Development In Hamsters Research Articles

A Physiologically Based Pharmacokinetic Model for Arsenic Exposure: I. Development in Hamsters and Rabbits

A physiologically based pharmacokinetic model for exposure to inorganic arsenic in hamsters and rabbits has been developed. The model in its present state simulates three routes of exposure to inorganic arsenic: oral intake, intravenous injection, and intratracheal instillation. It describes the tissue concentrations and the urinary and fecal excretions of the four arsenic metabolites: inorganic As(III) and As(V), methylarsonic acid, and dimethylarsinic acid. The model consists of five tissue compartments, chosen according to arsenic affinities: liver, kidneys, lungs, skin, and others. The model is based on physiological parameters, which were scaled according to body weight. When physiological parameters were not available, the data for the model were obtained by fitting (tissue affinity, absorption rate, and metabolic rate constants). The excretions of the arsenic metabolites in urine and feces are well simulated with the model for both species. Further validation of the arsenic metabolite concentrations in the tissues andin vitromeasurements of the tissue affinity constants are discussed.

Influence of Prenatal and Postnatal Photoperiods on Postnatal Testis Development in the Siberian Hamster (Phodopus Sungorus)1

In Siberian hamsters, juvenile testicular development is regulated to a large extent by photoperiod. Mother hamsters are able to pass photoperiodic information to their male fetuses, and this information can influence their postnatal gonadal development. In this study, we investigated the effects of gestation in long (16L:8D) and short (10L:14D) day lengths on the rates of juvenile testicular growth in several different postnatal photoperiods. On the day of parturition, parents and young from each gestation photoperiod were raised in 14L:10D through Day 13 of life and then were exposed to one of six photoperiods--16L:8D, 15L:9D, 14L:10D, 13L:11D, 12L:12D, or 10L:14D--until Day 32 of age. The data indicated that 15L and 14L are the minimal day lengths required to prevent complete inhibition of testis growth in long (16L) and short day (10L)-gestated hamsters, respectively. These results support earlier findings suggesting that gestation photoperiod can influence the rate of reproductive development in a certain range of "intermediate" postnatal day lengths (14L to 15L), but that gestation photoperiod does not alter the pattern of testis development in hamsters exposed to other (i.e., longer or shorter) postnatal photoperiods. Thus, both the absolute length and the direction of change of photoperiods experienced in early life can influence prepubertal testis growth.

Effects of photoperiod, pinealectomy, and melatonin implants on testicular development in juvenile Siberian hamsters (Phodopus sungorus).

When transferred from a long to short photoperiod, Siberian hamsters (Phodopus sungorus) undergo a number of physiological and morphological changes including suppression of gonadal activity, a change in pelage color, a decrease in body weight, and, in response to a simultaneous decrease in ambient temperature, physiological changes associated with the induction of daily torpor. All these functions can be affected by photoperiod and melatonin treatment. To investigate the interactive effects of photoperiod, pinealectomy, and melatonin on gonadal development, two experiments were performed using juvenile Siberian hamsters. In experiment 1, animals born in a long photoperiod (16L:8D) either remained in a long photoperiod or were transferred to a short photoperiod (8L:16D) from 15 days of age, when surgeries (pinealectomy and/or melatonin implantation) were performed. Testicular development was inhibited in all animals bearing melatonin implants irrespective of the presence or absence of the pineal gland. Pinealectomy blocked the inhibitory effect of short photoperiod on maturation of the reproductive system. Therefore, the pineal gland must be involved in the short photoperiod- induced inhibition of testicular maturation of juvenile Siberian hamsters. In experiment 2, a similar experimental design was employed except that the hamsters were born and raised to 15 days of age in 8L:16D. Exogenous melatonin, pinealectomy, or both retarded gonadal development in hamsters born in 8L:16D and transferred on Day 15 of age to 16L:8D. All hamsters maintained in a short photoperiod had small testes irrespective of the presence of absence of the pineal gland or of melatonin implants. Hamsters transferred to a long photoperiod after pinealectomy and/or melatonin implantation had small testes compared to those of the control group.(ABSTRACT TRUNCATED AT 250 WORDS)

Development of fungiform papillae, taste buds, and their innervation in the hamster.

