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PubMed2007

Somatic mutations at the trinucleotide repeats of androgen receptor gene in male hepatocellular carcinoma.

Androgen and androgen receptor (AR) have long been implicated in liver carcinogenesis, especially for the male dominance feature. However, whether AR gene could occur in somatic mutations that might contribute to this process has not yet been studied. DNA sequencing and genotyping were conducted for detecting the genetic aberrations of AR gene in 257 primary hepatocellular carcinomas (HCCs) and also the dysplastic nodules (DN) from another 11 patients. Twenty-one AR somatic mutations causing amino acid changes were identified in HCC and even in the precancerous DN. The missense somatic mutations of AR were rare in HCC (2 cases) but the trinucleotide repeat (TNR) changes, both at (CAG)n and (GGC)n, was a more common one (19 cases). Notably, all these mutations occurred in male patients and most TNR changes belonged to the contraction type (15 out of 19 cases, 78.9%), which has been reported to associate with increased AR transcriptional activity. Most samples with TNR changes did not show microsatellite instability, suggesting a different cause for these TNR mutations. Although no significant correlation was identified between AR mutations and the clinicopathologic parameters, we found the (CAG)n length significantly shorter in hepatitis B virus (HBV)(+) HCCs than in HBV(-) HCCs and the (GGC)n length significantly correlates with the overall survival. In conclusion, the mis-sense somatic mutations of AR were rare in HCC but the TNR change was a more common one, which exclusively occurred in males. Moreover, the length of TNR carried clinical significance in special HCC group.

Adult↗

PubMed1995

Constitutional and somatic mutations in the WT1 gene in Wilms' tumor patients.

Wilms' tumor development, like most tumors, involves multiple genetic alterations affecting diverse genes. Only one of these has thus far been identified, the Wilms' tumor 1 (WT1) gene on 11p13, which functions as a tumor suppressor gene. We assessed the involvement of the WT1 gene constitutionally and somatically in 26 Wilms' tumor patients. Of these, the clinical data suggest a constitutional pre-disposition in 12 bilateral cases and 7 cases with early onset. We employed Southern blot analysis and PCR-based markers for analyses of somatic allelic losses in chromosome bands 11p13, 11p15 and 16q and screened for point mutations in exons 2-10 of the WT1 gene with denaturing gradient gel electrophoresis (DGGE). Of the 12 cases with multiple tumors, 1 exhibited a constitutional 11p13 deletion and a somatic stop mutation in exon 4 of the WT1 gene and 2 harbored constitutional mutations in the WT1 gene: a pre-mature stop codon in exon 6 in a boy with bilateral cryptorchidism and bilateral Wilms' tumors and an intragenic deletion in a girl with bilateral WT. Three additional bilateral tumors displayed WT1 rearrangements or allelic losses with 11p13 markers. Four of 7 patients with an early onset of unilateral tumor had losses of 11p13, though no WT1 mutations were detected. Two of the remaining cases that did not show any somatic or constitutional 11p13 alterations had Beckwith-Wiedemann syndrome, known to involve the 11p15 region.

Alleles↗

PubMed1987

A new lamin in Xenopus somatic tissues displays strong homology to human lamin A.

The nuclear lamina of vertebrates is composed of several major polypeptides that range in mol. wt from 60 to 80 kd. In mammals, the three major lamin proteins are designated A, B and C. Two major lamins have been described in Xenopus somatic tissues; two other lamins are expressed primarily in germ cells. We have analysed a cDNA clone encoding a Xenopus lamin that is highly homologous to human lamins A and C. The predicted protein has the carboxy-terminal domain characteristic of human lamin A and is thus a lamin A homologue. Surprisingly, the lamin encoded by the cDNA clone is not one of the known Xenopus lamins. The encoded protein is distinct in size from the oocyte lamin LIII and the two somatic lamins LI and LII. Monoclonal antibodies specific for LII, LIII and LIV (the lamin of male germ cells) do not recognize the protein encoded by the cDNA clone; conversely, a polyclonal antibody against the encoded protein does not recognize any of the known Xenopus lamins. This lamin is expressed late in embryonic development, and is present in all adult somatic cells examined, except erythrocytes. Thus frogs and mammals are similar in having three major somatic lamins that fall into distinct structural classes.

