Cells Of Reproductive Tract Research Articles

Female Sex Hormone Regulation of Tenofovir-diphosphate in Human Female Reproductive Tract (FRT) Cells in Culture

AIDS Research and Human RetrovirusesVol. 30, No. S1 Novel Formulations, Agents and MicrobicidesFemale Sex Hormone Regulation of Tenofovir-diphosphate in Human Female Reproductive Tract (FRT) Cells in CultureZheng Shen, John V. Fahey, Marta Rodriguez-Garcia, Jack E. Bodwell, Angela D.M. Kashuba, and Charles R. WiraZheng ShenGeisel School of Medicine at Dartmouth, Physiology and Neurobiology, Lebanon, NH, United StatesSearch for more papers by this author, John V. FaheyGeisel School of Medicine at Dartmouth, Physiology and Neurobiology, Lebanon, NH, United StatesSearch for more papers by this author, Marta Rodriguez-GarciaGeisel School of Medicine at Dartmouth, Physiology and Neurobiology, Lebanon, NH, United StatesSearch for more papers by this author, Jack E. BodwellGeisel School of Medicine at Dartmouth, Physiology and Neurobiology, Lebanon, NH, United StatesSearch for more papers by this author, Angela D.M. KashubaUNC Eshelman School of Pharmacy, The Division of Pharmacotherapy and Experimental Therapeutics, Chapel Hill, NC, United StatesSearch for more papers by this author, and Charles R. WiraGeisel School of Medicine at Dartmouth, Physiology and Neurobiology, Lebanon, NH, United StatesSearch for more papers by this authorPublished Online:30 Oct 2014https://doi.org/10.1089/aid.2014.5306.abstractAboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View article"Female Sex Hormone Regulation of Tenofovir-diphosphate in Human Female Reproductive Tract (FRT) Cells in Culture." AIDS Research and Human Retroviruses, 30(S1), pp. A149–A150FiguresReferencesRelatedDetailsCited byA Translational Pharmacology Approach to Predicting Outcomes of Preexposure Prophylaxis Against HIV in Men and Women Using Tenofovir Disoproxil Fumarate With or Without Emtricitabine24 February 2016 | Journal of Infectious Diseases, Vol. 214, No. 1Pharmacokinetics of antiretrovirals in mucosal tissue22 March 2015 | Expert Opinion on Drug Metabolism & Toxicology, Vol. 11, No. 6 Volume 30Issue S1Oct 2014 InformationCopyright 2014, Mary Ann Liebert, Inc.To cite this article:Zheng Shen, John V. Fahey, Marta Rodriguez-Garcia, Jack E. Bodwell, Angela D.M. Kashuba, and Charles R. Wira.Female Sex Hormone Regulation of Tenofovir-diphosphate in Human Female Reproductive Tract (FRT) Cells in Culture.AIDS Research and Human Retroviruses.Oct 2014.A149-A150.http://doi.org/10.1089/aid.2014.5306.abstractPublished in Volume: 30 Issue S1: October 30, 2014PDF download

Sex hormones regulate tenofovir-diphosphate in female reproductive tract cells in culture.

The conflicting results of recent pre-exposure prophylaxis (PrEP) trials utilizing tenofovir (TFV) to prevent HIV infection in women led us to evaluate the accumulation of intracellular TFV-diphosphate (TFV-DP) in cells from the female reproductive tract (FRT) and whether sex hormones influence the presence of TFV-DP in these cells. Following incubation with TFV, isolated epithelial cells, fibroblasts, CD4+ T cells and CD14+ cells from the FRT as well as blood CD4+ T cells and monocyte-derived macrophages convert TFV to TFV-DP. Unexpectedly, we found that TFV-DP concentrations (fmol/million cells) vary significantly with the cell type analyzed and the site within the FRT. Epithelial cells had 5-fold higher TFV-DP concentrations than fibroblasts; endometrial epithelial cells had higher TFV-DP concentrations than cells from the ectocervix. Epithelial cells had 125-fold higher TFV-DP concentrations than FRT CD4+ T cells, which were comparable to that measured in peripheral blood CD4+ T cells. These findings suggest the existence of a TFV-DP gradient in the FRT where epithelial cells > fibroblasts > CD4+ T cells and macrophages. In other studies, estradiol increased TFV-DP concentrations in endometrial and endocervical/ectocervical epithelial cells, but had no effect on fibroblasts or CD4+ T cells from FRT tissues. In contrast, progesterone alone and in combination with estradiol decreased TFV-DP concentrations in FRT CD4+ T cells. Our results suggest that epithelial cells and fibroblasts are a repository of TFV-DP that is under hormonal control. These cells might act either as a sink to decrease TFV availability to CD4+ T cells and macrophages in the FRT, or upon conversion of TFV-DP to TFV increase TFV availability to HIV-target cells. In summary, these results indicate that intracellular TFV-DP varies with cell type and location in the FRT and demonstrate that estradiol and/or progesterone regulate the intracellular concentrations of TFV-DP in FRT epithelial cells and CD4+ T cells.

