Reproductive Performance In Mice Research Articles

Mechanism of endometrial MUC2 in reproductive performance in mice through PI3K/AKT signaling pathway after lipopolysaccharide treatment

The objective of this study was to investigate the effects of exposure to endotoxin on the reproductive performance of humans and animals in pregnancy and delivery period. Mucin is considered to play a critical role in protecting the tissue epithelium. At pregnancy period, the MUC2 expression of uterus in the High LPS group was significantly higher than that in the Control group. The glycosaminoglycans of gland cells were secreted into the uterine cavity to protect the uterus. Then, the MUC2 layer became thinner, and LPS entered the lamina propria of the uterus. The mRNA expression of tight junction proteins showed a marked drop, and morphological damage of the uterus occurred. Subsequently, the glycosaminoglycans of gland cells in the High LPS and Low LPS groups increased with the increasing LPS dose, and the damage to the endometrial epithelium was repaired in female mice at Day 5 postdelivery. A low dose of LPS activated the PI3K/AKT signaling pathways to increase the glycosaminoglycans particles, while a high dose of LPS inhibited the PI3K/AKT signaling pathway to decrease the glycosaminoglycans particles. Taken together, our results suggest that gland cells secreted glycosaminoglycans particles into the uterine cavity by exocytosis to increase the thickness of the mucus layer to protect the uterus and that this process was regulated by PI3K/AKT signaling pathways.

Dietary Fucose Affects Macrophage Polarization and Reproductive Performance in Mice

Intestinal mucus protects epithelial and immune cells from the gut resident microorganisms, and provides growth-promoting factors as mucus-derived O-glycans for beneficial bacteria. A lack of intestinal protective mucus results in changes in the commensal microflora composition, mucosal immune system reprogramming, and inflammation. Previous work has shown that fucose, the terminal glycan chain component of the intestinal glycoprotein Mucin2, and fucoidan polysaccharides have an anti-inflammatory effect in some mouse models of colitis. This study evaluates the effect of fucose on reproductive performance in heterozygous mutant Muc2 female mice. We found that even though Muc2+/− females are physiologically indistinguishable from C57Bl/6 mice, they have a significantly reduced reproductive performance upon dietary fucose supplementation. Metagenomic analysis reveals that the otherwise healthy wild-type siblings of Muc2−/− animals have reduced numbers of some of the intestinal commensal bacterial species, compared to C57BL/6 mice. We propose that the changes in beneficial microflora affect the immune status in Muc2+/− mice, which causes implantation impairment. In accordance with this hypothesis, we find that macrophage polarization during pregnancy is impaired in Muc2+/− females upon addition of fucose. Metabolic profiling of peritoneal macrophages from Muc2+/− females reveals their predisposition towards anaerobic glycolysis in favor of oxidative phosphorylation, compared to C57BL/6-derived cells. In vitro experiments on phagocytosis activity and mitochondrial respiration suggest that fucose affects oxidative phosphorylation in a genotype-specific manner, which might interfere with implantation depending on the initial status of macrophages. This hypothesis is further confirmed in BALB/c female mice, where fucose caused pregnancy loss and opposed implantation-associated M2 macrophage polarization. Taken together, these data suggest that intestinal microflora affects host immunity and pregnancy outcome. At the same time, dietary fucose might act as a differential regulator of macrophage polarization during implantation, depending on the immune status of the host.

Melatonergic systems of AANAT, melatonin, and its receptor MT2 in the corpus luteum are essential for reproductive success in mammals†

