Mouse Vagina Research Articles

Antigen recognition in the female reproductive tract: I. Uptake of intraluminal protein tracers in the mouse vagina

Local immunization in the vagina of several species elicits immune responses, but little is known about the uptake, processing and recognition of antigens at this site. We investigated the uptake of intravaginally administered tracers using FITC-bovine albumin, FITC-horse ferritin and FITC-horseradish peroxidase in non-pregnant and pregnant mice. Tracers were detected in cells in the vaginal epithelium and stroma at diestrus, proestrus and metestrus, but not at estrus. During pregnancy, racers were present in vaginal cells on Day 6 but not on Day 13. The distribution of tracers in the vagina was the same in all mice. They were present in vaginal epithelium in cells similar to Langerhans' cells and in the stroma in cells that resembled dendritic cells, fibroblasts or macrophages. In some non-pregnant mice, tracers were present in cells adjacent to lymphatic nodules located in the adventitia between the vagina and urethra. Tracers were seen in phagocytic cells lining the marginal and medullary sinuses of the draining lymph nodes (iliac nodes) in some non-pregnant mice at 4 h after intravaginal administration, or in small, dendritic cells in the paracortex at 17 h. To test the possibility that transfer of proteins into the vagina was due to toxic effects of the tracers, FITC-conjugated proteins were also administered into the lumen of uterine horns, and their distribution in horns, cervix and vagina was studied. In uterine horns, tracers were either absent or were located only in apical vesicles in the luminal epithelium. Tracers were present in the cervix and vagina as described above for intravaginal tracers. This result suggests that uptake of tracers in the vagina was not due to toxic effects, and that the vagina and cervix are major sites of protein uptake into the reproductive tract.

Morphological and Biochemical Alterations in Reproductive Tracts of Neonatal Female Mice Treated with the Pesticide Methoxychlor1

Effects of estradiol-17 beta and the estrogenicity of different doses of the technical grade pesticide methoxychlor were compared in the vagina, uterus, and oviducts of neonatal mice. Beginning within 24 h of birth, neonates received 10 daily i.p. injections of sesame oil vehicle, 10.0 micrograms estradiol, or 0.05, 0.1, 0.5, or 1.0 mg methoxychlor. Estradiol injections induced precocious vaginal opening, complete vaginal cornification, and increased total reproductive tract weight and its DNA content. In comparison to the controls, the three highest methoxychlor doses also significantly increased the weights of the reproductive tracts and stimulated their development. The two highest doses (0.5 and 1.0 mg) also induced precocious vaginal opening and complete vaginal cornification. In addition, the same two doses produced atypical cells in the uterus and oviducts that may be indicative of early dysplasia; similar atypia were not recorded following estradiol treatments. Total DNA content in various reproductive organs increased with increased methoxychlor dosages. Dose-response changes were observed in the oviduct and uterus but not vagina. In summary, methoxychlor stimulated the development of neonatal female reproductive tracts, even at concentrations not previously reported to be biologically active. Furthermore, the higher doses induced abnormalities that were not seen following estradiol treatment; these abnormalities may represent precursors of pathological changes.

A recessive mutation causing imperforate vagina in mice.

A recessive mutation (ipv) causing imperforate vagina was discovered in a line of mice selected for low lean tissue mass as a proportion of body weight. Two full sisters were found to have marked swelling of the perineum and complete closure of the vagina. Crosses of heterozygotes identified by progeny testing produced a female progeny ratio not different from the 3 normal: 1 affected (chi 2 = 0.695; p less than .3) expected on the basis of a recessive allele at a single autosomal locus. As a consequence of the imperforate vagina, the uterus and vagina were greatly distended by fluid. The uterus of affected females displayed a swollen uterine lumen and thin endometrial stroma and muscularis. Ovarian tissue of affected females was similar to that of normal mice, and affected females produced ova that were normal in appearance. The mutation causing an imperforate vagina may present a useful model for studying the basis of abnormal vaginal development in other species and increasing the understanding of normal vaginal development in the mouse.

Changes in the uterus and vagina of mice treated neonatally with antiestrogens.