Fungiform taste buds in mature hamsters are less subject to neurotrophic influences than those of other species. This study evaluates taste-bud neurotrophism during development in hamsters by examining the relation between growing nerves and differentiating fungiform papillae. Chorda tympani (CT) or lingual (trigeminal) nerve (LN) fibers were labelled with Lucifer Yellow as they grew into (CT fibers) or around (LN fibers) developing taste buds. Developing fungiform papillae and taste pores were counted with the aid of a topical tongue stain. The tongue forms on embryonic days (E) 10.5-11 and contains deeply placed CT and LN fibers but no papillae. By E12, the tongue epithelium develops scattered elevations. These "eminences" selectively become innervated by LN fibers that grow to the epithelium earlier and in larger numbers than CT fibers. Definitive fungiform papillae form rapidly during E13-14 and become heavily innervated by LN fibers. Intraepithelial CT fibers, rare at E13, invariably innervate fungiform papillae containing nascent taste buds at E14. During E14-15 (birth = E15-16), most papillae contain taste buds with pores, extensive perigemmal LN innervation, and extensive intragemmal CT innervation. At birth, numbers of fungiform papillae and taste pores are adultlike. The results show that fungiform eminences begin forming in the absence of innervation. The subsequent differentiation of definitive fungiform papillae and their innervation by LN fibers occur synchronously, prior to the differentiation of taste buds and their CT innervation. The hamster is precocious (e.g., compared to rat) in terms of LN development and the structural maturity of the anterior tongue at birth.

Inhibition of in vitro fertilization and early embryonic development in hamsters by gossypol

We have previously reported an inhibitory effect of gossypol and its metabolite on bovine and mouse early embryonic development. In the present study, eggs were collected from oviducts of superovulated hamsters. Epididymal sperm were used for in vitro fertilization (IVF). Gossypol at 5, 10, and 30 μg/ml significantly inhibited the formation of 2 pronuclei by 45, 65 and 95%, respectively. On the first day of pregnancy, hamsters were given an intrauterine treatment of 200 μg of gossypol in 100 μl of corn oil per uterine horn. On day 3, embryos from controls were in morula (65%) and early morula (17%) stages, while less than 2% of embryos from the gossypol-treated hamsters were in the morula stage The numbers of embryo implantation sites on day 8 and pups in controls (14 ± 2.0 and 12 ± 1.5, respectively) were significantly higher than those in the gossypol-treated hamsters (8.5 ± 20 and 4.0 ± 1.5, respectively). Our results suggest that gossypol is able to affect fertilization, embryonic development, embryo implantation, and the number of pups in hamsters through a not-yet-defined mechanism.

Energy substrate requirements for in-vitro development of hamster 1- and 2-cell embryos to the blastocyst stage

Energy substrate requirements (pyruvate, lactate and amino acids) were determined for in-vitro development of hamster 1- and 2-cell embryos to blastocysts, using a chemically defined, protein free medium (hamster embryo culture medium, HECM). One-cell embryos were very sensitive to energy substrate type and concentration. Pyruvate alone could not support development of 1-cell embryos to greater than 4-cells, whereas lactate as sole energy substrate supported 14% development into morulae/blastocysts. Pyruvate, with lactate and 20 amino acids, inhibited 1-cell embryo development into blastocysts relative to lactate and 20 amino acids. The highest development of 1-cell embryos to blastocysts (up to 27%) occurred with reduced lactate concentration (less than 10 mM) and either 20 amino acids or 0.2 mM glutamine. Hamster 2-cell embryos were much less sensitive to energy substrates, requiring only lactate for development to blastocysts (53%). Lactate with 20 amino acids supported 70-75% of 2-cell embryos to blastocysts. Glutamine as sole energy and nitrogen source supported development to morulae and blastocysts of some 2-cell, but not 1-cell, embryos. Pyruvate did not enhance development of 2-cell embryos. We conclude that (i) altering the types and concentrations of available energy substrates drastically changes the developmental responses of 1-cell hamster embryos in vitro and (ii) energy substrate requirements for hamster embryo development in vitro are markedly different from those of mouse embryos, the standard model for studies on preimplantation development. This is the first report of successful in-vitro culture of hamster 1-cell embryos to the blastocyst stage.

Influence of single amino acids on the development of hamster one‐cell embryos in vitro

