recognition of successful conception following T-FTR in these patients will be beneficial to our clinical approach to this infertile condition.     

Epithelial cells from human fallopian tube in culture

The epithelial cells lining the inner surface of the Fallopian tube influence the reproductive process by both their ciliary and secretory activities. The aim of the present work was to establish a method to culture these cells as a model for more specific studies of their properties. Minor slices of the mucosal ridges were cut and minced extensively using a fine scissor. The resulting pieces were washed once, resuspended in RPMI 1640 with 20% fetal calf serum and seeded in plastic dishes. After 2 days, the medium was replaced with RPMI 1640 containing human albumin, insulin and transferrin. Seven to 10 days later, the cell number had increased 5-8 times in 70% of the cultures. The identity of the cells was assessed after 1-3 weeks in culture. Of the cells, 98% stained positive for the antibody to epithelial cell-specific protein cytokeratin. Electron microscopic studies of the cultures showed epithelial characteristics including cilia, microvilli and intercellular junctions in the form of desmosomes. The cells could be kept in culture for 6-8 weeks. In conclusion, a method to culture epithelial cells from the human Fallopian tube is described. The cells have been identified and they can be kept in culture for 6-8 weeks in quantities sufficient for experimental use.     

Isolated recurrent torsion of the Fallopian tube: case report

We report a rare clinical case of recurrent isolated torsion of the Fallopian tube. An 18 year old woman presented with acute right lower quadrant pain, nausea and vomiting. Torsion of the Fallopian tube was detected by laparoscopy and detorsion was performed. Two years later, a second similar episode of pelvic pain recurred. Having in mind the first episode, diagnosis was facilitated and detorsion was performed in accordance with the patient's wishes. However, the dilemma of ideal management of recurrent cases of torsion of the same tube remains open for discussion. The possibility of torsion of the Fallopian tube and recurrent torsion of the tube, although rare, should be considered in any patient with acute onset of lower abdominal pain.     

Infertility: A simple, non-invasive method of Fallopian tube sperm perfusion

This case report describes a simple non-invasive method of Fallopiantube sperm perfusion using a paediatric Foley catheter, whichdoes not require cannulation of the Fallopian tube and can beperformed on an outpatient basis. The patient achieved an intra-uterinepregnancy with demonstrable fetal heart beats. Subsequently,use of this method in 29 treatment cycles in 14 women with unexplainedinfertility has resulted in six singleton and one twin ongoingclinical pregnancies.     

Immunology: Formation of antisperm antibodies in women treated with Fallopian tube sperm perfusion

Fallopian tube sperm perfusion (FSP) is a combination of ovarianstimulation and intra-uterine insemination using a large volume(4 ml) of inseminate containing 10⁷–10⁸ spermatozoa. Theinseminate will flush the Fallopian tubes and some of it willend up in the pouch of Douglas. In the present study, we haveinvestigated whether the FSP method will result in the formationof serum antisperm antibodies in the female. A total of 184treatment cycles were given to 128 women. The indications fortreatment were: unexplained infertility (n = 35), various infertilitydiagnoses (n = 28) and donor insemination (n = 65). Prior totreatment, 11 (8.6%) women had a positive tray-agglutinationtest (Friberg) and/or a positive immunobead test. After completingone to four treatment cycles, another six (4.7%) women had developedserum antisperm antibodies. The antibodies induced by the treatmentwere of isotype IgM and directed against the tailtip of thespermatozoa. Two of the women, who prior to the treatment hadantisperm antibodies, showed an increase in antibody titre duringtreatment. There was no statistically significant differencein the pregnancy rate between the women with antisperm antibodiesand the women without. In our opinion, the small risk of developingantisperm antibodies is no contra-indication for treating infertilecouples with FSP.     