Corpus luteum (CL) plays a critical role in mammalian reproductive physiology. Its dysfunction will lead to infertility or habitual abortion. In the current study, by use of melatonin specific membrane receptor 2 (MT2) knocking out (KO) mice model combined with RNA-Seq, immunohistochemistry, and immunofluorescence analyses, the genes of melatonin synthetic enzyme arylalkylamine N-acetyltransferase (AANAT) and MT2 were identified to strongly express in the CL of sows and mice. KO MT2 significantly impaired the reproductive performance in mice indicated by the reduced litter sizes. Melatonin treatment elevated the progesterone production in sows suggesting the improved CL function. Mechanistic analysis showed that melatonin upregulated a set of progesterone synthesis-related genes including cytochrome P450 family 11 subfamily A member 1 (Cyp11a1), aldo-keto reductase family 1, member C18 (Akr1c18), isopentenyl-diphosphate delta isomerase 1 (Idi1), and luteinizing hormone/choriogonadotropin receptor (Lhcgr). The upregulation of these genes directly related to the increased progesterone production. The regulatory effects of melatonin on these gene expressions were mediated by MT2 and MT2KO diminished the effects of melatonin in this respect. Thus, the presence of melatonergic system of AANAT, melatonin, and its receptor MT2 in CL is essential for reproductive success in mammals.

Matrix metalloproteinases improves trophoblast invasion and pregnancy potential in mice

Successful implantation is closely linked to the expression of MMP-2 and MMP-9, which greatly influence the ability of an embryo to degrade the basement membrane of the uterine epithelium, mainly composed of type IV collagen, and invade the uterine stroma. The objective of this study was to determine the effect of MMP-2 and MMP-9 co-transfer with embryos on reproductive performance in mice. Using invasion assay, we tested the effect of MMP-2 and MMP-9 for their ability to support trophoblastic invasion in vitro. We performed co-transfer of MMP-2 and MMP-9 with mouse embryos to 2.5 days post-coitum (dpc) pseudo-pregnant uteri using nonsurgical embryo transfer (NSET) technique and evaluated the pregnancy outcomes. Uterine tissue samples were collected to determine collagen content by Masson’s trichrome staining. Our results showed that in vitro treatment of MMP-2 and MMP-9 significantly promoted both spreading and invasion of mouse trophoblastic cells compared to the non-treated blastocysts. Moreover, embryo transfer results showed that MMP-9 co-transfer enhanced pregnancy outcome inform of live pup rate by degrading the extracellular matrix, collagen, and facilitate embryo implantation. Taken together our findings imply that MMP-9 can regulate trophoblastic cell invasion during preimplantation, which may have important consequences on embryo implantation, and shed the light on new strategies to avoid miscarriage and provides a platform for successful human embryo transfer technologies.

The beneficial effects of Moringa oleifera leaf on reproductive performance in mice.

Moringa oleifera is a tropical plant with high nutritional and medicinal value. Recent studies have reported its remarkable effects in inflammatory, antioxidative, and anti‐diabetes modulations, but there was no significant report on its role in animal breeding. In this study, we investigated the effects of dietary Moringa oleifera leaf (MOL) on reproductive performances in mice. We studied the reproductive performance of mice for six consecutive gestations. Mice fed with 4% MOL diet showed improved litter size, litter birth weight, and litter survivals until weaning age compared to control mice fed with normal diet (p < 0.05). Mice fed with MOL diet did not change weight and organ coefficients. Serum malondialdehyde (MDA) concentrations in both male and female mice were significantly decreased by dietary MOL (p < 0.05), but glutathione peroxidase (GSH‐PX) and superoxide dismutase (SOD) were unchanged. For male, dietary MOL significantly reduced sperm abnormality rate (p < 0.05) and Bcl2‐associated X protein (Bax) expression in testis (p < 0.05), but did not affect serum testosterone and the expression levels of androgen receptor (AR), phosphoglycerate kinase 2 (Pgk2), protamine2 (Prm2), and B cell leukemia/lymphoma 2 (Bcl2) in testis. For female, dietary MOL did not change serum estradiol and the expressions of estrogen receptor beta (ERβ), Bcl2, Bax, and vascular endothelial growth factor receptor (VEGFR) in ovary. In summary, MOL increased litter size and antioxidant ability, reduced the rate of sperm abnormality and the expression of Bax. Therefore, MOL may serve as a functional feed addictive for improving animal reproductive performance.