One-cell hamster embryos placed in culture have always shown a complete block to development at the two-cell stage. In a preliminary study using a chemically defined culture medium containing 20 amino acids (HECM-1), many one-cell embryos were able to escape the "two-cell block" and develop to the four-cell stage. Use of a simpler formulation containing only the amino acids hypotaurine and glutamine revealed marked inhibitory and stimulatory effects of adding the other amino acids. In the first experiment, 19 amino acids were separately examined for effects on one-cell embryo development. Six amino acids (phenylalanine, valine, isoleucine, tyrosine, tryptophan, and arginine) inhibited embryo development (reduced mean cell number; MCN), and three others (glycine, cystine, and lysine) stimulated development (increased MCN), compared with basic medium containing only glutamine and hypotaurine (low control). When the responses with the six inhibitory amino acids were totalled, only 3 of 185 (2%) one-cell embryos reached the six-or seven-cell stage compared to a total of 15 of 76 (20%) embryos that developed to these stages using the three stimulatory amino acids. When tested together in a second experiment, the six inhibitory amino acids significantly reduced the MCN, from 4.28 +/- 0.44 (low control) to 3.71 +/- 0.55. In this group, 17 of 117 (15%) of one-cell embryos reached more than four-cell and only 4 of 117 (3%) reached six- or 7-cell stages, compared with 39 of 117 (33%) and 12 of 117 (10%), respectively, for the basal medium group.(ABSTRACT TRUNCATED AT 250 WORDS)

Elevated activity of pancreatic type amylase in the urine as an early indicator of pancreatic tumors in hamsters

Levels of amylase isozymes in the urine and serum of Syrian hamsters during pancreatic carcinogenesis, induced by N-nitrosobis(2-oxopropyl)amine (BOP), were investigated. BOP was injected subcutaneously (sc) into female Syrian golden hamsters at a dose of 10mg/kg once weekly for 6 wk and amylase activities in urine and serum samples were measured every other week from the first treatment of BOP. Although total amylase in the urine showed no remarkable changes, the pancreatic type isozyme demonstrated only very low levels for the first 6 wk and then from wk 8, became elevated showing continuously high levels in all animals thereafter. Animals were sacrificed at wk 10, 14, and 18. Dysplastic lesions of the pancreas developed in all the hamsters investigated. Furthermore, pancreatic adenocarcinomas were also observed in all animals sacrificed at wk 18. Thus, the results suggest that measurement of pancreatic isoamylase in the urine might allow early indication of pancreatic tumor development in hamsters.

Direct effects of retinoic acid on the development of the tail bud in chick embryos.

Retinoic acid (RA) has been reported to induce vascular lesions and haematoma formation in the vicinity of the tail bud during the critical period for inducing abnormalities of tail bud development in hamsters (Wiley, '83; Tibbles and Wiley, '88), mice (Tibbles and Wiley, '88) and chicken embryos (Jelinek and Kistler, '81). Experiments were conducted to determine whether or not these vascular lesions were the primary cause of the malformations which they accompanied. Chick embryos were exposed for varying lengths of time to several dosages of RA. Primitive streaks or tail buds from treated embryos were then excised prior to vascularization and transplanted to the coelomic walls of untreated host embryos. The grafts were harvested at 3 or 6 days after grafting and processed for histological examination. Observations of serial sections of controls showed that the primitive streak and early (stage 13-14) tail bud were able to form neural tubes and a variety of other structures including ganglia, nerve fibres, and kidney tubules. Treatment of donor embryos with RA prior to grafting, however, affected the frequency and characteristics of the neural tubes and other tissues developing in the grafts. The effects of RA on development were correlated with both the dosage and length of exposure to the teratogen prior to grafting. Since the grafts were made before the appearance of blood vessels in the tail buds, we have concluded that the effects of RA on the development of tail bud tissues, and especially the secondary neural tube, are direct and are not mediated solely through the disruptive effects of vascular lesions seen in intact embryos.

Effect of DMBA on oral cancer development in hamsters with latent HSV-1 infections in trigeminal ganglia

This study examined the effect of 7, 12-dimethyl-benz(a)anthracene (DMBA), polycyclic aromatic hydrocarbon carcinogen, on pathologic changes in buccal pouch mucosa of hamsters with latent herpes simplex virus type 1 (HSV-1) infections in their trigeminal ganglia. Of the pouches receiving DMBA treatment, the average number of tumors per pouch was significantly higher ( p < 0.01) in pouches of animals with HSV-1 infections as compared to those that received DMBA only. Of the pouches receiving DMBA, the average cumulative tumor diameter (the sum of the tumor diameters in each group divided by the number of pouches in that group) was notably greater in animals with latent HSV-1 than in animals that had not been infected; however, this difference was not significant. The DMBA-treated hamsters that had latent infections also displayed a higher severity and prevalence of histopathologic changes in their pouch mucosa. This study indicates that latent HSV-1 and DMBA show synergism in the development of tumors and precancerous histopathologic changes in hamster buccal pouch mucosa. An unexpected finding was that among the HSV-1 infected animals, 95% of the ganglia from animals treated with DMBA showed latent HSV-1 virus on explanation culture, whereas only 10% of the ganglia from infected animals that received mineral oil, rather than DMBA, contained latent virus.