Extrapineal melatonin: sources, regulation, and potential functions.

Endogenous melatonin is synthesized from tryptophan via 5-hydroxytryptamine. It is considered an indoleamine from a biochemical point of view because the melatonin molecule contains a substituted indolic ring with an amino group. The circadian production of melatonin by the pineal gland explains its chronobiotic influence on organismal activity, including the endocrine and non-endocrine rhythms. Other functions of melatonin, including its antioxidant and anti-inflammatory properties, its genomic effects, and its capacity to modulate mitochondrial homeostasis, are linked to the redox status of cells and tissues. With the aid of specific melatonin antibodies, the presence of melatonin has been detected in multiple extrapineal tissues including the brain, retina, lens, cochlea, Harderian gland, airway epithelium, skin, gastrointestinal tract, liver, kidney, thyroid, pancreas, thymus, spleen, immune system cells, carotid body, reproductive tract, and endothelial cells. In most of these tissues, the melatonin-synthesizing enzymes have been identified. Melatonin is present in essentially all biological fluids including cerebrospinal fluid, saliva, bile, synovial fluid, amniotic fluid, and breast milk. In several of these fluids, melatonin concentrations exceed those in the blood. The importance of the continual availability of melatonin at the cellular level is important for its physiological regulation of cell homeostasis, and may be relevant to its therapeutic applications. Because of this, it is essential to compile information related to its peripheral production and regulation of this ubiquitously acting indoleamine. Thus, this review emphasizes the presence of melatonin in extrapineal organs, tissues, and fluids of mammals including humans.

A role of junction-mediated interactions in cells of the male reproductive tract: impact of prenatal, neonatal, and prepubertal exposure to anti-androgens on adult reproduction.

Male sexual development and male reproductive functions are dependent on the normal action of androgens, and an unbalanced ratio of the active androgens can lead to varying degrees of structural and functional abnormalities within the reproductive organs. Endocrine balance can be disturbed by environmental and pharmaceutical anti-androgens (i.e. vinclozolin, phthalates, procymidone, and flutamide) that antagonize normal androgen action. Such chemical compounds enter the cell, bind to the receptor and inactivate transcription leading to disruption of androgen-mediated signaling. Assembling and functioning of cell junctions in hormone-dependent tissues, such as testis, epididymis and prostate appeared to be controlled by steroid hormones, predominantly by androgens. This review presents recent findings on the tight junction proteins mainly responsible for normal functioning of the barrier within the testis, epididymis and prostate, anchoring junction proteins that play a crucial role in normal cell-cell adhesion, and gap junction proteins through which intercellular communication takes place in the male reproductive tract. The review gives examples of animal models that are used in endocrine disruption studies with a focus on the author's own data from studies in the pig.

