F1 Generation Rats Research Articles

Fisetin impedes developmental methylmercury neurotoxicity via downregulating apoptotic signalling pathway and upregulating Rho GTPase signalling pathway in hippocampus of F1 generation rats

Methyl mercury is a teratogenic and neurodevelopmental toxicant in the environment. MeHg affects several biological pathways critical for brain development. The present study validated the effect of Fisetin on developmental MeHg exposure induced alterations in mitochondrial apoptotic pathway and Rho GTPase mRNA expressions in hippocampus of F1 generation rats. Pregnant Wistar rats were grouped as Group I : administered with vehicle control, Group II: MeHg (1.5 mg/kg b.w), Group III: MeHg + Fisetin (10 mg/kg b.w), Group IV: MeHg + Fisetin (30 mg/kg b.w), Group V: MeHg + Fisetin (50 mg/kg b.w), Group VI: MeHg + Fisetin (70 mg/kg b.w), Group VII: Fisetin (30 mg/kg b.w) alone. Fisetin reduced mercury accumulation in offspring brain. In hippocampus, Fisetin preserved mitochondrial total thiol status, glutathione antioxidant system, mitochondrial metabolic integrity and respiratory chain activity. Fisetin ameliorated apoptotic signals by preventing Cytochrome c release, down regulating ERK 1/2 and Caspase 3 gene expression. Fisetin also upregulated mRNA expressions of RhoA/Rac1/Cdc42 in hippocampus. Predominant effect of Fisetin was to reduce mercury accumulation in offspring brain there by diminishing the toxic effect of MeHg. Hence we showed that, gestational intake of Fisetin (30 mg/kg b.w.) impedes developmental MeHg neurotoxicity by regulating mitochondrial apoptotic and Rho GTPase signalling molecules and by reducing the mercury accumulation in hippocampus of F1 generation rats

Perinatal exposure to 4-nonylphenol can affect fatty acid synthesis in the livers of F1 and F2 generation rats.

Objective: To explore the effects of different dosages of 4-nonylphenol (4-NP) on the fatty acid synthesis and estrogen receptor α (ERα) expression in the livers of F1 and F2 rats. Method: Pregnant rats were randomly divided into four groups: control, NP-5 (5 μg per kg per day), NP-25 (25 μg per kg per day) and NP-125 (125 μg per kg per day). 4-NP was gavaged from gestation day (GD) 6 to postnatal day (PND) 21. Some female rats from the experimental groups were mated with male rats from the control group to obtain the F2 rats. F1 generation rats (23 weeks old) and F2 generation rats (13 weeks old) were killed to detect blood biochemistry and the expression of genes and proteins. Results: Compared with the control group, 4-NP (NP-5, NP-25 and NP-125) can increase the liver organ coefficient of the F1 male offspring (P < 0.05 or P < 0.01). The concentration of high density lipoprotein (HDL) in the F1 female NP-5 group was significantly higher than that of the control group (P < 0.01); other indicators had not changed, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC) and low density lipoprotein (LDL). As the dosage of 4-NP increased, more significant changes of blood biochemistry were found, especially in the NP-125 rats (P < 0.05 or P < 0.01). The changes of histopathology by liver biopsy were consistent with biochemical indices of blood (P < 0.05 or P < 0.01). Compared with the control group, the expression of genes involved in fatty acid synthesis increased significantly (P < 0.05 or P < 0.01), and the degrees of increase were proportional to the dose of 4-NP, as measured by lipoprotein lipase (Lpl), fatty acid synthetase (Fas), sterol regulatory element-binding protein 1 (Srebp-1) and peroxisome proliferator-activated receptor (Ppar)-γ. The expression of genes and proteins of ERα were changed significantly, as well (P < 0.05 or P < 0.01). The above changes in the liver tissues of F2 generation rats were consistent with the F1 generation rats. Conclusion: Perinatal exposure to 4-NP can affect the synthesis of fatty acid in the livers of F1 and F2 generation rats. The low expression of ERα may be one of the mechanisms by which 4-NP affected fatty acid synthesis in the livers of rats.

Atrazine induced epigenetic transgenerational inheritance of disease, lean phenotype and sperm epimutation pathology biomarkers.