The effect of cadmium on the development of the facial prominences: Surface area measurements of day 10‐8 A.M. hamster embryos

The environmental contaminant cadmium (Cd) is a proven teratogenic agent in rodents. In hamsters, it causes craniofacial dysmorphogenesis. The underlying mechanism for this damage is unknown. Early facial development in hamsters occurs during gestation days 9-11 and involves the formation and appropriate fusion of several prominences surrounding the stomodeum. The hypothesis for this study is that the occurrence of Cd-induced facial defects involves a disruption of the normal formation and/or fusion of one or more of the facial prominences. Pregnant hamsters were treated with Cd (2 mg/kg) or water intravenously on gestation day 8 (8 A.M.). On gestation day 10 (8 A.M.) surviving embryos were processed to obtain scanning electron micrographs of the frontal view of the face. Measurements of the surface areas of 15 different portions of the face were obtained using a microcomputer equipped with a digitizer. Both qualitative and quantitative differences in the faces were detected upon comparing the Cd-exposed and control embryos. The surface areas of the prominences measured were significantly smaller in the Cd-exposed embryos. However the sizes of the other regions of the Cd-exposed faces were either little affected (nasal pit areas) or markedly increased (the interval of the face between the medial nasal prominences). Two possible explanations for these data are discussed.

Normal development of hamster and rabbit eggs fertilized by spermatozoa labelled with the fluorescent thiol alkylating agent, monobromobimane.

Cauda epididymal spermatozoa of the golden hamster were labelled with the thiol-alkylating reagent, monobromobimane (MB). Female hamsters underwent uterine insemination with labelled spermatozoa at laparotomy under metofane anesthesia. All 12 females examined between 5 and 54 h postinsemination yielded a total of 83/100 (83%) eggs in the process of fertilization or embryos. Under ultraviolet (UV) exposure all exhibited a fluorescent tail which, in the 4- and 8-cell embryos, could be seen to be fraying or disintegrating. As cleavage progressed, labelled tail components came to be restricted among the blastomeres such that at the 4- and 8-cell stage the tail could be seen in only one to three blastomeres. To study complete development and pregnancy another 12 females received uterine insemination. After recovery from anesthesia (approximately equal to 4 h) these females were mated with a vasectomized male bearing a dominant genetic marker (black eyes) to allow unequivocal determination of paternity in the fetuses and young produced. Seven became pregnant with one female losing her pregnancy about Day 7 of gestation. Two females sacrificed on Day 13 produced 17 normal fetuses and one resorption. Four females delivered 16 young, all normal at birth and in subsequent growth and fertility. In addition, insemination of female rabbits with MB-labelled spermatozoa yielded normal embryos (50/52 96%) from 3 does on Day 2 and (38/64 60%) from 4 does on Days 4 or 5. Two normal litters (9 bunnies) have delivered from 3 does allowed to carry to term.(ABSTRACT TRUNCATED AT 250 WORDS)

Ultrastructural and cytochemical observations on 5-fluorouracil-induced cleft-palate development in hamster.

Sequential alterations in 5-fluorouracil-treated hamster fetal palate were studied by light and electron microscopy and by acid phosphatase cytochemistry. At an early stage in 5-fluorouracil-treated fetuses, when the palatal shelves were vertical, lysosomes first appeared in cells of the prospective fusion epithelium and then in the cells of subjacent mesenchyme. In contrast to controls, increasing numbers of both the epithelial and mesenchymal cells of the vertical palate showed lysosomal injury in 5-fluorouracil-treated fetuses as development progressed. Subsequently, the basal lamina in the vertical palate showed alterations, characterized initially by disturbances in lamina lucida, by fingerlike extensions of lamina densa, and ultimately by its complete breakdown. At a later stage, when shelves became horizontal, the lysosomes were absent in both the epithelial and mesenchymal cells, and the basal lamina continuity was restored. Unlike controls, however, 5-fluorouracil-treated horizontal shelves never contacted one another. Instead, the epithelia of the horizontal shelves underwent stratification. It appears that premature formation of lysosomes in palatal epithelial and mesenchymal cells following 5-fluorouracil treatment disrupts normal cytodifferentiation and affects the integrity of the basal lamina; both effects are associated with cleft-palate development.

Pregnancy and fetal development in hamsters treated with prostaglandin F 2α

The influence of PGF 2α on pregnancy and fetal outcome was investigated in the hamster. Following subcutaneous treatment with 50, 100, 200, 400, and 800 μg PGF 2α prenatal loss was significantly increased only at the highest dose level. The offspring of the treated animals were all alive and normal. Fetal weight was not affected. However, following intravenous injection of 100 and 200 μg PGF 2α there was a significant reduction in fetal weight, and at the 400 μg dose level an increase in fetal resorptions. All fetuses recovered were alive and normal. These results indicate that PGF 2α is not teratogenic in hamsters despite the apparent greater sensitivity of the hamster embryo to prostaglandin.

