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Concordant segregation analysis of Chinese hamster (Cricetulus griseus) isozymes and chromosomes segregating from hamster X mouse interspecific somatic cell hybrids revealed that loci for ME1, PGM3, MPI, and PKM2 are located on Chinese hamster chromosome 4. Synteny of these loci in hamsters provides additional evidence for the conservation of mammalian autosomal linkage groups.
The laboratory mouse (Mus musculus, 2n = 40), the Chinese hamster (Cricetulus griseus, 2n = 22), and the golden (Syrian) hamster (Mesocricetus auratus, 2n = 44) are common laboratory animals, extensively used in biomedical research. In contrast with the mouse genome, which was sequenced and well characterized, the hamster species has been set aside. We constructed a chromosome paint set for the golden hamster, which for the first time allowed us to perform multidirectional chromosome painting between the golden hamster and the mouse and between the two species of hamster. From these data we constructed a detailed comparative chromosome map of the laboratory mouse and the two hamster species. The golden hamster painting probes revealed 25 autosomal segments in the Chinese hamster and 43 in the mouse. Using the Chinese hamster probes, 23 conserved segments were found in the golden hamster karyotype. The mouse probes revealed 42 conserved autosomal segments in the golden hamster karyotype. The two largest chromosomes of the Chinese hamster (1 and 2) are homologous to seven and five chromosomes of the golden hamster, respectively. The golden hamster karyotype can be transformed into the Chinese hamster karyotype by 15 fusions and 3 fissions. Previous reconstructions of the ancestral murid karyotype proposed diploid numbers from 2n = 52 to 2n = 54. By integrating the new multidirectional chromosome painting data presented here with previous comparative genomics data, we can propose that syntenies to mouse Chrs 6 and 16 were both present and to hypothesize a diploid number of 2n = 48 for the ancestral Murinae/Cricetinae karyotype.
We studied the clastogenic action of cyclophosphamide (CP) on bone marrow cells of the Armenian hamster (AH), Cricetulus migratorius. CP induced a dose-dependent linear increase in aberrant cells. The maximal cytogenetic action was observed 12 h after CP treatment. Male and female AHs were similarly sensitive to the clastogenic action of CP. We compared CP clastogenicity at a dose of 25 mg/kg on bone marrow cells of AHs, mice, rats, guinea pigs and Chinese hamsters 24 h after treatment. We observed that this dose of CP induced only 2.8% aberrant cells in bone marrow of AHs, but 42.8%, 32.2%, 25% and 14.6% aberrant cells in bone marrow of guinea pigs, rats, mice and Chinese hamsters respectively. AHs are much more resistant to the metaphase-arresting action of colchicine than other species of rodents (e.g., the colchicine dose for AHs is 100-fold more than for rats). Thus AHs are the most resistant of all rodent species studied to the clastogenic action of CP.
Thiotepa (TT) induced chromosomal aberrations in the bone marrow cells of the Armenian hamster, Cricetulus migratorius, in a dose-dependent manner. The maximal clastogenic action was observed 24 h after treatment with thiotepa. Male and female Armenian hamsters were similarly sensitive to the clastogenic action of thiotepa. The lowest effective concentration of thiotepa for Armenian hamsters was 2.5 mg/kg b.w. (6.1% of the LD50). The results obtained and analysis of the literature data have shown that Armenian hamsters are much more resistant to the clastogenic action of thiotepa than various other species of rodent. This resistance is not due to the metabolic activation rates of chemical mutagen in the liver of Armenian hamsters, since thiotepa is a direct-acting clastogen.
The frequency of chromosomal aberrations in bone marrow cells, after a single i.p. aflatoxin B1 (AFB1) dose, was examined in male Chinese hamsters (Cricetulus griseus). There was a significant increase in aberrant cells within 5 days of administration of a dose of 0.1 micrograms-5 mg AFB1/kg, and on the 36th day. After a single dose of 5 mg AFB1/kg the enhanced frequency of aberrant cells was monitored up to day 104 with no sign of a decrease to control level. The results indicate that the minimum mutagenic effect of an AFB1 dose in this system is 0.1 micrograms/kg. Attention is drawn to the long-term presence of chromosomal aberrations even after a single i.p. exposure to AFB1.
Chinese hamsters (Cricetulus griseus) were treated with ethanol, with cigarette smoke and with both. During the experimental period of 12 weeks a control group of animals (c) received water ad libitum, another water drinking group received a cigarette smoke treatment during the last 4 weeks (S). Another group received 20% (v/v) ethanol during the whole experimental period as the only liquid supply (E), and one group with the same ethanol treatment was simultaneously treated with cigarette smoke during the last 4 weeks of the experiment (ES). The investigation of bone marrow cells after 12 weeks with regard to chromosomal aberrations and sister chromatid exchanges revealed no effects. A high mitotic activity was found in the smoke treated groups.