Physiology: The production of placental protein 14 by human uterine tubal epithelial cells in culture

Cells prepared from the mucosal layer obtained from the fimbrial,proximal ampullary and distal ampullary regions of the humanuterine (Fallopian) tube have been grown in monolayer culture.Immunocytochemistry with anti-cytokeratin, anti-vimentin oranti-CD 45 antibodies indicated that the overwhelming numberof cells were epithelial in nature and were free of fibroblastsand leukocytes. Basal and steroid-stimulated placental protein14 (PP14) production was investigated in tissue obtained fromnine patients undergoing hysterectomy, by addition of oestradioland/or progesterone to confluent cultures. Basal PP14 productionvaried considerably between experiments, probably due to differencesbetween individuals from whom the tissue had been obtained.However, there was no difference in basal PP14 production bycells prepared from the fimbrial, proximal ampullary and distalampullary parts of the tube obtained from the same patient.When total PP14 production by cells obtained from an individualuterine tube was pooled both progesterone and oestradiol significantly(P < 0.05) stimulated the production of PP14 but the effectof progesterone either alone or in the presence of oestradiolwas numerically greater than that of oestradiol alone. ConsideringPP14 production by cells prepared from the different regionsof the tube showed that cells from the fimbrial region weremore responsive to steroid stimulation than cells prepared fromeither the proximal or the distal ampullary regions. All combinationsof hormonal supplementation stimulated PP14 production by cellsfrom the fimbrial region on all days measured (P < 0.05 -P < 0.001). In contrast oestradiol alone had no effect onPP14 production by cells prepared from the proximal and distalampullary parts of the tube on any day measured. Maximum oestradiol-stimulatedPP14 values, expressed as percentages of control, were (mean± SEM) 146 ± 11, 113 ± 7.5 and 108 ±8.5 for cells prepared from the fimbrial, proximal ampullaryand distal ampullary portions of the tube respectively. Maximumprogesterone or progesterone plus oestradiol-stimulated valueswere very similar; those for progesterone plus oestradiol were(mean ± SEM) 151 ± 8.6, 130 ± 6.2 and 118± 10 for cells from the fimbrial, proximal ampullaryand distal ampullary parts of the tube. The results show thata cell culture system has been established which allows theinvestigation of the function of human uterine tubal cells inculture. Using this system we have shown that PP14 productionby these cells is stimulated by steroids and that cells preparedfrom the fimbrial region of the uterine tube appear more responsiveto steroid stimulation than those prepared from other regionsof the tube.     

Andrology: Undetectable expression of genomic progesterone receptor in human spermatozoa

The expression of genomic progesterone receptor in human ejaculatedspermatozoa was investigated. Spermatozoa from 10 fertile donorswho exhibited normal semen parameters were analysed. Indirectimmunofluorescence and an enzyme immunoassay using monoclonalantibodies against genomic progesterone receptor were used.Different types of spermatozoa were studied: fresh, post-swim-up(migrated), capacitated and post-artificial induction of theacrosome reaction by calcium ionophore A23187. Progestin receptor-richT47D human breast cancer cells were used as a positive control,and progestin receptor-poor MDA-MB-231 human breast carcinomacells were used as a negative control. Genomic progesteronereceptor was not detected in fresh, migrated, capacitated andpost-acrosome reaction induction human spermatozoa and MDA-MB-231cells by either indirect immunofluorescence or enzyme immunoassay.However, in T47D cells a mean concentration of 1043.2 ±125.2 fmol genomic progesterone receptor/mg protein was observedby enzyme immunoassay, and indirect immunofluorescence resultswere positive using both flow cytometry and fluorescence microscopy.These findings suggest that the effect of progesterone on humanspermatozoa is not mediated by genomic progesterone receptor.     

Human sperm function in co-culture with human, macaque or bovine oviduct epithelial cell monolayers

Human sperm function was compared in co-culture with monolayersof oviduct epithelial cells (OEC) from three species, human,macaque and bovine. For all species, freeze–thawed andpassaged OEC from females in the periovulatory phase were used.OEC cultured on an extracellular matrix (Matrigel) formed amonolayer which supported human sperm attachment to OEC fromall three species. Spermatozoa in co-culture with OEC from allthree species showed prolonged survival and improved motilitycharacteristics over those cultured in medium alone. This paperdescribes an efficient, repeatable co-culture system for humanspermatozoa which supports sperm attachment to OEC and subsequentlyimproves sperm function over that seen in control medium cultures.Because the improved sperm function in co-culture did not differsignificantly between human and bovine OEC for those attributesstudied, it is proposed that bovine OEC could be used as analternative to human OEC in certain human sperm co-culture studies.Follicular phase bovine OEC from reproductively normal donorsare far more accessible than their human counterparts, thusmaking this co-culture system more widely available for thestudy of human spermatozoa–female tract interactions.     