Ancestral environmental exposures to a variety of environmental toxicants and other factors have been shown to promote the epigenetic transgenerational inheritance of adult onset disease. The current study examined the potential transgenerational actions of the herbicide atrazine. Atrazine is one of the most commonly used herbicides in the agricultural industry, in particular with corn and soy crops. Outbred gestating female rats were transiently exposed to a vehicle control or atrazine. The F1 generation offspring were bred to generate the F2 generation and then the F2 generation bred to generate the F3 generation. The F1, F2 and F3 generation control and atrazine lineage rats were aged and various pathologies investigated. The male sperm were collected to investigate DNA methylation differences between the control and atrazine lineage sperm. The F1 generation offspring (directly exposed as a fetus) did not develop disease, but weighed less compared to controls. The F2 generation (grand-offspring) was found to have increased frequency of testis disease and mammary tumors in males and females, early onset puberty in males, and decreased body weight in females compared to controls. The transgenerational F3 generation rats were found to have increased frequency of testis disease, early onset puberty in females, behavioral alterations (motor hyperactivity) and a lean phenotype in males and females. The frequency of multiple diseases was significantly higher in the transgenerational F3 generation atrazine lineage males and females. The transgenerational transmission of disease requires germline (egg or sperm) epigenetic alterations. The sperm differential DNA methylation regions (DMRs), termed epimutations, induced by atrazine were identified in the F1, F2 and F3 generations. Gene associations with the DMRs were identified. For the transgenerational F3 generation sperm, unique sets of DMRs (epimutations) were found to be associated with the lean phenotype or testis disease. These DMRs provide potential biomarkers for transgenerational disease. The etiology of disease appears to be in part due to environmentally induced epigenetic transgenerational inheritance, and epigenetic biomarkers may facilitate the diagnosis of the ancestral exposure and disease susceptibility. Observations indicate that although atrazine does not promote disease in the directly exposed F1 generation, it does have the capacity to promote the epigenetic transgenerational inheritance of disease.

Effect of Gestational Intake of Fisetin (3,3',4',7-Tetrahydroxyflavone) on Developmental Methyl Mercury Neurotoxicity in F1 Generation Rats.

Methyl mercury (MeHg) is a developmental neurotoxin that causes irreversible cognitive damage in offspring of gestationally exposed mothers. Currently, no preventive drugs are established against MeHg developmental neurotoxicity. The neuroprotective effect of gestational administration of a flavanoid against in utero toxicity of MeHg is not explored much. Hence, the present study validated the effect of a bioactive flavanoid, fisetin, on MeHg developmental neurotoxicity outcomes in rat offspring at postnatal weaning age. Pregnant Wistar rats were simultaneously given MeHg (1.5mg/kg b.w.) and two doses of fisetin (10 and 50mg/kg b.w. in two separate groups) orally from gestational day (GD) 5 till parturition. Accordingly, after parturition, on postnatal day (PND) 24, weaning F1 generation rats were studied for motor and cognitive behavioural changes. Biochemical and histopathological changes were also studied in the cerebral cortex, cerebellum and hippocampus on PND 25. Administration of fisetin during pregnancy prevented behavioural impairment due to transplacental MeHg exposure in weaning rats. Fisetin decreased the levels of oxidative stress markers, increased enzymatic and non-enzymatic antioxidant levels and increased the activity of membrane-bound ATPases and cholinergic function in F1 generation rats. In light microscopic studies, fisetin treatment protected the specific offspring brain regions from significant morphological aberrations. Between the two doses of fisetin studied, 10mg/kg b.w. was found to be more satisfactory and effective than 50mg/kg b.w. The present study shows that intake of fisetin during pregnancy in rats ameliorated in utero MeHg exposure-induced neurotoxicity outcomes in postnatal weaning F1 generation rats.