Sex Hormones Regulate Tenofovir-Diphosphate in Female Reproductive Tract Cells in Culture

Sex Hormones Regulate Tenofovir-Diphosphate in Female Reproductive Tract Cells in Culture

Effect of Prediabetes on Membrane Bicarbonate Transporters in Testis and Epididymis

The formation of competent spermatozoa is a complex event that depends on the establishment of adequate environments throughout the male reproductive tract. This includes the control of bicarbonate (HCO₃⁻) concentration, which plays an essential role in the maintenance of extracellular and intracellular pH (pH(i)) values. Diabetes mellitus alters pHi regulation in mammalian cells, mainly by altering the activity of ion transporters, particularly HCO₃⁻-dependent mechanisms. Yet, little is known about the effects of this pathology and its prodromal stage, prediabetes, on the membrane transport mechanisms of male reproductive tract cells. Herein, we analyzed protein and mRNA levels of the most relevant HCO₃⁻ transporters of the SLC4 family [anion exchanger 2 (AE2), Na⁺-driven Cl⁻/HCO₃⁻ exchanger (NDCBE), electrogenic Na⁺/HCO₃⁻ cotransporter 1 (NBCe1), electroneutral Na⁺/HCO₃⁻ cotransporter 1 (NBCn1)] in the testis and epididymis of a prediabetic animal model. Firstly, we identified the HCO₃⁻ transporters of the SLC4 family, in both testicular and epididymal tissue. Secondly, although no alterations were detected in protein expression, mRNA levels of NBCe1, NBCn1 and NDCBE were significantly increased in the testis of prediabetic rats. On the other hand, in the epididymis, prediabetes caused an increase of AE2 and a decrease of NDCBE protein levels. These alterations may be translated into changes of HCO₃⁻ transepithelial epididymal fluxes in vivo, which may represent a threat for sperm survival. Moreover, these results provide evidence of the molecular mechanism that may be responsible for the significant increase in abnormal sperm morphology already reported in prediabetic rats.

IFN-ε is constitutively expressed by cells of the reproductive tract and is inefficiently secreted by fibroblasts and cell lines.

Type-I interferons (IFNs) form a large family of cytokines that primarily act to control the early development of viral infections. Typical type-I IFN genes, such as those encoding IFN-α or IFN-β are upregulated by viral infection in many cell types. In contrast, the gene encoding IFN-ε was reported to be constitutively expressed by cells of the female reproductive tract and to contribute to the protection against vaginal infections with herpes simplex virus 2 and Chlamydia muridarum. Our data confirm the lack of induction of IFN-ε expression after viral infection and the constitutive expression of IFN-ε by cells of the female but also of the male reproductive organs. Interestingly, when expressed from transfected expression plasmids in 293T, HeLa or Neuro2A cells, the mouse and human IFN-ε precursors were inefficiently processed and secretion of IFN-ε was minimal. Analysis of chimeric constructs produced between IFN-ε and limitin (IFN-ζ) showed that both the signal peptide and the mature moiety of IFN-ε contribute to poor processing of the precursor. Immunofluorescent detection of FLAG-tagged IFN-ε in transfected cells suggested that IFN-ε and chimeric proteins were defective for progression through the secretory pathway. IFN-ε did not, however, act intracellularly and impart an antiviral state to producing cells. Given the constitutive expression of IFN-ε in specialized cells and the poor processing of IFN-ε precursor in fibroblasts and cell lines, we hypothesize that IFN-ε secretion may require a co-factor specifically expressed in cells of the reproductive organs, that might secure the system against aberrant release of this IFN.

Identification of Quiescent, Stem-Like Cells in the Distal Female Reproductive Tract

In fertile women, the endometrium undergoes regular cycles of tissue build-up and regression. It is likely that uterine stem cells are involved in this remarkable turn over. The main goal of our current investigations was to identify slow-cycling (quiescent) endometrial stem cells by means of a pulse-chase approach to selectively earmark, prospectively isolate, and characterize label-retaining cells (LRCs). To this aim, transgenic mice expressing histone2B-GFP (H2B-GFP) in a Tet-inducible fashion were administered doxycycline (pulse) which was thereafter withdrawn from the drinking water (chase). Over time, dividing cells progressively loose GFP signal whereas infrequently dividing cells retain H2B-GFP expression. We evaluated H2B-GFP retaining cells at different chase time points and identified long-term (LT; >12 weeks) LRCs. The LT-LRCs are negative for estrogen receptor-α and express low levels of progesterone receptors. LRCs sorted by FACS are able to form spheroids capable of self-renewal and differentiation. Upon serum stimulation spheroid cells are induced to differentiate and form glandular structures which express markers of mature Műllerian epithelial cells. Overall, the results indicate that quiescent cells located in the distal oviduct have stem-like properties and can differentiate into distinct cell lineages specific of endometrium, proximal and distal oviduct. Future lineage-tracing studies will elucidate the role played by these cells in homeostasis, tissue injury and cancer of the female reproductive tract in the mouse and eventually in man.