The role of propofol on mouse oocyte meiotic maturation and early embryo development

SummaryPropofol is a intravenous anaesthetic most commonly used in ultrasound oocyte retrieval. We studied if the use of propofol had an effect on mouse oocyte maturation, pregnancy, childbirth and progeny and investigated the correlation between propofol side effects and reproductive performance in mice. There was no statistical difference in mating, pregnancy, childbirth, litter size, the number of stillbirths and survival between each group (P>0.05). Propofol also had no effect on polar body extrusion in oocyte maturation as well as on pronucleus formation and, subsequently, early embryo development (P>0.05). An increased concentration of propofol had no effect on this result, although propofol at more than 0.01 mg/ml reduced polar body extrusion. Different concentrations of propofol had no effect on oocyte culture in vitro, pronucleus formation and early embryo development.

Superovulation at a specific stage of the estrous cycle determines the reproductive performance in mice

Inconsistent reproductive performance has been reported in superovulated mice. Hence, the aim of this study was to analyze the effect and possible mechanism of superovulation timing on mouse reproductive performance. The results showed that mice superovulated at the metestrous (23.08±6.08%) and diestrous stages (33.33±11.45%) presented significantly lower pregnancy rates compared with those superovulated at the estrous stage (66.67±9.20%). After superovulation at the proestrous and estrous stages, mucin 1 (MUC1) and let-7a/let-7b microRNA (miRNA) expression levels were significantly attenuated and enhanced on embryonic day 3.5 (E3.5), respectively, whereas no significant differences in the expression level were found in mice superovulated at the other two stages. A higher number of developing and Graafian follicles was observed in the ovarian sections 48h after the administration of pregnant mare serum gonadotropin (PMSG) at the proestrous and estrous stages. The sections from mice treated at the metestrous and diestrous stages, however, presented more corpora lutea. Therefore, mice superovulated at the proestrous and estrous stages exhibited the best pregnancy rates. Furthermore, the disordered expression of MUC1 and let-7a/let-7b miRNA in mice superovulated at the metestrous and diestrous stages may impair reproduction performance.

The Clock mutation reduces reproductive performance of mice by affecting the implantation capacity: Maternal Clock mutation is not the only factor affecting implantation

Here, we showed that the Clock gene was important for reproductive performance in mice. We compared outcomes from the four possible mating combinations between wild-type mice (WT) and mice homozygous for the Clock delta-19 mutation (CL). We found that the only significant differences were between the WT♂ × WT♀ and CL♂ × CL♀ mating groups; these groups differed with regard to elongation of the pregnancy period (19.3 vs. 20.5 days, respectively, P < 0.05) and the number of newborn pups (13.4 ± 0.8 vs. 8.6 ± 1.5, respectively, P < 0.05). Because CL dams impregnated by male CLs exhibited normal continuous increases in body weight during the entire gestation period and did not show any signs of spontaneous abortion from mid to late gestation, we reasoned that some embryos were lost before or at the time of implantation. Immediately before implantation (88 hours after fertilization), neither the number of embryos collected from uteri nor the percentage of the embryos that reached the blastocyst stage differed significantly among mating groups. In contrast, immediately after implantation (160 hours after fertilization), the average number of implantation sites was significantly lower for the CL♂ × CL♀ mating group than that for the WT♂ × WT♀ mating group (7.0 vs. 13.0, P < 0.05); this decrease was accompanied by a significant lowering of the positions of implantation sites in uteri, and this lowering of the implantation sites was more severe when mothers and embryos bore more CL alleles (WT♂ × WT♀ >CL♂ × WT♀ > WT♂ × CL♀ >CL♂ × CL♀), suggesting that the Clock mutation reduced the reproduction performance of the parents by affecting the implantation capacity via such as embryos' ability to implant.

Limits to sustained energy intake. XIV. Heritability of reproductive performance in mice