Transgenerational effects of adolescent nicotine exposure in rats: Evidence for cognitive deficits in adult female offspring

This study investigated whether adolescent nicotine exposure in one generation of rats would impair the cognitive capacity of a subsequent generation. Male and female rats in the parental F0 generation were given twice-daily i.p. injections of either 1.0mg/kg nicotine or an equivalent volume of saline for 35days during adolescence on postnatal days 25–59 (P25–59). After reaching adulthood, male and female nicotine-exposed rats were paired for breeding as were male and female saline control rats. Only female offspring were used in this experiment. Half of the offspring of F0 nicotine-exposed breeders and half of the offspring of F0 saline control rats received twice-daily i.p. injections of 1.0mg/kg nicotine during adolescence on P25–59. The remainder of the rats received twice-daily saline injections for the same period. To evaluate transgenerational effects of nicotine exposure on complex cognitive learning abilities, F1 generation rats were trained to perform a highly structured serial pattern in a serial multiple choice (SMC) task. Beginning on P95, rats in the F1 generation were given either 4days of massed training (20patterns/day) followed by spaced training (10 patterns/day) or only spaced training. Transgenerational effects of adolescent nicotine exposure were observed as greater difficulty in learning a “violation element” of the pattern, which indicated that rats were impaired in the ability to encode and remember multiple sequential elements as compound or configural cues. The results indicated that for rats that received massed training, F1 generation rats with adolescent nicotine exposure whose F0 generation parents also experienced adolescent nicotine exposure showed poorer learning of the violation element than rats that experienced adolescent nicotine exposure only in the F1 generation. Thus, adolescent nicotine exposure in one generation of rats produced a cognitive impairment in the next generation.

Effect of Imidacloprid on Reproduction of Female Albino Rats in Three Generation Study

In the present study, the effect of oral administration of imidacloprid over three generations on biochemical, histological and physiological alterations in female rats was assessed. Female rats were divided into three groups. Group 1 was control and was given corn oil, group 2 was administered imidacloprid at the rate of 10 mg/kg bw/day, group 3 was administered imidacloprid at the rate of 20 mg/kg bw/day. F0 and F1 generation female albino rats were dissected for this study. Weight of ovary decreased significantly at higher dose of treated female rats of F0 and F1 generation. Histopathology of ovary of group 2 and group 3 revealed different stages of follicles. The level of acid phosphatase (ACP) and alkaline phosphatase (ALP) increased significantly at higher dose in the ovary of females of both the generations. In either generation, non-significant changes were observed in fertility index, live birth index, gestation lengths and sex ratio. Female F1 pups in the 20 mg/kg/day group showed a significant decreased body weight on postnatal day 21 as compared to F0 pups on day 21. Conclusion: The lower dose of imidacloprid (10 mg/kg/day T1) had no effect on various reproductive parameters of female rats and higher dose (20 mg/kg/day T2) of imidacloprid had some significant effects on feed consumption and reproductive parameters for three generation reproductive study.

Multi-Generations Assessment of Dietary Nucleotides Consumption in Maternal Rats

This study investigated multi-generational reproductive and developmental toxicity of dietary nucleotides (NTs) during lactation in rats with the purpose of augmenting currently available safety data. Weanling rats (F0) were fed NTs at doses of 0.01%, 0.04%, 0.16%, 0.64%, and 1.28% (wt/wt for 90 days and then mated in a 1:1 pattern. The F1 generation rats were fed NTs for 90 days after weaning. The process was repeated until F3 rats were born. We measured the physical and neural development landmarks during lactation in the F3 generation and observed the signs of maternal rats during the gestation, parturition, and lactation periods. No significant differences in either physical or neural developmental landmarks were observed between the control and NT groups. No clinically adverse effects were found in the maternal rats at any point in gestation, parturition, or lactation. There were no alterations in the indicators of parturition. No significant differences were observed between controls and NT-treated rats fed four kinds of nucleotides in F0 milk on PND 21. In conclusion, NTs in doses of up to 1.28% did not cause any adverse effects in either the maternal or filial individuals.

The impact of maternal in utero environment on salt‐induced hypertension in the SS rat (1120.4)