1. Three major DNA repetition classes can be distinguished in the genomes of three hamster species Mesocricetus auratus, Cricetulus griseus, and Phodopus sungorus sungorus: A very fast reassociating fraction of about 13% of the DNA, a fast reassociating fraction comprising 10-23% of the DNA, and a slowly reassociating fraction containing single-copy sequences of 63-78% of hamster DNA. 2. In the DNA of the Syrian and Djungarian hamster 43%-53%, and in the DNA of the Chinese hamster 80% of the single-copy sequences are interspersed with repetitive sequences. 3. The lengths of repetitive DNA sequences, vary between more than 3 kb and 0.3 kb, with the majority of sequences having a length of 0.3 kb to 0.5 kb. 4. The findings suggest that the genomes of the hamster species studied are organized in a short period interspersion pattern. 5. Unlike other vertebrate and invertebrate genomes, moderately repetitive sequences exhibit a high degree of intraspecific homology in the hamster genome.
Mitochondrial DNA from heart, liver and kidney of two hamster species, Mesocricetus auratus and Cricetulus griseus has been digested with the restriction endonucleases Bam HI, Bgl I, Eco RI, Hae III, Hind III, Hpa II and Xba I. Cleavage patterns for Hpa II are identical for both species, while only two of seven bands are common with Hae III. All other endonucleases give a species-specific cleavage pattern. The results suggest a fairly high phylogenetic differentiation of the mitochondrial genome between the two hamster species. The large differences in mitochondrial DNA variability between different species is discussed as a function of mutation rate and species-specific generation time.
Styrene produced no effect on induction of sex-chromosome nondisjunction in Drosophila melanogaster fed on 500 ppm styrene for 24 h. No increase in the frequency of micronuclei from chromosome fragments or from nondisjunction of whole chromosomes was seen in the bone marrow of Chinese hamsters (Cricetulus griseus) injected (i.p.) with a single dose of 1.0 g/kg b.wt. styrene or with 250 mg/kg b.wt. of the primary metabolite, styrene oxide.
Chinese hamster ovary cells have been widely used to manufacture recombinant proteins for human therapeutic use. A sensitive quantitative real-time polymerase chain reaction assay for the detection of residual Chinese hamster (Cricetulus griseus) DNA is presented in this paper. The assay is reasonably affordable and can be adapted for high-throughput screening using 96-well format. Real-time PCR primers were designed to amplify a 150bp region of a genomic fragment from hamster DNA. The specificity of the probe was evaluated in real-time PCR reactions using genomic DNA from mouse fibroblast, human kidney and hamster ovary cell lines as template. Sensitivity of real-time PCR was compared on genomic DNA from hamster cell line CHO DG44. These primers can be used in real-time PCR reactions to detect presence of contaminating hamster DNA in purified protein samples down to sensitivity of 300fg genomic DNA.
This study examined the behavioral and physiological effects of long-term exposure to overdose of aversive odor (predator odor) in two species of hamsters. About 0.05 mg of anal gland secretions of Siberian weasels (Mustela sibirica) was smeared at the oronasal groove of wild male ratlike hamsters (Cricetulus triton) (natural prey) and laboratory golden hamsters (Mesocricetus auratus) once every day for 4 weeks. After 28 days, the experimental groups of both hamster species displayed higher cortisol level, larger adrenal gland (in ratlike hamsters only), smaller thymus and flank gland, and lower aggression level than the conspecific control group (presented with water). Thus, the long-term presence of overdose of the anal gland secretion of the Siberian weasel could lower the aggression and social rank and suppress the immunity in the hamsters. The reproductive conditions of these prey species, however, seemed not to be affected. In addition, the similarities in the behavioral and physiological responses to the predator odor between the two species of hamsters showed that the responses to predator odor might be innate.
To enhance potential use of the Chinese hamster, Cricetulus griseus, in developmental and cytogenetic studies of mammalian gametes and embryos, techniques for in vitro fertilisation and embryo culture were developed in the species. Spermatozoa were recovered from the vasa deferentia of mature males, and incubated in modified TYH medium for 1 h at 37 degrees C under 5% CO2 in air. They were then treated with ionophore A23187 (20 microM) for 10 min to induce the acrosome reaction. Following ionophore treatment, superovulated oocytes were collected from hormonally stimulated females and incubated with the acrosome-reacted spermatozoa for 2 h at 37 degrees C under 5% CO2 in air. In this study, 245 oocytes were used for insemination, and 198 (80.8%) were found to be penetrated by sperm; among them, 194 ova (98.0%) were determined to be monospermic. The monospermic ova were then cultured in TYH supplemented with 1 mM hypotaurine under the same gas phase. Within 30 h of fertilisation, 182 ova (93.8%) cleaved to the 2-cell stage, and subsequently 163 ova (84.0%) developed beyond the 2-cell stage. Thus, obstinate developmental arrest at the 2-cell stage ('2-cell block') was not observed in this species. Ultimately, 65.5% of monospermic ova reached morula to blastocyst stages.
Analysis of aminoacylase-1 (ACY-1) zymograms from 21 species indicated one structural locus coding for one dimeric enzyme. Among the various strains of five laboratory mammals (Mesocricetus auratus, Cricetulus griseus, Mus musculus, Rattus norvegicus, Cavia porcellus) only in Mus musculus-strains genetic variability was found. Crosses segregating for ACY-1b/ACY-1c reveal a three-banded banding pattern typical for the heterozygous state of a dimeric enzyme. Electrophoretic comparisons of all species studied indicate a high degree of phylogenetic variability at this locus.