Limits to sustained energy intake (SusEI) are important because they constrain many aspects of animal performance. Individual variability in SusEI may be imposed by genetic factors that are inherited from parents to offspring. Here, we investigated heritability of reproductive performance in MF1 mice. Food intake, milk energy output (MEO) and litter mass were measured in mothers (F0) and daughters (F1) that were raising litters of 10 pups. Cross-fostering was designed so that half of each litter consisted of biological offspring and the rest came from one unrelated female (i.e. fostered pups). Food intake increased linearly during early lactation and reached a plateau during late lactation (day 9-13, called the asymptotic food intake, FIAS, equivalent to SusEI). Parent-offspring regression showed that FIAS, MEO and litter mass were all positively and significantly related between mothers and their biological daughters, but no significant relationships were found between the same traits for mothers and fostered daughters. FIAS at peak lactation was significantly correlated to adult food intake and body mass when the mice were 6 months old and not lactating. In conclusion, a large part of the variation in FIAS could be explained by genetic variation or maternal effects in pregnancy whereas non-genetic maternal effects in lactation were negligible. As a consequence, biological daughters of mothers with high reproductive performance (i.e. high milk production and hence higher litter mass at weaning) had a better reproductive performance themselves, independent of the mother that raised them during lactation.

Reproductive toxicity of gamma-ethyl-gamma-phenyl-butyrolactone, a new anticonvulsant and hypnotic drug, in mice. Fertility and peri- and post-natal studies.

The reproductive toxicity of gamma-ethyl-gamma-phenyl-butyrolactone (EPBL, CAS 53380-21-5), a new anticonvulsant and hypnotic drug, was investigated by performing fertility and peri- and post-natal studies in mice. In both studies, EPBL was administered by gavage at 0, 50, 10, or 200 mg/kg doses. The first study found parent toxicity as measured by reduced body weight at the higher dose. At the same dose, the number of live fetuses was decreased. However, neither male or female fertility nor the reproductive performance of mice were affected. The results of the second study showed that the only negative effects were a depression of maternal weight gain and a decrease in body weight of the litter at birth which was 200 mg/kg. A reduction in the survival rate was also seen. There was no evidence that treatment of F0 females from day 15 of pregnancy to day 21 post-partum affected either the reproductive capacity of the F1 offspring or the development of F2 progenies. The non observed effects levels (NOEL) for both studies can be estimated at 100 mg/kg.

Effect of dietary arginine supplementation on reproductive performance of mice with porcine circovirus type 2 infection.

The objective of this study was to investigate whether supplemental dietary arginine increases reproductive performance in mice infected with porcine circovirus type2 (PCV2). A total of 50KM female mice were allotted randomly to the arginine group (0.6% arginine + gestation diet) and control group (1.22% alanine + gestation diet). All the mice began to mate after 14 days of treatment with our prepared feed and challenged with PCV2 at the dose of 100 TCID50 (50% tissue culture infection dose, TCID50) after 7 days of pregnancy. Abortion rate, litter number, litter birth weight, the daily weight gain in the first 7 days and survival rate in the first 2 weeks of the neonates were calculated. The serum progesterone, estrogen, nitric oxide and superoxide dismutase (SOD) activity and total antioxidant capacity (T-AOC) on the 14th day of pregnancy were measured. Arginine supplementation decreased the abortion rate of pregnant mice and mortality of neonates caused by PCV2 infection. Further, litter number, litter birth weight and the daily weight gain of neonates increased in the arginine group compared to the control group. Arginine supplementation increased significantly the serum progesterone (P < 0.01) and nitric oxide levels (P < 0.05), but had little effect on the serum estrogen level. SOD activity and T-AOC in the arginine group were significantly higher (P < 0.01) than the control group. In conclusion, arginine supplementation partially reversed the reproductive failure in mice caused by PCV2 infection.

Effect of vitamin E on reproductive function in the mice treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin*

The study was conducted to determine the effects of vitamin E on reproductive performance in mice treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The 75 female and 40 male mice were randomly assigned into five groups. The levels of both TCDD and vitamin E given by gavage were 0 and 0 (Control group), 100 and 0 (experimental group I), 100 and 20 (experimental group II), 100 and 100 (experimental group III), and 100 ng/kg/day and 500 mg/kg/day (experimental group IV), respectively. Males and females were mixed to mate at the ratio of 1:2 after 4-week experiment. The gavage treatments were continued until the end of gestation in female mice after mating. The results showed that the litter number, survival rate, and body weight at birth of offspring in experimental group I had significantly decreased, and the females' pregnancy rate and pup sex ratio in experimental group I had the decreasing tendencies when compared with the control group. The litter number in experimental group III, survival rate in experimental group II and III, body weight at birth in experimental group III and IV exhibited significant increase compared with experimental group I. The female pregnancy rate in both experimental group III and IV recovered to 100%, but there were no significant differences when compared with experimental group I. The pups' sex ratio had a gradually increasing tendency with increase of vitamin E level, but there was no significant difference among experimental group I-IV. The results suggest that TCDD could induce reproductive toxicity in mice, whereas vitamin E alleviated adverse effects on reproductive performance in mice caused by TCDD.