Adenosis-like lesions (ADL) were found in the upper vagina including the cervical region of ICR mice as early as 1 day after 5 daily injections of 200 micrograms tamoxifen (Tx) starting on the day of birth (200-micrograms Tx mice). ADL developed progressively from 5 to 30 days of age. Involution of the musculature and suppression of the gland genesis occurred in the uterus of 5- to 30-day-old, 200-micrograms Tx mice. The uterine abnormality resulted in a loss of type III collagen and laminin, and in an increase in fibronectin and type I collagen in the mesenchymal stroma of 15-day-old, 200-microgram Tx mice. Vaginal ADL and uterine myometrial involution were also encountered in 35-day-old mice given neonatal injections of 2-200 micrograms Tx, 200 micrograms clomiphene (Clm) and 200 micrograms nafoxidine (Naf), respectively. These changes were never observed in control and 0.2-microgram Tx mice. Ovariectomy performed at 10 days resulted in a reduction in the weight of uteri and the extent of ADL occupancy in Tx, Clm and Naf mice. Five daily injections of 0.1 microgram 17 beta-estradiol (E2) beginning at 30 days of age increased uterine weight and ADL occupancy extent in Tx (2-200 micrograms), Clm and Naf mice. The present findings suggest, therefore, that the antiestrogens act as an estrogen agonist to mouse uterus and vagina and that the induced ADL develops progressively depending on ovarian estrogen.

Human Seminal Plasma Suppresses Delayed-Type Hypersensitivity Responses to Intravaginally Deposited Sheep Red Blood Cells and Sperm: Separation of Immunosuppressive Factors

Although significant progress has been made in identifying the immunosuppressive factors in seminal plasma (SP) and their possible action in vitro, the potential role of SP in naturally occurring normal immune responses in vivo is less certain. Human SP or its fractions, administered 5 or 10 times at 3-day intervals into the mouse vagina simultaneously with washed human sperm or sheep red blood cells, suppressed the delayed-type hypersensitivity reactions to footpad injection of the antigens. However, SP failed to suppress antibody formation to these antigens in the applied experimental conditions. In an in vitro study, SP suppressed the proliferation of human peripheral blood mononuclear cells to phytohemagglutinin. This suppression of in vitro proliferation by SP was mediated by materials of both low (LMW) and high molecular weight (HMW). Among the HMW materials, a factor with approximately 1,500 kilodaltons, partially purified by DEAE-cellulose chromatography and gel filtration, exhibited the most potent suppressor activity in vitro and in vivo. The suppressive activities of SP or its fractions were not abolished by treatment with both heat (95 degrees C for 10 min) and trypsin (1.0 mg/ml, 37 degrees C for 4 h). These findings indicate that SP could contribute to the development of suppressed cellular immunity to sperm; immunosuppressive actions of SP are mediated by at least two distinct factors in vivo as well as in vitro.

Estrogen Responsiveness and the Estrogen Receptor during Development of the Murine Female Reproductive Tract: (estrogen receptor/autoradiography/female reproductive tract).

The developing uterus, vagina, and cervix of mice whose age ranged from 16 days of gestation to 90 days postnatal were examined for nuclear estrogen receptors (ERs) by autoradiographic and whole cell uptake techniques. ERs were present within mesenchymal cells of these organs throughout the entire period of development and maturation. By contrast, nuclear ER first became detectable by autoradiography in the epithelium of vagina and uterus at 5 and 6 days postnatal, respectively. As a result of administration of the synthetic estrogen, diethylstilbestrol (DES), consecutively from 16 to 18 days of gestation, uterine and vaginal epithelial cell height was increased and epithelial secretory activity was elevated during the first 48 hr of postnatal life. Also, a single does of DES administered on the 2nd day after birth stimulated epithelial proliferation in the uterus as determined by 3 H-thymidine incorporation. These typical estrogenic effects occurred in the absence of nuclear ER within the epithelium. Prenatal DES treatment accelerated the onset of ER activity within the epithelium by 2 to 3 days relative to controls. The possibility that certain effects of estrogen on epithelial differentiation may be mediated indirectly via ER positive mesenchymal cells is discussed.

Aging and chronic estradiol exposure impair estradiol-induced cornification but not proliferation of vaginal epithelium in C57BL/6J mice.