Animals↗

PubMed1982

Comparison of somatic and germ cell models for cytogenetic screening.

A variety of in vivo mammalian test models are available for screening of chemicals for mutagenicity at the chromosomal level. These models have been grouped into those focusing on somatic cell effects and those dealing with germ cell effects. An analysis of available literature indicates that 76 compounds have been tested from chromosome effects in both somatic and germ cells. Of these, concordant results (positive-positive or negative-negative) were obtained with 58 compounds. Of the remaining 18 compounds with discordant results, all were positive in somatic cells, but negative in germ cell assays. These results suggest an inherent relative insensitivity of germ cells themselves to mutagenic chemicals. In the context of screening for safety evaluation purposes, this analysis suggests that a negative somatic-cell response can be taken as highly predictive of negative results in a germ cell assessment.

Animals↗

PubMed2007

The radiation-induced block in spermatogonial differentiation is due to damage to the somatic environment, not the germ cells.

Radiation and chemotherapeutic drugs cause permanent sterility in male rats, not by killing most of the spermatogonial stem cells, but by blocking their differentiation in a testosterone-dependent manner. However, it is not known whether radiation induces this block by altering the germ or the somatic cells. To address this question, we transplanted populations of rat testicular cells containing stem spermatogonia and expressing the green fluorescent protein (GFP) transgene into various hosts. Transplantation of the stem spermatogonia from irradiated adult rats into the testes of irradiated nude mice, which do not show the differentiation block of their own spermatogonia, permitted differentiation of the rat spermatogonia into spermatozoa. Conversely transplantation of spermatogonial stem cells from untreated prepubertal rats into irradiated rat testes showed that the donor spermatogonia were able to colonize along the basement membrane of the seminiferous tubules but could not differentiate. Finally, suppression of testosterone in the recipient irradiated rats allowed the differentiation of the transplanted spermatogonia. These results conclusively show that the defect caused by radiation in the rat testes that results in the block of spermatogonial differentiation is due to injury to the somatic compartment. We also observed colonization of tubules by transplanted Sertoli cells from immature rats. The present results suggest that transplantation of spermatogonia, harvested from prepubertal testes to adult testes that have been exposed to cytotoxic therapy might be limited by the somatic damage and may require hormonal treatments or transplantation of somatic elements to restore the ability of the tissue to support spermatogenesis.

Animals↗

PubMed2006

Anxiety sensitivity and depression: mechanisms for understanding somatic complaints in veterans with posttraumatic stress disorder.

A study was conducted among 45 male veterans seeking inpatient treatment for posttraumatic stress disorder (PTSD) to test whether the relationship between PTSD and somatic complaints was accounted for by depression and anxiety sensitivity. Posttraumatic stress disorder symptom severity, depression symptom severity, and anxiety sensitivity were each positively and significantly related to veterans' self-reported severity of somatic complaints. Results of hierarchical regression analyses indicated that anxiety sensitivity and depression severity account for the relationship between PTSD and veterans' somatic complaints, suggesting PTSD influences somatic complaints by virtue of underlying symptoms of depression and anxiety sensitivity.

Anxiety↗

PubMed2002

Estimating the prevalence of mental and somatic disorders in the community: aims and methods of the German National Health Interview and Examination Survey.

This paper outlines the principal aims and design of the German National Health Interview and Examination Survey, mental health supplement (GHS-MHS), the first nationwide, epidemiological study of both somatic and mental health in Germany on a representative sample of 4,181 subjects in the community. Both the broader context of the study--in particular its methodological relation to the social and somatic core survey of the German National Health Interview and Examination Survey (GHS-CS)--and the internal methodology of the mental health supplement (GHS-MHS) are presented. The study's strategies and method are derived from a consideration of important theoretical issues arising from epidemiological studies in the field of public health. The main instrument used to assess diagnoses of mental disorders was a standardized diagnostic interview for mental disorders (following DSM-IV (CIDI)) applied by clinically trained interviewers. This diagnostic interview was supplemented by modules on comorbidity, help seeking, treatment and impairment. Somatic health diagnoses were made using an integrated approach including self-report measures, a standardized clinical interview, and laboratory measures. Findings on sampling, response rate, weighting and sample characteristics are presented. Critical issues are discussed, including the scientific objectives that have been achieved by the study. Overall, the GHS core survey and its mental health supplement provide the mental health research community with complex data that allow for high-quality analysis of mental disorders and associations with somatic disorders.