Effects of Isomeric Fatty Acids on Reproductive Parameters in Mice

The aim of this study was to determine if dietary fatty acids (FA) level or isomeric FA type may affect reproductive parameters in mice. of study Mice were fed for 1 month diets differing in cisFA (cFA) content or type of isomeric FA. Resorption, number of fetuses and placental cytokine expression were determined and sperm acrosome reaction was evaluated after induction by calcium ionophore. Mice fed high fat diets showed increased fetal resorptions, a decrease in interleukin (IL)-4 placental expression in the first generation and an increase of tumor necrosis factor-alpha (TNF-alpha) in the second generation. In this generation, conjugated linoleic acid (CLA) returned TNF-alpha to normal levels and diminished IL-4 and transforming growth factor-beta (TGF-beta) expressions; males fed transFA (tFA) and CLA showed a lower rate of induced acrosome reaction. The amount and type of dietary FA may affect reproductive performance in mice by affecting sperm membrane functionality and placental cytokine production.

Identification of Genetic Regions of Importance for Reproductive Performance in Female Mice

Both environmental and genetic factors can dramatically affect reproductive performance in mice. In this study we have focused on the identification of genetic regions, quantitative trait loci (QTL), which affect the breeding capacity of female mice. We have identified polymorphic microsatellite markers for the mouse strains used and performed a genomewide scan on 237 females from a gene-segregating backcross between a high breeder and a relatively poor breeder. The high-breeder mouse strain we used is the inbred NFR/N mouse (MHC haplotype H-2q), which has extraordinary good breeding properties. The moderate breeder chosen for F(1) and N2 progeny was B10.Q, which is a genetically well-characterized MHC-congenic mouse of the H-2q haplotype. Each of the 237 females of the N2 generation was allowed to mate twice with MHC-congenic B10.RIII (H-2r) males and twice with B10.Q males. A predetermined number of phenotypes related to reproductive performance were recorded, and these included litter size, neonatal growth, and pregnancy rate. Loci controlling litter size were detected on chromosomes 1 (Fecq3) and 9 (Fecq4). The neonatal growth phenotype was affected by Fecq3 and a locus on chromosome 9 (Neogq1). On chromosome 11 two loci affecting the pregnancy rate (Pregq1 and Pregq2) were identified. Furthermore, on chromosomes 13 and 17 we found loci (Pregq3 and Pregq4) influencing the outcome of allogeneic pregnancy (allogeneic by means of MHC disparity between mother and fetuses). A locus on chromosome 1 affecting maternal body weight was also identified and has been denoted Bwq7. It is well known that reproductive performance is polygenically controlled, and the identification of the major loci in this complex process opens the possibility of investigating the natural genetic control of reproduction.

Effects of inorganic mercury on reproductive performance of mice

Effects of mercuric chloride (MC) on the reproductive performance of mice were evaluated. Both male and female mice were divided into four groups that were subsequently exposed to 0.00, 0.25, 0.50, and 1.00 mg/kg/day of MC, respectively. At the end of pre-mating dosing, males were paired with females receiving the same dose. Dosing continued for males throughout mating, while dosing in females continued throughout mating, gestation, and lactation. The males were necropsied at the conclusion of mating and the females were necropsied at the conclusion of lactation. Fertility indices, parturition, gestation, live birth litter size, survival indices, and implantation efficiency were recorded. Subsequently, these data were statistically analyzed. Fertility and survival indices were significantly reduced in the treated groups. Exposure of mice to MC did not affect their litter size. No evidence of mercury induced target organ toxicity was seen in either the clinical pathology parameters or histomorphologic evaluations. However, in MC treated females, ovary weights were significantly different from the control. There were no histomorphologic or clinical pathology effects induced by MC. These results suggested that oral exposure to 0.25–1.00 mg/kg/day of MC produced adverse effects on the reproductive performance of mice in the absence of overt mercury toxicity.