Uterus and endometrium: The three-dimensional organization of the smooth musculature in the ampulla of the human Fallopian tube: a new morpho-functional model

The three-dimensional organization of the smooth musculaturearound the human ampulla is revealed by means of scanning electronmicroscopy after NaOH maceration and ultrasonic microdissectionof the interstitial connective tissue. The muscular wall ofthe ampulla appears as a continuous network of randomly anastomosedsmooth muscle cell bundles that showed a multidirectional arrangement.The smooth muscle cell bundles modify their orientation alongtheir course, intertwine repeatedly with each other and dichotomize,generating new bundles with a different orientation from thatat the origin. These results demonstrate that the myosalpinxof the human ampulla is not organized into clear cut longitudinally,circularly or spirally arranged layers, as suggested in previouslight microscopy studies. In contrast, the presence of a networkof multidirectional smooth muscle cell bundles revealed in thisstudy suggests that there is no morphological evidence for unidirectionalperistalsis, and that the musculature is probably structurallydesigned to stir rather than push the tubal contents. Thesemorphological findings better explain the random pattern ofpropagation of the contraction waves and the electrical impulsesthrough the smooth musculature of the human ampulla, as postulatedin early experimental physiological studies. Further, they suggesta specific function for the ampullar musculature which may notbe only strictly related to tubal content transport.     

Fallopian tube sperm perfusion (FSP) versus intra-uterine insemination (IUI) in the treatment of unexplained infertility: a prospective randomized study

Prospective randomization of 60 couples with unexplained infertilitywas performed for treatment either with intrauterine insemination(IUI), using a volume of 0.5 ml of the inseminate, or Fallopiantube sperm perfusion (FSP), using a volume of 4 ml of inseminate.The protocols for ovarian stimulation and induction of ovulationwere the same in the two groups. The two groups were similarconcerning age of the female at the start of treatment and thenumber of follicles > 15 mm diameter, the serum oestradiolconcentrations and the endometrial thickness on the day of humanchorionic gonadotrophin (HCG) administration. The mean (±SD)number of motile spermatozoa inseminated was significantly higherin the FSP group than in the IUI group (52 ± 5 x 10⁶and 28 ± 3 x 10⁶ respectively). In the FSP group, 30women were given a total of 52 treatment cycles; 14 clinicalpregnancies occurred in this group, giving a pregnancy rateof 26.9% per cycle and 46.7% per woman. In the IUI group, 28women were given a total of 51 treatment cycles; five clinicalpregnancies occurred, giving a pregnancy rate of 9.8% per cycleand 17.9% per woman. The pregnancy rates per cycle and per womanin the FSP group were significantly higher than in the IUI group(P < 0.05, chi-square test). This study indicates that inthe treatment of couples with unexplained infertility, Fallopiantube sperm perfusion (FSP) is more successful than intra-uterineinsemination (IUI).     

Human Fallopian tubal epithelial cells in vitro:establishment of polarity and potential role of intracellular calcium and extracellular ATP in fluid secretion

A pure population of human Fallopian tubal epithelial cellshas been isolated by enzyme digestion, grown in primary cultureand used to explore the biochemical basis of oviduct fluid secretion.Confluence was achieved in 3–7 days. Immunocytochemicallabelling for cytokeratins indicated that the cells were epithelialin nature and formed extensive desmosomal contacts, producingapolarized layer in culture. By growing the cells on collagen-impregnatedfilters, a small transepithelial electrical potential differencecould be recorded, with the apical side of the cells negativewith respect to the basal side. In addition, the consumptionof glucose and the appearance of lactate were greater on thebasal than on the apical side of the cells. Because intracellularCa²⁺ ([Ca²⁺]1) is well established as a signal transductionagent in epithelial fluid secretion, the effect of a wide rangeof agonists on [Ca²⁺]1 in isolated tubal epithelial cells wasstudied using Fura-2. The only agent which induced a changein [Ca²⁺]1 was extracellular ATP. The transients induced weredependent on both intracellular and extracellular calcium. ATPadded to the basal side of the cells of the polarized layerinduced a transient increase in the potential difference. Thedata are consistent with a potential role for extracellularATP in the regulation of human tubal fluid formation.     