Long-term exposure of adult female rodents to estrogen has many deleterious effects on reproductive neuro-endocrine structure and function, but its effects on peripheral target tissues are not well known. This study was designed to determine whether chronic exposure of young mice to estradiol (E2) alters the response of the vagina to E2, and if so, whether aging potentiates this alteration. Eight-week-old mice were ovariectomized (ovx) and given subcutaneous Silastic or polyethylene (PE) implants containing E2. Silastic implants produced supra-physiologic E2 levels, while E2 levels in PE-implanted mice were within the physiologic range. Initially all E2-exposed mice showed vaginal cornification (CORN). However, CORN soon began to decline and was virtually absent 3-5 mo after implantation, despite evidence of continued, albeit reduced, release of E2 from the implants. Mice were reimplanted with new E2 implants to determine whether the loss of CORN resulted from an altered response to E2 or from a decreased release of E2 from the implants. Vaginas of mice previously exposed to either Silastic (high E2) or PE (low E2) implants failed to cornify in response to new E2 implants, whereas vaginas of mice that had been initially exposed to implants without E2 cornified in response to identical E2 implants. When old (23 mo) acutely ovx mice were given E2-containing Silastic implants, the peak level and duration of CORN were only one-third and one-fifth, respectively, of that seen in young mice. Non-cornifying epithelia from both young and old chronically E2-exposed mice were as hyperplastic and active mitotically as cornifying epithelia, indicating that the loss of CORN was not a result of decreased epithelial proliferation.(ABSTRACT TRUNCATED AT 250 WORDS)

Enhancement of colonization of Ureaplasma urealyticum in the mouse genital tract by estrogen treatment

Experimental colonization by Ureaplasma urealyticum in the vagina of Balbc mice was greatly enhanced by administration of estrogen, whereas it was not affected by treatment with progesterone. Ureaplasma was recovered from almost half of the estrogen-treated animals 7 days after inoculation, and the organism persisted in all of these animals until 21 days after inoculation; on the other hand, the organism was isolated in less than 20% of the untreated, progesterone-treated, or danazol-treated mice 7 days after inoculation and was no longer isolated in any of these groups 21 days after inoculation. At least a hundredfold more organisms were recovered from the estrogen-treated mice as compared to the untreated mice 14 days after inoculation and thereafter. U. urealyticum was also isolated from the uterine cavity and salpinx in approximately half of the mice in which the organism was isolated from the vagina.

Colonization of congenitally athymic, gnotobiotic mice by Candida albicans

Colony counts, scanning electron microscopy, and light microscopy were used to assess the capacity of Candida albicans to colonize (naturally) and infect the alimentary tract of adult and neonatal (athymic [nu/nu] or heterozygous [+/nu] littermates) germfree BALB/c mice. When exposed to yeast-phase C. albicans, the alimentary tract of adult germfree mice (nu/nu or +/nu) is quickly (within 24 to 48 h) colonized with yeast cells. Neither morbidity nor mortality was evident in any mice that were colonized with a pure culture of C. albicans for 6 months. Yeast cells of C. albicans predominated on mucosal surfaces in the oral cavities and vaginas of adult athymic and heterozygous mice. In both genotypes, C. albicans hyphae were observed in keratinized tissue on the dorsal posterior tongue surface and in the cardial-atrium section of the stomach. Conversely, neonatal athymic or heterozygous mice, born to germfree or C. albicans-colonized mothers, do not become heavily colonized or infected with C. albicans until 11 to 15 days after birth. Although yeast cells adhered to some mucosal surfaces in vivo, neither widespread mucocutaneous candidiasis, i.e., invasion of mucosal surfaces with C. albicans hyphae, nor overwhelming systemic candidiasis was evident in neonatal (nu/nu or +/nu) mice. Thus, even in the absence of functional T-cells and a viable bacterial flora, athymic and heterozygous littermate mice (adult or neonatal BALB/c) that are colonized with a pure culture of C. albicans manifest resistance to extensive mucocutaneous and systemic candidiasis.

Enhancement of experimental Mycoplasma pulmonis infection of the mouse genital tract by progesterone treatment.