A 13-week subchronic intravaginal toxicity study of the novel broad-spectrum anti-HIV and spermicidal agent, N-[2-(1-cyclohexenyl)ethyl]-N'-[2-(5-bromopyridyl)]-thiourea (PHI-346) in mice.

The nonnucleoside inhibitor (NNI) of HIV-1 reverse transcriptase, PHI-346 (N-[2-(1-cyclohexenyl)ethyl]-N'-[2-(5-bromopyridyl)]-thiourea), is a dual-function spermicidal agent with potent anti-HIV activity against drug-sensitive as well as drug-resistant HIV-1 strains with genotypic and phenotypic NNI resistance. PHI-346 was formulated via a lipophilic gel-microemulsion for intravaginal use as a potential dual-function microbicide. To evaluate the toxicity potential of short-term intravaginal exposure to PHI-346, groups of 15 female B6C3F1 and CD-1 mice were exposed intravaginally to a gel-microemulsion containing 0, 0.5, 1.0, or 2.0% PHI-346, 5 days per week for 13 consecutive weeks. On a molar basis, these concentrations of PHI-346 are 350 to 1,400-times higher than its spermicidal EC50 and nearly 5 x 10(6) to 2 x 10(7) times higher than its in vitro anti-HIV IC50. After 13 weeks of intravaginal treatment, B6C3F1 mice were evaluated for survival, body weight gain, absolute and relative organ weights. Blood was analyzed for hematology and clinical chemistry profiles. Microscopic examination was performed on hematoxylin and eosin-stained tissue sections from each study animal. Placebo control and PHI-346 dosed female CD-1 mice were mated with untreated males in order to evaluate if PHI-346 has any deleterious effects on the reproductive performance. There were no treatment-related mortalities. Mean body weight gain during the dosing period was not reduced by PHI-346 treatment. The hemogram or blood chemistry profiles revealed lack of systemic toxicity following daily intravaginal instillation of PHI-346 for 13 weeks. No clinically significant changes in absolute and relative organ weights were noted in PHI-346 dose groups. Extensive histopathological examination of tissues revealed no treatment-related abnormalities in any of the three PHI-346 dose groups. Repeated intravaginal exposure of CD-1 mice to increasing concentrations of PHI-346 for 13 weeks had no adverse effect on their subsequent reproductive capability, perinatal outcome, growth, and development of offspring. Collectively, these findings demonstrate that repetitive intravaginal administration of PHI-346 at concentrations as high as 1,400-times its spermicidal EC50 and 2 x 10(7) times its in vitro anti-HIV IC50 was not associated with local or systemic toxicity and did not adversely affect the reproductive performance in mice. PHI-346 may be useful as an active ingredient of a safe vaginal microbicide for prevention of the sexual transmission of multidrug-resistant HIV-1.

Endocrine alterations and signaling changes associated with declining ovarian function and advanced biological aging in follicle-stimulating hormone receptor haploinsufficient mice.

Reproductive aging in female mammals is characterized by a progressive decline in fertility due to loss of follicles and reduced ovarian steroidogenesis. In this study we examined some of the endocrine and signaling parameters that might contribute to a decrease in ovulation and reproductive performance of mice with haploinsufficiency of the FSH receptor (FSH-R). For this purpose we compared ovarian changes and hormone levels in FSH-R heterozygous (+/-) and wild-type mice of different ages (3, 7, and 12 mo). Hormone-induced ovulations in immature and 3-mo-old +/- mice were consistently lower. The number of corpora lutea (CL) were lower at 3 and 7 mo, and none were present in 1-yr-old +/- females. The plasma steroid and gonadotropin levels exhibited changes associated with typical ovarian aging. Plasma FSH and LH levels were higher in 7-mo-old +/- mice, but FSH levels continued to rise in both genotypes by 1 yr. Serum estradiol and progesterone were lower in +/- mice at all ages, and testosterone was several-fold higher in 7-mo-old and 1-yr-old +/- mice. Inhibin alpha (Western blot) appeared to be lower in +/- ovaries at all ages. FSH-R (FSH* binding) declined steadily from 3 mo and reaching the lowest point at 1 yr. LH receptor (LH* binding) was high in the 1-yr-old ovary, and expression was localized in the stroma and interstitial cells. Our findings demonstrate that haploinsufficiency of the FSH-R gene could cause premature exhaustion of the gonadal reserves previously noted in these mice. This is accompanied by age-related changes in the hypothalamic-pituitary axis. As these features in our FSH-R +/- mice resemble reproductive failure occurring in middle-age women, further studies in this model might provide useful insights into the mechanisms underlying ovarian aging.