Adolescent↗

PubMed2002

Differential cytoplast requirement for embryonic and somatic cell nuclear transfer in cattle.

Effective activation of a recipient oocyte and its compatibility with the nuclear donor are critical to the successful nuclear reprogramming during nuclear transfer. We designed a series of experiments using various activation methods to determine the optimum activation efficiency of bovine oocytes. We then performed nuclear transfer (NT) of embryonic and somatic cells into cytoplasts presumably at G1/S phase (with prior activation) or at metaphase II (MII, without prior activation). Oocytes at 24 hr of maturation in vitro were activated with various combinations of calcium ionophore A23187 (A187) (5 microM, 5 min), electric pulse (EP), ethanol (7%, 7 min), cycloheximide (CHX) (10 micro g/ml, 6 hr), and then cultured in cytochalasin D (CD) for a total of 18 hr. Through a series of experiments (Exp. 1-4), an improved activation protocol (A187/EP/CHX/CD) was identified and used for comparison of NT efficiency of embryonic versus somatic donor cells (Exp. 5). When embryonic cells from morula and blastocysts (BL) were used as nuclear donors, a significantly higher rate of blastocyst development from cloned embryos was obtained with G1/S phase cytoplasts than with MII-phase cytoplasts (36 vs. 11%, P < 0.05). In contrast, when skin fibroblasts were used as donor cells, the use of an MII cytoplast (vs. G1/S phase) was imperative for blastocyst development (30 vs. 6%, P < 0.05). Differential staining showed that parthenogenetic, embryonic, and somatic cloned BL contained 26, 29, and 33% presumptive inner cell mass (ICM) cells, respectively, which is similar to that of frozen-thawed in vivo embryos at a comparable developmental stage (23%). These data indicate that embryonic and somatic nuclei require different recipient cytoplast environment for remodeling/ reprogramming, and this is likely due to the different cell cycle stage and profiles of molecular differentiation of the transferred donor nuclei.

Animals↗

PubMed2001

Novel growth factor supporting survival of murine primordial germ cells: evidence from conditioned medium of ter fetal gonadal somatic cells.

The ter (teratoma, chromosome 18) mutation causes a deficiency of primordial germ cells (PGCs) in ter/ter embryos from the ter congenic mouse strain at 8.0 days post coitum (dpc). In order to analyse the function of the ter gene, here we examined effects of conditioned medium (CM) from 14.5 dpc testicular and ovarian somatic cells of +/+, +/ter, or ter/ter genotype on mouse PGCs "mixed-cultured" with own somatic cells on feeder cells. The results showed that +/+ and +/ter CM supported survival in 9.5 and 11.5 dpc ICR PGCs but ter/ter CM did not rescue TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling)-positive apoptosis in the PGCs though it did not affect 5-bromo-2-deoxyuridine incorporation in PGCs. This supportive substance in +/+ CM, not ter/ter CM, was characterized as soluble, heat labile, and larger than 30 kDa. We also found that several known growth factors for PGCs and their receptors were expressed in ter/ter testes as well as +/+ testes, suggesting the ter function is independent. Thus, it was concluded that fetal gonadal somatic cells express a novel PGC growth factor (designated as TER Factor) supporting survival of PGCs not somatic cells and that the PGC deficiency in ter/ter testes is caused by a loss of this factor.

Animals↗

PubMed2004

Incidence of apoptosis in clone embryos and improved development by the treatment of donor somatic cells with putative apoptosis inhibitors.