Experimental infection of the vagina of TO and CBA mice with Mycoplasma pulmonis was enchanced greatly by progesterone treatment. Almost all treated animals became infected, whereas only two-thirds of the untreated TO mice and less than half of the untreated CBA mice did so. Almost 1000-fold more organisms were recovered from treated than from untreated mice and the duration of infection was more than doubled. The enhanced infection in the hormone-treated animals was accompanied by a more severe vaginal polymorphonuclear leucocyte response which reached a maximum two weeks after inoculation of M. pulmonis. In the TO mice the eventual decline in cellular response coincided with the gradual disappearance of the mycoplasmal infection. The implications of these findings for genital infections of other animal species are discussed.

Immunofluorescence analysis of estrogen receptor in adult and neonatal mouse vagina.

Immunofluorescence analysis of estrogen receptor in adult and neonatal mouse vagina.

The structure and function of oestrogens. VI. The quinone methide hypothesis: stability of the benzylic hydrogen atoms of meso-hexoestrol during oestrogenic stimulation in mice.

A new hypothesis for oestrogen action at the molecular level is that the phenolic moiety of an oestrogen is oxidized to the corresponding quinone methide which is rapidly reduced with, say, NADPH to regenerate the oestrogen; the feedback from the rapid consumption of oxidant and/or reductant in the target organs might be responsible for the local increase in RNA synthesis, and its consequent phenomena. This hypothesis requires continuous removal and replacement of a benzylic hydrogen atom (H9 alpha of oestradiol) in an oestrogen at the receptor site. When a physiological dose of a mixture meso-(2-14C)- and meso-(2,3,4,5-3H4)hexoestrol was administered intravaginally in mice, the 14C:3H ratio in the phenolic portion of the vaginal extracts was essentially the same as that of the pure test mixture. This indicates that there was no net exchange of benzylic hydrogen in meso-hexoestrol during its stimulation of the mouse vagina.

Heterotopic columnar epithelium and adenosis in the vagina of the mouse after neonatal treatment with clomiphene citrate

Physiologically, the epithelium of the mouse vagina undergoes conversion during early postnatal life from columnar to stratified squamous. A similar process in the human occurs in the late first and early second trimesters of pregnancy. The mouse vagina has been identified as a good developmental model of the human. Previous work in the mouse has shown that this process is affected by neonatal administration of diethylstilbestrol. We have administered diethylstilbestrol and clomiphene citrate to parallel groups of BALB/c neonatal mice. They were followed up to 24 weeks. In both groups, persistent columnar or heterotopic columnar epithelium, not seen in the control mice, was identified and associated with adenosis. This effect of diethylstilbestrol and clomiphene citrate appears to be similar to the biologic response to transplacental diethylstilbestrol in the human. After transplacental diethylstilbestrol, malignant vaginal tumors rarely develop. If clomiphene citrate, given to the human prior to pregnancy to induce ovulation or by inadvertence during pregnancy, were to circulate into the critical time of vaginal differentiation, a similar biologic potential may exist. The first situation seems to be unlikely. The second is of more concern.

Autoradiographic analysis of nuclear estrogen binding sites during postnatal development of the genital tract of female mice

Autoradiographic analysis of [ 3H]-estrogen nuclear binding sites was performed on developing genital tracts (uterus, cervix and vagina) of mice 1 to 90 days postpartum. During days 1 to 15 postpartum, nuclear estrogen binding sites were observed exclusively within mesenchymal cells; epithelial cells did not exhibit nuclear labelling. At 18 days postpartum vaginal and cervical epithelial cells exhibited nuclear estrogen binding sites for the first time, whereas the initial appearance of estrogen receptor activity in the epithelium of the uterus was detected at 20 days postpartum. Thereafter, nuclear estrogen binding sites were maintained in both epithelial and stromal cells into adulthood. The acquisition of nuclear binding sites within epithelium of female genital organs at 18 days is discussed in terms of epithelial-mesenchymal interactions and the acquisition of growth responsiveness.

Ultrastructural study of the mucification of the stratified epithelium of the mouse vagina.

After transplacental treatment of mice with estrogens, a heavy mucification was found in the fornices of the vaginae of the offspring. The resulting mucified stratified epithelium is described by light and electron microscopy. It consists of two different cell types: cells forming mucus on top of cells forming tonofilaments. Mucus formation is usually attributed to treatment with progestagens, or with the estrogens combined with vitamin A. Estrogen treatment per se has been shown to be responsible for tonofilament formation. Our unexpected findings are discussed against the background of different theories of development of vaginal epithelium as well as their possible interpretation as beginning of adenosis.