Subchronic (13-week) toxicity studies of intravaginal administration of spermicidal vanadocene acetylacetonato monotriflate in mice

Bis-cyclopentadienyl complexes of vanadium(IV) or vanadocenes are rapid and potent inhibitors of human sperm motility with potential as a new class of contraceptive agents. In this study, groups of 10 B 6C 3F 1 and 20 CD-1 female mice were exposed intravaginally to a gel-microemulsion containing 0, 0.06, 0.12, or 0.25% of a representative vanadocene, vanadocene acetylacetonato monotriflate (VDACAC), five days per week for 13 consecutive weeks. The doses of VDACAC used were nearly 300- to 1250-fold higher than its in vitro spermicidal EC 50 value. After 13 weeks of intravaginal treatment, B 6C 3F 1 mice were evaluated for survival, body weight gain, absolute and relative organ weights, and systemic toxicity. Blood was analyzed for hematological and clinical chemistry profiles. Microscopic examination was performed on hematoxylin- and eosin-stained tissue sections from each study animal. Vanadium content in tissues was determined by atomic absorption spectroscopy. Gel-microemulsion (placebo) control and VDACAC dosed female CD-1 mice were mated with untreated males in order to evaluate if VDACAC has any adverse effects on the reproductive outcome. There were no treatment-related mortalities in either study. Mean body weight gain during the dosing period was not reduced by VDACAC treatment. Hemograms or clinical chemistry profiles did not reveal any toxicologically significant changes attributed to VDACAC treatment. No clinically significant dose-dependent changes in absolute and relative organ weights were noted in VDACAC dose groups. Extensive histopathological examination of tissues revealed no treatment-related abnormalities in any of the three VDACAC dose groups. Vanadium was not incorporated in mouse tissues at levels above 1 μg/g. Repeated intravaginal exposure of CD-1 mice to increasing concentrations of VDACAC for 13 weeks had no adverse effect on their subsequent reproductive capability (100% fertile), neonatal survival (>96%) or pup development. Collectively, these findings demonstrate that repetitive intravaginal administration of VDACAC to yield effective spermicidal concentrations (<0.1%) in the vagina was not associated with systemic toxicity and did not adversely affect the reproductive performance in mice. The spermicidal vanadocene-chelated complex, VDACAC, may be useful as a safe vaginal contraceptive.

Subchronic (13-week) toxicity studies of intravaginal administration of spermicidal vanadocene dithiocarbamate in mice