Human seminal plasma fosters CD4(+) regulatory T-cell phenotype and transforming growth factor-β1 expression.

Semen mediates expansion of CD4(+) regulatory T cells (Treg) in the murine female reproductive tract. The impact of semen on Treg in humans, however, remains unclear. Using seminal plasma (SP) from 20 healthy donors, we investigated the impact of human semen on CD4(+) T-cell expression of CD127 and CD49d as well as CD4(+) CD127(low) CD49d(low) Treg proliferation, apoptosis, and intracellular expression of FoxP3, TGF-β1, and IL-10. SP reduced CD4(+) T-cell expression of CD127 and CD49d and increased the proportion of CD127(low) CD49d(low) Treg. This increase was non-proliferative, mediated in part via the conversion of CD4(+) helper T cells into FoxP3(-) but not FoxP3(+) Treg. Additionally, SP induced an increase in intracellular expression of the immunosuppressive cytokine TGF-β1 by the FoxP3(-) but not FoxP3(+) Treg. SP had no impact on Treg intracellular expression of IL-10. Human seminal plasma fosters the non-proliferative increase in the proportion and immunoregulatory activity of FoxP3(-) Treg.

Impact of Methotrexate and Leucovorin on Hormonal Regulated Enzymes of Carbohydrate Metabolism in Accessory Reproductive Tissues of Ovariectomized Rats

Background: Female reproductive tract cells are fast proliferating like cancer cells. Methotrexate (MTX), an anticancer drug when used to treat non-neoplastic diseases in young women results in deleterious effects leading to infertility. Oviduct, uterus, cervix and vagina depend on ovary for their structural and functional sustenance. Ovarian ablation followed by chemotherapy in premenopausal women is a common practice in the treatment of cancer. Hence, methotrexate effect on reproductive organs in Ovariectomized (OVX) rats was investigated. Objective: To reveal the direct influence of MTX on estradiol and progesterone in OVX rats. Also, the protective role of leucovorin (LCN) an antidote to MTX, estradiol (E2) and progesterone (P) replacement was investigated. Methods: Animals were randomly divided into the following groups (n=6): group 1: OVX, group 2: OVX+MTX, group 3: OVX+MTX+LCN, group 4: OVX+E2, OVX+MTX+LCN+E2, group 5: OVX+P, group 6: OVX+MTX+LCN+P, treated once/day intramuscularly (im) for 4 days and sacrificed on day 5. At the end of the experiment reproductive 24 tract tissues oviduct, uterus, cervix and vagina were used for total protein, glycogen, Glucose-6-Phosphate Dehydrogenase (G6PD), Lactate Dehydrogenase (LDH) and Alkaline Phosphatase (ALP) enzyme activities. Results: MTX treatment reduced protein, glycogen, G6PD, LDH and ALP activities in oviduct, uterus, cervix and vagina of OVX rats. However, LCN supplementation was partially protective on some enzymes, which was tissue dependent. Hormonal replacement improved protein and enzyme levels in all the tissues. Estradiol treatment showed a distinct enhancement over progesterone in elevating such levels. However, MTX and LCN in a combination treatment with steroids reversed the activities of enzymes in all the tissues.