This study was conducted to promote in vitro-development of clone embryos by the treatment of donor somatic cells with hemoglobin (Hb) and/or beta-mercaptoethanol (ME), based on the analysis of apoptosis after somatic cell nuclear transfer (SCNT). Prospective, randomized study was conducted and, in vitro-matured bovine oocytes and fetal fibroblasts were provided for SCNT. In the first series of experiment, embryo apoptosis after SCNT was monitored by a terminal deoxynucleotidyl transferase-mediated d-UTP nick end-labeling assay. As results, apoptosis occurred more (P < 0.05) frequently after SCNT than after in vitro-fertilization (IVF) of control treatment. Subsequently, donor somatic cells treated with Hb (1 microg/ml) and/or ME (10 microM) were provided for SCNT. Either Hb or ME greatly reduced apoptosis (0.083 +/- 0.006 vs. 0.058-0.068 +/- 0.005), while combined treatment did not. ME was more promotive than Hb; significant increases were found in morula compaction (86%), cell numbers of blastocyst (131.3 +/- 1.3 cells/blastocyst), and inner cell mass (31.9 +/- 0.8 cells/blastocyst) cell, and the ratio of inner cell mass to trophectodermal cell numbers (0.24 +/- 0.01). In conclusion, the treatment of donor somatic cells with ME or Hb could reduce apoptosis after SCNT, resulting improved preimplantation development.

Animals↗

PubMed2006

Rat Spag5 associates in somatic cells with endoplasmic reticulum and microtubules but in spermatozoa with outer dense fibers.

The leucine zipper motif has been identified as an important and specific interaction motif used by various sperm tail proteins that localize to the outer dense fibers. We had found that rat Odf1, a major integral ODF protein, utilizes its leucine zipper to associate with Odf2, another major ODF protein, Spag4 which localizes to the interface between ODF and axonemal microtubule doublets, and Spag5. The rat Spag5 sequence indicated a close relationship with human Astrin, a microtubule-binding spindle protein suggesting that Spag5, like Spag4, may associate with the sperm tail axoneme. RT PCR assays indicated expression of Spag5 in various tissues and in somatic cells Spag5 localizes to endoplasmic reticulum and microtubules, as expected for an Astrin orthologue. MT binding was confirmed both in vivo and in in vitro MT-binding assays: somatic cells contain a 58 kDa MT-associated Spag5 protein. Western blotting assays of rat somatic cells and male germ cells at different stages of development using anti-Spag5 antibodies demonstrated that the protein expression pattern changes during spermatogenesis and that sperm tails contain a 58 kDa Spag5 protein. Use of affinity-purified anti-Spag5 antibodies in immuno electron microscopy shows that in rat elongated spermatids and epididymal sperm the Spag5 protein associates with ODF, but not with the axonemal MTs. This observation is in contrast to that for the other Odf1-binding, MT-binding protein Spag4, which is present between ODF and axoneme. Our data demonstrate that Spag5 has different localization in somatic versus male germ cells suggesting the possibility of different function.

Animals↗

PubMed2007

Transfer of spermatogenesis-related cDNAs into eel testis germ-somatic cell coculture pellets by electroporation: methods for analysis of gene function.

Genes encoding spermatogenesis-related substance (eSRSs) show unique expression patterns during spermatogenesis. To analyze their function, we developed a new assay system using gene transfer techniques combined with coculture of the eel germ-somatic cells. First, we investigated the efficacy of in vitro electroporation transfer of gene into germ-somatic cell pellets using green fluorescent protein (GFP) gene. Second, in order to define the function of the eSRSs, we electrophoretically transferred eel spermatogonial stem cell renewal factor (eSRS34) and eel spermatogenesis-preventing substance (eSRS21) genes into germ-somatic cell pellets. Presence of the transferred cDNA was examined by reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, proliferating cells were detected histologically, after labeling with BrdU. Transfer of the eSRS34 gene induced spermatogonial stem cell renewal in the pellets. Moreover, 11-ketotestosterone (11-KT) treatment stimulated the proliferation of spermatogonia, which resulted in the appearance of late type B spermatogonia in the pellets. The proliferation of spermatogonia by 11-KT stimulation was suppressed by transfer of the eSRS21 gene. These results indicate that the transferred eSRS34 and 21genes were functional in the pellets. Thus, an efficient in vitro gene transfer technique for coculture system of germ and somatic cell of Japanese eel was established.