Morphology and fertility after re-anastomosis of the rabbit Fallopian tube with fibrin glue

Morphology and fertility were studied in 20 female New ZealandWhite rabbits after re-anastomosis of the Fallopian tube withfibrin glue and conventional microsurgical techniques. All oviductswere patent postoperatively. No intraperitoneal adhesions wereobserved. There were no significant differences with regardto the number of corpora lutea, implantations and the nidationindex. Morphological studies demonstrated a normal fold patternand ciliation at the side of anastomosis in the sealed oviductsas well as in the sutured oviducts. No intraluniinal fibrindeposits were found. For re-anastomosis of the Fallopian tubewith fibrin glue, splinting is necessary. In some instancesthis may be related to a mucosal trauma. However, under optimalconditions the use of fibrin glue is equivalent to conventionalmicrosurgical anastomosis of the oviduct. For tubocornual, ampullary-ampullaryand isthinic-ampullary anastomoses with luminal disparity, fibringlue seems to be inappropriate.     

Progesterone inhibits insulin-like growth factor binding protein-1 (IGFBP-1) production by explants of the Fallopian tube

The Fallopian tube provides the environment for early embryo growth, a process which is influenced by insulin-like growth factors (IGFs) in the tubal fluid. Whether the bioavailability of tubal IGFs is modulated by locally produced IGF-binding protein (IGFBP-1) is not clear. An explant culture system from human Fallopian tube mucosa was, therefore, developed enabling the potential for IGFBP-1 production by this tissue to be examined directly. Initial characterization of the system established that the explants maintained responsiveness to steroids. Thus, oviduct-specific glycoprotein production, a major product of the oviduct in vivo, continued to be made via an estrogen-sensitive pathway in the culture. The presence of mRNA for IGFBP-1 was established within the explants by the use of quantitative RT-PCR and IGFBP-1 protein was measured by enzyme-linked immunosorbent assay. Although insulin and estradiol had no consistent effect on IGFBP-1, addition of progesterone had a significant inhibitory effect on IGFBP-1 production, both at the mRNA and protein levels. A dose-range of progesterone revealed an incremental inhibitory effect of progesterone on IGFBP-1 output (maximal effect, 25-50 nmol/l), consistent with physiological inhibition of this process during the luteal phase. We suggest that progesterone might, therefore, play a role in controlling the bioavailability of IGFs to the embryo during early development within the Fallopian tube.     

Human fertilization in vivo, with special reference to progression, storage and release of competent spermatozoa

Whilst very large numbers of spermatozoa accumulate in the cervicalmucus and crypts in the hours following coitus, evidence isoffered from species with comparable gonadotrophin surge-to-ovulationintervals to suggest that the cervix is not the site of thefunctional sperm reservoir in women. On the contrary, humansperrnatozaa involved in the events of fertilization may spendthe greater part of the pre-ovulatory interval in the distalportion of the Fallopian tube, that is in the intra-mural segment.This would enable spermatm to be arrested within reach of thesite of fertilization in the ampullary region of the tube, andto be activated and released clwe to the time of ovulation bymeans of a local transfer of high concentrations of ovarianfollicular hormones. The clinical importance of conArming thisproposed role for the intra-mural segment of the tube is slresed,as Is the possibility of specific micm-environments within thelumen of the duct.     

Physiology: Human tubal fluid: formation and composition during vascular perfusion of the Fallopian tube

A vascularly perfused preparation of the Fallopian tube hasbeen developed as a model to study the formation and compositionof human tubal fluid. An artery serving the tube was cannulatedand perfused at a rate of 0.7 ml/min for 1 h with Medium 199supplemented with bovine serum albumin, heparin and antibiotics.A cannula was also inserted into the lumen. Light and scanningelectron micrographs of control and perfused tubes showed thatthe epithelial lining was intact after perfusion. Tubal fluidwas collected in 13 out of 19 experiments. Fluid could alwaysbe collected from patients who were in the follicular phaseof their ovarian cycle. The mean rate of appearance was 48 µl/h.The glucose, lactate and pyruvate concentrations in the tubalfluid, as assessed by fluorescence microanalysis, were 0.53,8.58 and 0.17 mM respectively. There were no correlations betweenmetabolite concentration and the length of perfusion, cannulationtime, patient's age or condition. This technique provides acontrolled method with which to access and examine human tubalfluid and will allow the physiology of both healthy and diseasedtubes to be studied.     