There is considerable interest in mechanisms by which environmental factors impact disease phenotypes later in life. We previously demonstrated that SS (Dahl‐salt sensitive) male rats fed a grain‐based diet (G) show attenuated salt‐induced blood pressure increase and renal damage than animals fed a casein‐based protein diet, AIN‐76a (A). We hypothesized that differences in dietary‐induced susceptibility may be driven by the in utero environment of pregnant females fed these different diets. Using SS rats fed the casein‐based (SSA), or a grain‐based (SSG) diets for many generations, we performed reciprocal embryo transfers, weaned the offspring to a 0.4% NaCl AIN‐76A diet for 6 weeks, then switched the rats to a 4.0% NaCl AIN‐76A diet for mean arterial pressure (MAP) and urinary albumin excretion (UAE) measurements. Compared to control male F1 generation rats generated by the transfer of SSA embryos into SSA females (SSA&gt;A), F1 males generated by transfer of SSA embryos into SSG female recipients (SSA&gt;G) demonstrated significantly reduced pressure (MAP, 157±6 mmHg vs. 138±4; n=8‐14/group) and renal damage (UAE, 204±16 mg/day vs. 96±13 mg/day; n=12‐18/group) after 21 days of exposure to a 4.0% NaCl diet. Conversely, reciprocal embryo transfer of SSG embryos into SSA recipients (SSG&gt;A) promoted hypertension (MAP, 166±5 mmHg, n=21) and renal disease (UAE, 187±27 mg/day, n=25) in male offspring. These data support the hypothesis that the maternal in utero environment is a significant driving factor in the hypertensive phenotype of the SS rat. Interestingly, following breeding the of the protected SSA&gt;G F1 animals on the A diet, the disease attenuation was lost in the F2 generation (MAP=152±5 mmHg; Ualb=145±25 mg/day, n=12‐13/group), suggesting that; 1) that the protective effect of the G diet is not transgenerational and 2) that severe salt‐induced hypertension disease phentoypes can be restored to SS rats fed the grain‐based diet by switching the breeders to the pro‐hypertensive AIN‐76A diet.

Plastics Derived Endocrine Disruptors (BPA, DEHP and DBP) Induce Epigenetic Transgenerational Inheritance of Obesity, Reproductive Disease and Sperm Epimutations

Environmental compounds are known to promote epigenetic transgenerational inheritance of adult onset disease in subsequent generations (F1–F3) following ancestral exposure during fetal gonadal sex determination. The current study was designed to determine if a mixture of plastic derived endocrine disruptor compounds bisphenol-A (BPA), bis(2-ethylhexyl)phthalate (DEHP) and dibutyl phthalate (DBP) at two different doses promoted epigenetic transgenerational inheritance of adult onset disease and associated DNA methylation epimutations in sperm. Gestating F0 generation females were exposed to either the “plastics” or “lower dose plastics” mixture during embryonic days 8 to 14 of gonadal sex determination and the incidence of adult onset disease was evaluated in F1 and F3 generation rats. There were significant increases in the incidence of total disease/abnormalities in F1 and F3 generation male and female animals from plastics lineages. Pubertal abnormalities, testis disease, obesity, and ovarian disease (primary ovarian insufficiency and polycystic ovaries) were increased in the F3 generation animals. Kidney and prostate disease were only observed in the direct fetally exposed F1 generation plastic lineage animals. Analysis of the plastics lineage F3 generation sperm epigenome previously identified 197 differential DNA methylation regions (DMR) in gene promoters, termed epimutations. A number of these transgenerational DMR form a unique direct connection gene network and have previously been shown to correlate with the pathologies identified. Observations demonstrate that a mixture of plastic derived compounds, BPA and phthalates, can promote epigenetic transgenerational inheritance of adult onset disease. The sperm DMR provide potential epigenetic biomarkers for transgenerational disease and/or ancestral environmental exposures.

Dioxin (TCDD) Induces Epigenetic Transgenerational Inheritance of Adult Onset Disease and Sperm Epimutations

Environmental compounds can promote epigenetic transgenerational inheritance of adult-onset disease in subsequent generations following ancestral exposure during fetal gonadal sex determination. The current study examined the ability of dioxin (2,3,7,8-tetrachlorodibenzo[p]dioxin, TCDD) to promote epigenetic transgenerational inheritance of disease and DNA methylation epimutations in sperm. Gestating F0 generation females were exposed to dioxin during fetal day 8 to 14 and adult-onset disease was evaluated in F1 and F3 generation rats. The incidences of total disease and multiple disease increased in F1 and F3 generations. Prostate disease, ovarian primordial follicle loss and polycystic ovary disease were increased in F1 generation dioxin lineage. Kidney disease in males, pubertal abnormalities in females, ovarian primordial follicle loss and polycystic ovary disease were increased in F3 generation dioxin lineage animals. Analysis of the F3 generation sperm epigenome identified 50 differentially DNA methylated regions (DMR) in gene promoters. These DMR provide potential epigenetic biomarkers for transgenerational disease and ancestral environmental exposures. Observations demonstrate dioxin exposure of a gestating female promotes epigenetic transgenerational inheritance of adult onset disease and sperm epimutations.