A STUDY ON THE ESTRADIOL-INDUCED AUGMENTATION OF A SPECIFIC CELL PRODUCT IN THE VAGINAL EPITHELIUM OF THE NEONATAL MOUSE.

The epithelial content of an estradiol-sensitive immunological marker (CVA) has been quantified by mixed haemagglutination on tissue sections from the vagina of neonatal mice exposed to different schedules of estradiol treatment. Daily administration of estradiol-17β (5 μg/day) was especially efficient in elevating the CVA content when the hormone was administered during the first four days after birth. Following a single injection of estradiol-17β (5 μg in aqueous suspension) on one of the first five days of life, the vaginal epithelium reacted with a more vigourous CVA accumulation when challenged with estradiol at a later time. The effect was most pronounced for days 2 and 3. The possibility that this early effect of estradiol may involve other mechanisms than those operative in the estradiol action at later stages, is discussed.

Transitional changes in the surface epithelium of the cycling mouse vagina, cervix and uterus: scanning electron microscopic studies.

During the estrous cycle, the mouse reproductive tract undergoes a number of specific morphological changes. The alterations which occur in the luminal surface of the vagina, cervix and uterus were studied by scanning electron microscopy (SEM). The stage of estrus was identified by vaginal observation, cytology and histology. The lumen of the reproductive tract was observed with the SEM at low powers of magnification as well as at the subcellular level. Surface epithelial alterations associated with mucification and keratinization of vaginal epithelium were identified by SEM; changes in the squamocolumnar junction were studied as were the cyclic variations in the mouse endometrium. This study was done to characterize the morphological appearance of the normal female mouse reproductive tract and thereby establish a normal baseline for the evaluation of samples from subsequent studies of corresponding pathologic tissue. A comparison of the similarity of the epithelial characteristics observed in normal cervix and vagina at diestrus to those in cells described by other investigators in reportedly dysplastic tissue demonstrates the need for this type of systematic approach in SEM studies.

Antiviral activity of some beta-diketones. 4. Benzyl diketones. In vitro activity against both RNA and DNA viruses.

The synthesis and in vitro antiviral evaluation of a series of substituted benzyl beta-diketones are described. The introduction of a styryl group onto the phenyl ring enhanced activity against herpesvirus type 2. The 4-methoxystyryl homologue 8 was evaluated extensively in vitro and was found to be effective against both RNA and DNA viruses. Compound 8 was evaluated in the mouse vagina against herpes simplex type 1 and produced a significant increase in survival rate as well as in survival time.

8α-estra-1,3,5(10)-triene-1,3,17β-triol as a special type of estrogen having a high vaginotrophic activity and a low uterotrophic activity in castrated mice

The vaginotrophic and uterotrophic activities of 1-OH-8α-E 2 and its acetate in comparison with E 2 and E 3 were assessed on the basis of the effect on weights, water/fat- and RNA/DNA-contents of mouse vagina and uterus. The multiple doses of 8α-steroids or E 3 sufficient to achieve a maximal effect on the vagina were not enough to stimulate the uterus fully. At various periods of time after multiple injections, 1-OH-8α-E 2 acetate was found to have a higher vaginotrophic potency than the uterotrophic potency, when compared with E 2 as a standard. The study on the time course of organ response to 8α-steroids, E 2 and E 3 after a single injection of daily dose, being equipotent when injected once daily for 3 days, demonstrated that 1-OH-8α-E 2 and E 3 were less vaginotrophic and uterotrophic than E 2 and had a short duration of action. 1-OH-8α-E 2 acetate in comparison with E 2produced a long-lasting vaginotrophic effect and an early regressing uterotrophic effect. It can be confirmed that 8α-steroids, like E 3, are a special type of estrogen having a high vaginotrophic activity and a low uterotrophic activity.

Alkaline phosphatase activity in vagina of mice following neonatal administration of estrogen or androgen.

Alkaline phosphatase activity in vagina of mice following neonatal administration of estrogen or androgen.