Prostaglandin E2 and F2alpha receptors in the human Fallopian tube before and after mifepristone treatment

Prostaglandins are associated with several reproductive processesin addition to their effects on the vascular system and muscularcontractility. The aim of this study was to gain informationabout the localization of the receptors for PGE₂ (EP1–EP4)and PGF₂ (FP) in the human Fallopian tube and their regulationfollowing treatment with mifepristone. Sixteen healthy fertilewomen received a single dose of 200 mg mifepristone or placeboimmediately after ovulation (LH+2). Laparoscopic sterilizationwas performed on days LH+4 to LH+6. Biopsies were taken fromthe Fallopian tubes bilaterally. The expression of EP1, EP2,EP3, EP4 and FP was analysed using immunohistochemistry andRT–PCR. The co-localization of prostaglandin receptorsand c-kit or e-nos was analysed using confocal microscopy. Theeffect of progesterone, mifepristone and prostaglandin on tubalcontractility was studied. The presence of EP1–EP4 andFP in the Fallopian tube was detected using immunostaining.The receptors were expressed in serosal cells, luminal epithelialcells, and the muscular wall and vessels of the Fallopian tube.Co-localization studies showed that the endothelial cells stainedpositive for EP1–EP4 and FP and that co-localization wasseen for EP4 and c-kit. Decreased contractility was seen afterprogesterone treatment, whereas increased contractility wasseen after PGF₂ and PGE₂ treatment. These data suggest thatboth the transport of the embryo and the communication betweenthe embryo and the Fallopian tube involve the action of prostaglandinsthrough EP and FP receptors in addition to the effect of prostaglandinson the vascular system and muscular contractility.     

Physiology: Morphology and ultrastructure of Fallopian tube epithelium at different stages of the menstrual cycle and menopause

The Fallopian tube has been reported to undergo cyclical changes.However, many studies of tubal ultrastructure have either examinedone segment of the tube only or studied animal oviducts. Theaim of this study was to document in detail the combined morphologicaland ultrastructural features of the epithelial lining alongthe length of the tube in women at different stages of the menstrualcycle. We report an increase in the proportion of ciliated cellsalong the tube, being highest in the fimbriae, but no substantialdifference between the follicular and luteal phases of the menstrualcycle. In the late follicular phase, fragments of cytoplasmicand cellular material were seen in the isthmic lumen but notin the outer tubal segments. Similarly, surface domes and secretorygranules were more prominent in the mid-tube and ampullary sectionsthan in the fimbriae. This surface activity was followed byrelative quiescence in the early/mid luteal phase with reversionto a more active surface but with little secretory activityin the late luteal phase. These findings along the Fallopiantube substantiate the concept of functional differentiationbetween the different segments and necessitate further studiesto determine its clinical relevance.     

Leukaemia inhibitory factor receptor and gp130 in the human Fallopian tube and endometrium before and after mifepristone treatment and in the human preimplantation embryo

Leukaemia inhibitory factor (LIF) is a cytokine, which is associated with reproductive processes such as embryo development and implantation. The objectives of this study were to detect the presence of LIF receptor (LIFR) and glycoprotein 130 (gp 130) in the human Fallopian tube, endometrium and preimplantation embryo and to study the effect of mifepristone on the expression of LIFR and gp130 in the Fallopian tube. Twenty-two healthy fertile women received a single dose of 200 mg mifepristone or placebo immediately after ovulation (LH + 2). Biopsies were obtained from the Fallopian tubes during laparoscopic sterilization once between days LH + 4 and LH + 6 and from endometrium once between days LH + 6 and LH + 8. Preimplantation embryos were received from couples undergoing in vitro fertilization treatment. Immunohistochemistry was used to detect the presence of LIFR and gp130 in the Fallopian tube, endometrium and preimplantation embryo. Real-time PCR was used to study LIFR and gp130 expression in the Fallopian tube and endometrium. LIFR and gp130 were localized in the Fallopian tube, preimplantation embryo and endometrium. LIFR was more abundant in the Fallopian tube than in the endometrium. In the blastocyst, the staining of gp130 was mainly localized in the inner cell mass, whereas LIFR was expressed in all cells. The presence of LIFR and gp130 in the Fallopian tube and preimplantation embryo indicates a role for LIF in communication between the embryo and the Fallopian tube. Mifepristone did not affect the expression of LIFR and gp130 in the Fallopian tube, nor in the endometrium suggesting that progesterone might not be directly involved in the regulation of LIFR or gp130.