Biochemical and reproductive effects of gestational/lactational exposure to lead and cadmium with respect to testicular steroidogenesis, antioxidant system, endogenous sex steroid and cauda-epididymal functions

This study investigated the effects of gestational and lactational exposure to lead and cadmium on testicular steroidogenesis, antioxidant system and male accessory gland functions in F1 generation rats to understand the biochemical mechanisms involved in endocrine disruptions. Pregnant rats were subcutaneously administered with 0.05 mg kg(-1) body wt\ day(-1) of sodium acetate (control), lead acetate, cadmium acetate and (lead acetate + cadmium acetate) throughout the gestational-lactational period, and all animals from each of the experimental groups were sacrificed by decapitation on post-natal day 56 for performing various biochemical assays. We observed significant reduction in the activities of testicular key steroidogenic enzymes and serum testosterone concentration along with significant depletion in cholesterol, ascorbic acid and reduced glutathione contents in all the metal-treated groups. Reductions in the activities of catalase and superoxide dismutase with concomitant increase in the levels of thiobarbituric acid reactive substance were observed in experimental groups. Both sperm contents and sperm motility patterns were significantly altered in all the metal-treated groups, suggesting the direct/indirect spermotoxic effects of lead and cadmium. The inhibitory effects of lead, cadmium and combined exposure on testicular steroidogenesis machinery, along with the male accessory gland functions, are indicative of multiple targets of lead and cadmium to disrupt male reproductive functions.

Biochemical and molecular effects of gestational and lactational coexposure to lead and cadmium on ovarian steroidogenesis are associated with oxidative stress in f1 generation rats

Few studies have characterized the molecular and biochemical mechanisms involved in ovarian steroidogenesis disruption by heavy metals, such as lead and cadmium coexposure, on F1 generation offspring. In this study, adult pregnant female rats were treated subcutaneously (0.05 mg/kg of body weight per day) with sodium acetate (control), lead acetate, and cadmium acetate separately and in combination throughout gestational and lactational period, and all animals from each of the experimental groups were sacrificed by decapitation on postnatal day 56 for various assays. The activities of key steroidogenic enzymes (17β-hydroxysteroid dehydrogenase and 3β-hydroxysteroid dehydrogenase) decreased in all the metal-treated groups. But the most significant decrease in the activities was observed in the cadmium-treated group, whereas the combined exposure group showed an intermediate effect. Serum estradiol and progesterone levels were also significantly altered in all the metal-treated groups, with the cadmium-exposed group showing maximum reductions as compared with the control group. The inhibitory effects of lead and cadmium on ovarian steroidogenic acute regulatory protein (StAR) mRNA levels along with CYP11 mRNA levels were also observed. Ovarian cholesterol content measured also showed significant depletion in all the metal-treated groups, with the cadmium-exposed group showing the maximum depletion. The activities of ovarian enzymatic antioxidants, such as superoxide dismutase, catalase, and glutathione peroxidase, were all significantly diminished along with significant depletion in nonenzymatic antioxidants. Lipid peroxidation was elevated significantly in all the metal-treated groups. In conclusion, lead and cadmium inhibit ovarian steroidogenesis by downregulating StAR gene expression along with inhibiting activities of steroidogenic enzymes and antioxidant system.

The effects of bisphenol A on sex hormone levels of F0 female rats and F1 male rats during weaning period

The effects of bisphenol A (BPA) were examined on sex hormones of F1 generation male rats during weaning period. Female rats were exposed to BPA from day 0 after pregnancy to the weaning period at doses of 50, 100, or 200 mg kg−1. The sex hormone levels of F1 generation male rats were determined. This study shows that F0 generation female rats fed with 200 mg kg−1 BPA had a significantly higher serum prolactin (PRL) levels at the end of weaning. Significantly higher levels of serum estradiol (E2) were also found in female rats fed 100 or 200 mg kg−1 BPA. Serum levels of E2 in F1 male generation rats were higher in treatment groups compared to control groups while serum testosterone (T) levels were lower. Follicle stimulating hormone (FSH) in F1 generation rats fed 200 mg kg−1 was markedly decreased. The relative testicular weights were significantly less in 100 and 200 mg kg−1 BPA groups. BPA was found to alter the sex hormone levels in F1 male rats during weaning period and thus disrupted endocrine functions.