Nanoparticles Containing siRNA to Silence CD4 and CCR5 Reduce Expression of These Receptors and Inhibit HIV-1 Infection in Human Female Reproductive Tract Tissue Explants

Human Immunodeficiency Virus-type 1 (HIV- 1) binds to CD4 and CCR5 receptors on target cells in the human female reproductive tract. We sought to determine whether reducing levels of messenger RNA (mRNA) transcripts that encode these receptors in female reproductive tract cells could protect mucosal tissue explants from HIV- 1 infection. Explants prepared from the endometrium, endocervix, and ectocervix of hysterectomy tissues from HIV-1 sero-negative women were exposed to nanoparticles containing CD4- and CCR5-specific short-interfering RNA (siRNA) sequences. Explants were then exposed two days later to HIV-1, and HIV-1 reverse transcripts were measured five days post-infection. Explants treated with nanoparticles containing CD4- and CCR5-specific siRNA showed reduced levels of CD4 and CCR5 transcripts, and significantly lower levels of HIV-1 reverse transcripts compared to those treated with an irrelevant siRNA. In female reproductive tract explants and in peripheral blood cell cultures, siRNA transfection induced the secretion of IFN-alpha (IFN-α), a potent antiviral cytokine. In female mice, murine-specific Cd4-siRNA nanoparticles instilled within the uterus significantly reduced murine Cd4 transcripts by day 3. Our findings demonstrate that siRNA nanoparticles reduce expression of HIV-1 infectivity receptors in human female reproductive tract tissues and also inhibit HIV-1 infection. Murine studies demonstrate that nanoparticles can penetrate the reproductive tract tissues in vivo and silence gene expression. The induction of IFN-α after siRNA transfection can potentially contribute to the antiviral effect. These findings support the therapeutic development of nanoparticles to deliver siRNA molecules to silence host cell receptors in the female reproductive tract as a novel microbicide to inhibit mucosal HIV-1 transmission.

Nanoparticles containing siRNA to silence CD4 and CCR5 reduce expression of these receptors and inhibit HIV-1 infection in human female reproductive tract tissue explants

Human Immunodeficiency Virus-type 1 (HIV-1) binds to CD4 and CCR5 receptors on target cells in the human female reproductive tract. We sought to determine whether reducing levels of messenger RNA (mRNA) transcripts that encode these receptors in female reproductive tract cells could protect mucosal tissue explants from HIV-1 infection. Explants prepared from the endometrium, endocervix, and ectocervix of hysterectomy tissues from HIV-1 sero-negative women were exposed to nanoparticles containing CD4- and CCR5-specific short-interfering RNA (siRNA) sequences. Explants were then exposed two days later to HIV-1, and HIV-1 reverse transcripts were measured five days post-infection. Explants treated with nanoparticles containing CD4- and CCR5-specific siRNA showed reduced levels of CD4 and CCR5 transcripts, and significantly lower levels of HIV-1 reverse transcripts compared to those treated with an irrelevant siRNA. In female reproductive tract explants and in peripheral blood cell cultures, siRNA transfection induced the secretion of IFN-alpha (IFN-α), a potent antiviral cytokine. In female mice, murine-specific Cd4-siRNA nanoparticles instilled within the uterus significantly reduced murine Cd4 transcripts by day 3. Our findings demonstrate that siRNA nanoparticles reduce expression of HIV-1 infectivity receptors in human female reproductive tract tissues and also inhibit HIV-1 infection. Murine studies demonstrate that nanoparticles can penetrate the reproductive tract tissues in vivo and silence gene expression. The induction of IFN-α after siRNA transfection can potentially contribute to the antiviral effect. These findings support the therapeutic development of nanoparticles to deliver siRNA molecules to silence host cell receptors in the female reproductive tract as a novel microbicide to inhibit mucosal HIV-1 transmission.

Distribution of eosinophil granulocytes and mast cells in the reproductive tract of female goats in the preimplantation phase

Changes in eosinophil granulocytes and mast cells post-insemination may affect conceptus implantation, but information regarding the numbers of such cells in the mammalian reproductive tract is limited. This study investigated the preimplantation distribution of eosinophil granulocytes and mast cells (MCs) in the reproductive tract organs of female goats. Uterus, uterine cervix and uterine tubes samples were obtained at slaughter. Cornu uteri were washed in phosphate buffer solution (each animal contained at least one embryo). Tissues were fixed in 10% neutral buffered formol, Carnoy solution and Mota's fixative (basic lead acetate) for 48 h and embedded in paraffin. Six-micrometre-thick sections were stained with Congo red (for eosinophil granulocytes) and toluidine blue in 1% aqueous solution at pH 1.0 for 5 min (for MCs). In the uterus, MCs occurred in highest numbers in the myometrium. Higher MC numbers were observed in uterus, uterine and uterine tubes in the preimplantation (experimental) group (cycle synchronised through 7 days intravaginal sponge with 0.3 g P(4)) compared with the control group (P < 0.05). Eosinophil granulocyte numbers were significantly higher in the experimental group than in the control group (P < 0.05). These results indicate preimplantation-related changes in numbers of eosinophil granulocytes and MCs in goat reproductive tract organs.