Spermicidal organometallic complexes of vanadium(IV) with bis(cyclopentadienyl) rings or vanadocenes are a new class of experimental contraceptive agents. In a systematic search for vanadocenes with selective spermicidal activity, we identified vanadocene dithiocarbamate (VDDTC) as the most potent and stable spermicidal compound. In this study, groups of 10 B 6C 3F 1 and 20 female CD-1 mice were exposed intravaginally to a gel-microemulsion containing 0, 0.06, 0.12, and 0.25% VDDTC 5 days per week for 13 consecutive weeks. The doses of VDDTC used were nearly 1250- to 5000-fold higher than its in vitro spermicidal EC 50 value. After 13 weeks of intravaginal treatment, B 6C 3F 1 mice were evaluated for survival, body weight gain, absolute and relative organ weights, and systemic toxicity. Blood was analyzed for hematologic and clinical chemistry parameters. Microscopic examination was performed on hematoxylin and eosin-stained tissue sections from each study animal. Vanadium content in tissues was determined by atomic absorption spectroscopy. Placebo control and VDDTC-dosed female CD-1 mice were mated with untreated males to evaluate whether VDDTC has any deleterious effects on the reproductive performance. There were no treatment-related effects on survival and mean body weight and mean body weight gain during the dosing period. The blood chemistry or hemogram profiles did not reveal any toxicologically significant changes that could be attributed to VDDTC treatment. No clinically significant changes in absolute and relative organ weights were noted in VDDTC dose groups. Extensive histopathological examination of tissues revealed no treatment-related abnormalities in any of the three VDDTC dose groups. The vanadium content of all mouse tissue analyzed was <1 μg/g. Repeated intravaginal exposure of CD-1 mice to increasing concentrations of VDDTC for 13 weeks had no adverse effect on their subsequent reproductive capability (100% fertile), neonatal survival (>90%), or pup development. Collectively, these findings demonstrate that repetitive intravaginal administration of VDDTC to yield effective spermicidal concentrations (<0.1%) in the vagina was not associated with systemic toxicity and did not adversely affect the reproductive performance in mice. VDDTC may have clinical utility as an active ingredient of non-detergent type, safe, vaginal spermicidal contraceptives.

Reproductive performance in mice selected for divergence in body fat content

Correlated responses in female reproductive performance were studied in lines of mice selected for high (HF) and low (LF) fat content relative to a randomly selected control line (RC). Selection was based on an indicator trait of body fat percentage in males (epididymal fat pad weight as a percentage of body weight at 12 weeks of age). LF mice showed a positive correlated response (P < 0.05) in number of pups born (NB), whereas HF showed no significant change. The increase in NB could be explained partially by an increase in ovulation rate (adjusted for body weight at mating; P < 0.01), measured as number of corpora lutea (CL) 16 days postcoitus (P < 0.05) or number of ova (OR) shed 2 days postcoitus (P < 0.01). Asymmetry was observed for CL (P < 0.05) and OR (P < 0.01); the major correlated response was observed in the LF line. Preimplantation and total embryo survival were higher (P < 0.01) in LF than in HF mice, whereas no significant divergence was found for post-implantation embryo survival. Reciprocal crosses of HF and LF lines allowed partitioning of divergence in OR into maternal (P < 0.10) and individual (P < 0.10) effects with LF being larger than HF. No significant heterosis was found for OR or mating weight. Body weight at mating was larger in HF females than in RC controls, while LF females were larger than controls in one experiment and not different from controls in a second experiment. Therefore, body weight at mating did not explain the positive correlated response in ovulation rate observed in the LF line. ZUSAMMENFASSUNG: Reproduktion in gegensätzlich auf Körperfett selektierten Mäusen Korrelierte Selektionswirkungen auf weibliche Reproduktion wurden in Linien untersucht, die auf hohen (HF) oder niedrigen (LF) Fettgehalt selektiert worden waren, relativ zu einer zufallsgepaarten Kontrolle. Selektion beruhte auf Nebenhodenfett, korrigiert auf 12 Wochengewicht, als Indikator-merkmal. LF Mäuse hatten signifikant höhere Zahl geborener Junge (NB), HF zeigten keine signifikanten Wirkungen. Die Steigerung von NB kann teilweise durch hochsignifikante Steigerung der gewichtsadjustierten Ovulationsrate, gemessen als Zahl von Gelbkörper (CL) 16 Tage postcoitus oder Zahl Eier (OR) 2 Tage pc, erklärt werden. Asymmetrie zeigte sich für CL (p < .01) und OR (p < .1), aber keine signifikanten Unterschiede zeigten sich für Post-Implantations Überleben. Reziproke Kreuzungen von HF und LF erlaubten Aufteilung der OR Unterschiede in maternal (p < .1) una individuell verursachte (p < .1) Wirkungen, wobei LF größer als HF waren, aber sich keine Heterosis für OR oder Gewicht bei der Paarung ergab. Dieses war in HF Tieren höher als in RC Kontrollen, was bei LF Tieren in einem, aber nicht im zweiten Versuch zutraf, so daß Körpergewicht nicht die korrelierte Wirkung auf OR erklären kann.