Epigenetic Transgenerational Actions of Vinclozolin on Promoter Regions of the Sperm Epigenome

Previous observations have demonstrated that embryonic exposure to the endocrine disruptor vinclozolin during gonadal sex determination promotes transgenerational adult onset disease such as male infertility, kidney disease, prostate disease, immune abnormalities and tumor development. The current study investigates genome-wide promoter DNA methylation alterations in the sperm of F3 generation rats whose F0 generation mother was exposed to vinclozolin. A methylated DNA immunoprecipitation with methyl-cytosine antibody followed by a promoter tilling microarray (MeDIP-Chip) procedure was used to identify 52 different regions with statistically significant altered methylation in the sperm promoter epigenome. Mass spectrometry bisulfite analysis was used to map the CpG DNA methylation and 16 differential DNA methylation regions were confirmed, while the remainder could not be analyzed due to bisulfite technical limitations. Analysis of these validated regions identified a consensus DNA sequence (motif) that associated with 75% of the promoters. Interestingly, only 16.8% of a random set of 125 promoters contained this motif. One candidate promoter (Fam111a) was found to be due to a copy number variation (CNV) and not a methylation change, suggesting initial alterations in the germline epigenome may promote genetic abnormalities such as induced CNV in later generations. This study identifies differential DNA methylation sites in promoter regions three generations after the initial exposure and identifies common genome features present in these regions. In addition to primary epimutations, a potential indirect genetic abnormality was identified, and both are postulated to be involved in the epigenetic transgenerational inheritance observed. This study confirms that an environmental agent has the ability to induce epigenetic transgenerational changes in the sperm epigenome.

Peri- and postnatal rodent development of Hematide, an erythropoiesis-stimulating agent

Aperi- and postnatal reproduction toxicity study was conducted in rats treated with Hematide, a synthetic PEGylated peptidic erythropoiesis stimulating agent (ESA). Hematide, at IV doses of 0, 0.5, 3, and 15 mg/kg, was administered from implantation through lactation on gestation days (GDs) 5 and 18 and lactation day (LD) 13. Hematide induced pronounced polycythemia in all Hematide-treated dams. On LDs 2 and 21, hemoglobin (Hgb) increases above control levels were 3.1, 5.2, and 5.0 g/dL and 4.1, 5.1, and 5.5 g/dL at the 0.5, 3, and 15 mg/kg/dose, respectively. There were no effects on parturition, lactation, or maternal behavior in the F0 generation female rats. A slight decrease in pup viability on postpartum days 2-4 and lower body weights and/or body weight gain for the F1 generation were associated with pronounced polycythemia and decreases in maternal body weight gain and/or food consumption at > or =3 mg/kg/dose. Hematide fetal exposure was negligible. No Hematide effect, other than on growth and survival, was noted on developmental, functional, mating, and fertility end points in the F1 generation rats, and no effect on litter or fetal parameters was observed in the F2 generation. The maternal no-observed-adverse-effect level (NOAEL) for Hematide was 0.5 mg/kg, and the NOAEL for parturition and maternal behavior was 15 mg/kg. The NOAEL for F1 pup viability and growth was 0.5 mg/kg/dose. In conclusion, the Hematide-associated adverse findings were attributed to exaggerated erythropoiesis (pronounced and prolonged polycythemia) resulting from administration of an ESA to pregnant animals.

Effects of nonylphenol on brain gene expression profiles in F1 generation rats

The effects of nonylphenol (NP) were examined on brain gene expression profiles of F1 generation rats by means of microarray techniques. The mRNA were extracted from the brain of two-day-old F1 generation rats whose F0 male generation were treated with NP, then reversely transcribed to cDNA and labeled with Cy5 and Cy3 fluorescence. Subsequently, the cDNA probes were hybridized to the mouse 40S cDNA microarray and the fluorescent signals of Cy5 and Cy3 were scanned and analyzed. Sixteen identified genes were expressed significantly differently from control, including 13 down-regulated, of which five were related to brain energy metabolism. Data suggest that it is possible that NP affects function of energy metabolism in male rats when administered perinatally.