Distribution and Heterogeneity of Mast Cells in Female Reproductive Tract and Ovary on Different Days of the Oestrus Cycle in Angora Goats

The physiological distribution of mast cells (MCs) in the reproductive tract and ovary of 12 Angora goats was determined using light microscopic histochemical techniques. Uterus (corpus uteri and cornu uteri), uterine cervix, uterine tubes (isthmus and ampulla) and ovary samples were obtained by laparatomy from groups of animals during metoestrus, dioestrus and proestrus (days 5, 10 and 16 of the oestrous cycle). Tissues were fixed in Mota's fixative (basic lead acetate) for 48 h and embedded in paraffin. Six-micrometre-thick sections were stained with toluidine blue in 1% aqueous solution at pH 1.0 for 5 min and alcian blue-Safranin at pH 1.0 for 30 min. MCs were generally associated with blood vessels in all reproductive organs. In the uterus, they were concentrated mainly in the close of the uterine gland and deep stroma in the endometrium. Higher MC numbers were observed by toluidine blue staining in the uterus, uterine cervix and uterine tubes on days 10 (corpus uterine: 4.7 +/- 3.8 and cornu uterine: 4.9 +/- 3.5) and 16 (corpus uterine: 5.9 +/- 4.5 and cornu uterine: 5.4 +/- 2.4) of the oestrous cycle compared with day 5 (p < 0.05). Mast cells were not observed in the follicles, the corpus luteum and the underside of the surface epithelium of the ovarian cortex, but were observed in the interstitial cortical stroma and the ovarian medulla. In the ovary, MC numbers were significantly higher on day 16 of the oestrous cycle (cortex: 3.4 +/- 2.4 and medulla: 5.7 +/- 4.5, p < 0.05). Safranin-positive connective tissue MCs were not observed in the uterine tube on any occasion. These results indicate oestrous cycle-related changes in the number and location of MCs in goat reproductive organs.

Characterization of porcine oviductal epithelial cells, cumulus cells and granulosa cells-conditioned media and their ability to induce acrosome reaction on frozen-thawed bovine spermatozoa

The mammalian female reproductive tract cells have been wildely used in in vitro fertilization. We studied the secretory proteins of porcine oviductal epithelial cells (POEC), and cumulus cells (CC) co-cultured with granulosa cells (GC) in conditioned media (CM), and their ability in inducing acrosome reaction (AR) on frozen-thawed bovine spermatozoa. POEC and CC + GC were cultured in M 199 medium for 48, 96 and 144 h prior to investigation of protein secretion. The results from SDS-PAGE showed secretory proteins sizes of about 17, 22 and >220 kDa in both CM from POEC and CC + GC. To test the ability of CM from POEC and CC + GC in inducing AR, the CM was frozen for 1–3 months before incubating with frozen-thawed bovine spermatozoa. Percentages of the spermatozoa with AR were determined under an inverted microscope and it was found that the tested group 1 (incubated with fresh and frozen CM from POEC for 1–3 months) were 78.44 ± 7.25, 75.78 ± 4.41, 65.22 ± 5.59 and 50.56 ± 6.25, respectively. The tested group 2 (incubated with fresh and frozen CM from CC + GC for 1–3 months) had the percentages of the spermatozoa with AR at 88.67 ± 4.03, 82.22 ± 3.46, 71.00 ± 3.16, and 58.56 ± 4.69, respectively. Statistical analysis of all group percentages indicated that they were significantly different ( P < 0.05). Transmission electron microscope studies demonstrated that there were morphological changes on the sperm heads which resulted in the leakage of acrosin and subsequent AR. We concluded that the CM from both POEC and CC + GC efficiently increased in vitro AR on frozen-thawed bovine spermatozoa. Identification and functions of these secretory proteins are currently under investigation which may lead us to a better understanding of other beneficial effects in the utilization of both CM conditions.