The distribution of melanin in the developing optic cup and stalk and its relation to cellular degeneration

The early distribution of melanin in the developing optic cup and stalk and its relationship to cellular degeneration have been studied during intrauterine development in hamsters, mice, ferrets, and humans. The extensive degenerative changes that occur in the region of the optic fissure do not involve any pigment-bearing cells. Melanin is not formed at the site of the fissure until after the processes of fusion and the associated degenerative changes are complete. In contrast to this, there is a small region near the junction of the cup and stalk where melanin is particularly related to degenerative changes. Melanosomes form dense clumps associated with basophilic, apparently degenerate areas of cytoplasm, and these melanosome complexes themselves undergo further degenerative changes. The two types of degenerative changes, pigmented and unpigmented, are limited to the same two regions of the developing eye in all four species. However, the pigment degeneration is more extensive in ferrets and humans than in mice and hamsters. These observations raise the possibility that the lysis of melanosomes plays a significant but undefined local role in ocular development.

Failure of two retinoids to inhibit tracheal carcinogenesis in hamsters

The effect of 2 retinoids, 13-cis-retinoic acid and 4-methoxy-2,3,6-trimethylphenyl analog of retinoic acid ethyl amide (designated Roll-1430), on tracheal tumor development in hamsters exposed to N-nitroso-N-methylurea was tested. Hamsters were intratracheally exposed either 18, 20, or 23 times to 1% N-nitroso-N-methylurea before the retinoids were administered in the diet. Evidence was presented which indicates that the great majority of the animals are free of invasive neoplasia at the start of retinoid feeding. In none of the 6 retinoid-treated groups could a statistically significant inhibition of tumor development be demonstrated. Hamsters treated with 13-cis-retinoic acid (128 or 172 mg/kg of diet) tended to have an elevated cancer risk; this effect was at a statistically significant level in the group treated with 172 mg/kg of diet. The distribution of histologic tumor types seemed to be shifted in favor of adeno and mixed adeno-epidermoid tumors in the group receiving a low carcinogen dose and Roll-1430. Similar to earlier studies with retinyl acetate tested in hamsters and rats, 13-cis retinoic acid and the retinoic acid ethyl amide analog Roll-1430, failed to inhibit development of respiratory tract neoplasms. We suspect that the reason for this is the absence of significant promoting influences in the current lung cancer models and that retinoids act mostly as anti-promoters.

Dipetalonema viteae: Primary, secondary and tertiary infections in hamsters

Hamsters were given primary infections of 100, 200, and 300 D. viteae larvae and groups killed at various intervals after infection. In addition, hamsters were sequentially infected with 100, 200, and 300 larvae and groups killed at 100 or 75 days after the secondary and tertiary infection, respectively. Blood microfilariae were detected on Day 60 following a primary infection, reached a maximum on Day 75, declined to low levels by Day 105, and were negative on Day 120. No microfilariae reappeared in the blood of hamsters given secondary or tertiary infections. Between 20–30% of the infecting larval dose had reached the adult stage by Days 75 or 100 postinfection in hamsters given primary, secondary, or tertiary infections. There was no evidence of arrested larval development in hamsters receiving a second or third challenge infection. Almost half of the tertiary infection hamsters developed subcutaneous nodules and their numbers varied greatly among individual animals. The nodules variously contained living worms, pus, and fragmented worms, or pus only. Hamsters given primary infections of 100, 200, or 300 larvae and killed 375 days after infection had no subcutaneous nodules; however, hamsters given the 200 and 300 larval infections were seen to have dead worms in the subcutaneous tissues. No stunting of adult worms was noted and all female worms had uteri packed with microfilariae.

Development in Hamsters of Antibodies against Surface Antigens Present in Cells Transformed by Papovavirus SV40

Summary Newborn hamsters inoculated with SV40 and later challenged with the homologous virus in the latent period developed antibodies which reacted specifically with antigens at the surface of hamster cells transformed by SV40. Regression of tumors induced by SV40 was consistently related with the development of this antibody. Adult hamsters vaccinated with SV40 and subsequently resisting a transplant of SV40 transformed cells developed similar antibody in their sera which could be detected as early as the 2nd week after the challenge. The evidence indicates that the S antibodies elicited by the techniques employed may be directed against SV40-specific transplantation antigens.