Transgenerational hormonal imprinting caused by vitamin A and vitamin D treatment of newborn rats. Alterations in the biogenic amine contents of the adult brain

Biogenic amines (norepinephrine, dopamine, homovanillic acid, serotonin and 5-hyroxyindole acetic acid) were measured by HPLC method in adult F1 generation rats’ brain regions (brainstem, hypothalamus, hippocampus, striatum and frontal cortex), whose mothers (P generation) were treated with vitamin A or vitamin D neonatally (hormonal imprinting). Many significant differences were found, related to the maternally untreated controls. In the earlier studied P generation females, vitamin A consistently influenced the serotonerg system (5HIAA), while vitamin D the dopaminerg system (DA or HVA). Vitamin A imprinting always resulted in reduced, while that by vitamin D always in increased tissue levels. In the present case (directly untreated F1 generation) the transgenerational effect was not unidirectional, however biogenic amine tissue levels were strongly disturbed and brain-area dependent. The results call attention to the transgenerational effect of hormonal imprinting in the case of receptor level acting vitamins which are frequently used in the most imprinting-sensitive period (perinatally) of human life and suggests that caution is warranted.

Effects of nonylphenol on endocrine regulation-associated gene expression profiles in whole brain of F1 generation male rats

In order to determine the effects of nonylphenol (NP) on endocrine regulation-associated gene expression profiles in whole brain of F1 generation rats, mRNA extracted from the brain of 2-day old F1 generation rats whose F0 male generation was treated with NP. The mRNA was then reversely transcribed to cDNA and labeled with cy5 and cy3 fluorescence. Subsequently, cDNA probes were hybridized to the Mouse40S cDNA microarray and the fluorescent signals of cy5 and cy3 were scanned and analyzed. Sixteen genes were identified which expressed differently, including 13 that were down-regulated in which 4 were related to brain regulation of endocrine function. Data suggest that NP mainly affects metabolism and synthesis of steroid hormone in various ways and disturbs the reproductive function of male rats when administered perinatally.

Developmental and reproductive outcome of F1 generation of Wistar rats exposed to Acanthus montanus aqueous extract during organogenesis

Acanthus montanus (Acanthaceae) leaves aqueous extract (AMAE) is used against threatened abortion and pain, thus, its potential adverse effects on pregnancy and postnatal development of F1 generation rats needed investigation. Pregnant Wistar rats (170–210g) received by gavage, graded doses of AMAE (0, 62.5, 125, 500 and 1000mg/kg/day) from days 6–15 of gestation. Earlier, we have studied the effect of its Methanol/Methylene Chloride extract on pregnant rats [1]. Daily maternal behavioural changes, body weights, food and water intakes were noted. On delivery, the progeny were evaluated at birth through lactation to weaning for changes in litter size, mortality, fetal body weight, crump-rump and tail lengths, developmental landmarks, behavioural functions, food and water consumptions, pups reproductive capacities, external, visceral and skeletal malformations. One-way ANOVA and Dunnett's multicomparison post-test at P&lt;0.05 were used. Maternal weight loss was absent and unaccompanied with toxicity, food and water intake changes. Litter size and sex ratio were unaffected. Fetal body weights, crown-rump and tail lengths, food and not water consumptions were dose independently decreased. Reduced survival and lactation indices, delayed eye openings, limb deformations, inflamed eyes and hair loss occurred with impairment of some behavioural function tests. Necropsy findings revealed reduced skin and organ fats. No skeletal anomaly was encountered. All exposed offsprings were infertile. AMAE is a developmental toxicant, thus, its use in the first three months of pregnancy should be avoided.

Effects of Nonylphenol on Brain Gene Expression Profiles in F1 Generation Rats

To explore the effects of nonylphenol on brain gene expression profiles in F1 generation rats by microarray technique. mRNA was extracted from the brain of 2-day old F1 generation male rats whose F0 female generation was either exposed to nonylphenol or free from nonylphenol exposure, and then it was reversely transcribed to cDNA labeled with cy5 and cy3 fluorescence. Subsequently, cDNA probes were hybridized to two BiostarR-40S cDNA gene chips and fluorescent signals of cy5 and cy3 were scanned and analyzed. Results Two genes were differentially down-regulated. Nonylphenol may disturb the neuroendocrine function of male rats when administered perinatally.