Elk3 belongs to the Ets family of transcription factors, which are regulated by the Ras/mitogen-activated protein kinase-signaling pathway. In the absence of Ras, this protein is a strong inhibitor of transcription and may be directly involved in regulation of growth by downregulating the transcription of genes that are activated during entry into G1. We have isolated the Cricetulus griseus Elk3 gene from the Chinese hamster ovary (CHO) cell line and investigated the transcriptional potential of this factor. Transient transfections revealed that, in addition to its regulation of the c-fos promoter, Elk3 from CHO cells seems to inhibit other promoters controlling expression of proteins involved in G1/S phase progression; Cyclin D1 and DHFR. As has been described for the Elk3 homologs Net (Mouse) and Sap-2 (Human), the results of the present study further indicate that hamster Elk3 is a target of the Ras-Raf-MAPK pathway, and cotransfections with constitutively active H-ras relieves its negative transcriptional activity. No cells stably expressing exogenous Elk3 could be obtained, possibly due to an unspecified toxic or growth retarding effect. These findings support a possible role for Elk3 in growth regulation and reveal a high degree of homology for this protein across species.
We describe the construction and analysis of recombinant DNA libraries representative of chromosomes 1 and 2 of Chinese hamster (Cricetulus griseus). Propidium-iodide stained chromosomes were purified by flow cytometric analysis and sorting, and EcoRI digests of purified DNA were cloned into the bacteriophage vector Charon 4A. These libraries contain DNA complementary to 63% and 69% of nick-translated DNA derived from flow-purified chromosomes 1 and 2, respectively. However, sequences complementary to only 24% and 35% of a total Chinese hamster genomic DNA tracer were hybridized in parallel renaturation experiments. The chromosome 2 library contained DNA sequences encoding dihydrofolate reductase (dhfr), a gene previously mapped to Chinese hamster chromosome 2. No sequences complementary to dhfr were found in the library constructed from chromosome 1 DNA. These analyses are discussed with regard to the current limitations and future strategies for the construction of chromosome-specific DNA sequence libraries of high purity and completeness.
Careful analysis of G-band patterns in various rodent families allows identification of homology and thus accurate prediction of gene map positions. However, conclusions based on the synteny of genes without a careful study of chromosome evolution and G-band homology can be misleading. We tested these generalizations by means of G-band analysis and in situ hybridization with three genes in Chinese hamster (Cricetulus griseus, CGR) chromosomes. The location of the adenosine deaminase gene, previously mapped by somatic cell hybrid panels, was confirmed and further sublocalized on CGR 6q1. Although transferrin and uridine monophosphate synthetase are localized to adjacent bands on human chromosome 3 (3q21 and 3q13, respectively), we report that these genes are widely separated on CGR 4q2 and 4p2, respectively.
The serum of Armenian hamster (Cricetulus migratorius) contains a protein homologous to female protein (FP) that has been characterized in the Syrian (golden) hamster. Of unknown function, FP belongs to a family of proteins (called pentraxins) that have a common ancestral gene and are widely expressed in nature. Whereas serum concentration of FP in Syrian hamsters (SFP) is many fold greater (200- to 300-fold) in females vs. males, Armenian hamster FP (AFP) is only moderately elevated (approximately 3-fold) in female vs. males and only for the fall-winter months of the year. In the Armenian hamster testosterone administration to females or castration of males has no effect on AFP serum levels, whereas in Syrian hamster these treatments change SFP serum concentration to that characteristic of the opposite sex. Some sex steroid control of hepatic AFP synthesis is evident, however, as serum levels decrease after exogenous estrogen treatment. In contrast to Syrian hamster FP, normal levels of AFP are dependent on an intact hypophysial-pituitary axis and also are influenced by the season of the year. As an acute-phase protein, AFP responds in a typical fashion, with increasing serum levels detected in both sexes in contrast to the divergent sex-limited response in Syrian hamsters. Although AFP and SFP are similar structurally, morphologically, and antigenetically and share common binding specificities, the regulation of FP synthesis in Armenian hamster is very different from that previously found in Syrian hamster.
Following testicular X-irradiation of Chinese hamster (Cricetulus griseus) males eight progeny (five males and three females) with reciprocal translocatons (heterozygotes) were obtained. They were all phenotypically normal but semi-sterile. Homozygotes were produced successfully with CHAT3 (2p-;9q+) and CHAT5 (2q-;8q+). They were proven to be fertile. Thus, two homozygous stocks with distinct marker chromosomes were established. CHAT3 homozygotes have shown a high incidence of diabetes, while CHAT5 nomozygotes had no genetic burden. Progeny tests of outcrosses and intercrosses with the heterozygotes and homozygotes were performed in both stocks. Karyological analysis of the first cleavage division showed the marker chromosomes of both stocks to be useful in distinguishing the paternal chromosome complements from the maternal ones.