Lymphocytes and MHC class II positive cells in the female rabbit reproductive tract before and after ovulation

In this study, we identified lymphocytes and MHC class II positive (MHC-II+) cells in the reproductive tract of female rabbits both before and after ovulation. CD43+ T cells were frequently present in the mucosa of the oviduct, cervix, and vagina, but far fewer positive cells were seen in the endometrium. The induction of ovulation did not change the cell density in these regions. KEN-5+ T cells and MHC-II+ cells were also frequently seen in the mucosa of the oviduct, cervix, and vagina both before and after ovulation. However, in the uterus, there were very few positive cells before ovulation, but the number increased dramatically after ovulation. Associated with the increase of KEN-5+ T cells, IL-2 mRNA expression in the uterus also increased after ovulation, suggesting that the uterus experienced an increase of T-cell activation. IgM- and IgA-positive B cells were not commonly seen in the reproductive tract and the induction of ovulation did not alter this. Our results suggest that the reproductive tract of female rabbits has the capacity to mount an immune response and that the immune cell distribution of the rabbit reproductive tract has some distinctive features compared with that found in other species.

Leukaemia inhibitory factor in implantation and uterine biology

Leukaemia inhibitory factor (LIF) is one of the most important cytokines in the reproductive tract. Without expression of LIF in the uterus, implantation of a blastocyst cannot begin. Yet, 13 years after publication of the phenotype of the LIF knockout mouse we are only just beginning to understand how LIF functions in the uterus. This review addresses our knowledge of the role of LIF in regulating implantation through its influence on the luminal epithelium and stromal decidualization, but also its influence on reproductive tract cells such as leukocytes and glandular epithelium, during the pre-implantation phase of pregnancy.

Optimizing incubation conditions for the preservation of sperm motility in processed semen samples

To determine the optimal conditions to maintain sperm motility and viability for prolonged periods, semen specimens were processed and cultured in different temperature and conditions. We conclude that processing of semen and incubation of sperm in protein-supplemented N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) human tubal fluid medium at room temperature best preserved viability and motility of sperm.

Immunoexpression of Androgen Receptors in the Reproductive Tract of the Stallion

The objective of this study was to visualize androgen receptors (ARs) in the testis, epididymis, and prostate of the stallion by means of immunohistochemistry. Nuclear immunostaining was found in all somatic cells in the testis--Leydig, Sertoli, and peritubular myoid cells; in both types of epithelial cells of the epididymis; and in the secretory cells of the prostate. These results indicate that ARs are distributed throughout the reproductive tract cells of the stallion.

A method for the dispersal and characterization of leukocytes from the human female reproductive tract.

The isolation of human female reproductive tract (RT) cells that maintain viability and are representative of the entire population is essential for a thorough evaluation of mucosal immunity in the reproductive tract mucosa. Here, we describe the isolation of RT cells in high yields and with high viability from the Fallopian tube, uterine endometrium, endocervix, ectocervix and vagina. This cell dispersion method uses an enzyme cocktail composed of pancreatin, hyaluronidase, and collagenase (PHC), and employs a 250-microm mesh screen to facilitate cell dispersion. The yields of cells isolated per gram of tissue in the presence of this PHC cocktail were compared and found to be strikingly higher relative to the yields obtained with other enzyme cocktails or in the absence of enzymes. Flow cytometry was used to characterize leukocyte subsets isolated from uterine endometrium in the presence of the various enzyme cocktails. The common leukocyte antigen marker CD45, pan T-cell marker CD3, monocyte/macrophage marker CD14 and B-cell marker CD19 were retained after exposure to the PHC cocktail of enzymes. The expression of CD8 and CD4 was lost after exposure to added enzymes but regained after culture overnight. These studies demonstrate the feasibility of using enzymatic digestion for the isolation of whole populations of Fallopian tube, endometrial, cervical and vaginal cells, including leukocyte subsets in high yields, and provide a foundation for investigating mucosal immune cell function in the human female RT.