Anguilla↗

PubMed2006

Somatic awareness, uncertainty, and delay in care-seeking in acute heart failure.

Symptom monitoring is difficult for heart failure (HF) patients. Difficulties physically sensing and determining symptom meaning may lead to uncertainty and delay treatment. Somatic awareness may provide insight into symptom monitoring ability. A model combining physical and cognitive aspects of the symptom experience was developed to examine factors affecting care-seeking among HF patients. Adults hospitalized with acute HF were interviewed and completed questionnaires measuring somatic awareness and uncertainty. HF symptom duration prior to admission measured delay. HF specific somatic awareness and symptom pattern predicted delay. Uncertainty correlated with somatic awareness, but did not predict delay. Few responded to early HF symptoms delaying until acutely ill. Development of interventions to improve symptom monitoring is needed.

Acute Disease↗

PubMed1997

Soma-germ cell interactions in Caenorhabditis elegans: multiple events of hermaphrodite germline development require the somatic sheath and spermathecal lineages.

Germ cells complete multiple events to form functional oocytes and sperm. In the Caenorhabditis elegans hermaphrodite, germ cells develop in proximity to the somatic gonad sheath and spermathecal cells. We present evidence from cellular laser ablation studies indicating that cells of the somatic sheath and spermathecal lineages play critical roles in four events of hermaphrodite germline development. (1) Cells of the sheath and spermathecal lineage support germline proliferation; ablation of sheath/spermathecal precursor cells reduces mitotic proliferation. (2) These cells also play a role in the exit of germ cells from the pachytene stage of meiotic prophase and/or gamete differentiation; ablation can result in undifferentiated germ cells arrested in pachytene. (3) Proximal sheath and distal spermatheca cells are required for ovulation of the oocyte. During wild-type ovulation, the mature oocyte is expelled from the gonad arm by contraction of the proximal myoepithelial sheath and dilation of the distal spermatheca. Ablation of these cells traps mature oocytes in the gonad arm where they endomitotically replicate their DNA (the Emo phenotype). (4) Cells of the sheath and spermathecal lineage also appear to promote the male germ cell fate since ablation of one sheath/spermathecal precursor cell can feminize the hermaphrodite germ line. These somatic ablation-induced germline phenotypes demonstrate that the somatic gonad is required for multiple events in C. elegans germline development. Further, these results suggest that soma to germline cell-cell interactions in C. elegans are physiological in character (i.e., contraction during ovulation) as well as regulatory.

Animals↗

PubMed1997

Misexpression of nautilus induces myogenesis in cardioblasts and alters the pattern of somatic muscle fibers.

nautilus (nau), one member of the myogenic regulatory family of bHLH-encoding genes, is expressed in a subset of muscle precursors and differentiated fibers in the Drosophila embryo. To elucidate the role of nautilus in myogenesis, we have misexpressed it using the GAL4-targeted system. We find that ectopic expression results in lethality throughout Drosophila development. We analyzed the effects of embryonic expression in mesodermal tissues that include the cardioblasts of the dorsal vessel as well most, if not all, of the presumptive somatic muscle precursors. Immunohistochemical staining for muscle MHC revealed abnormalities that include an absence of cardial cells, coincident with the appearance of novel muscle fibers adjacent to the dorsal vessel. Moreover, many cardioblasts express increased levels of muscle-specific genes such as myosin, actin 57B, and Mlp60A, a protein that is restricted to the somatic, visceral, and pharyngeal muscles. These data suggest that the missing cardial cells have been transformed into cells with properties similar to those of the somatic muscles. In addition, ubiquitous expression of nautilus in somatic muscle cells of these embryos resulted in muscle pattern defects. Specifically, muscles that do not normally express nautilus were frequently absent, and novel fibers were observed in positions reminiscent of nau-expressing muscles. These data imply that nautilus can alter the developmental program of muscle precursors. In summary, we suggest that nautilus induces myogenic differentiation in vivo when ectopically expressed in developing cardioblasts and may affect the myogenic differentiation program of specific muscle fibers.

Animals↗

PubMed1999

Transcription of the myogenic regulatory gene Mef2 in cardiac, somatic, and visceral muscle cell lineages is regulated by a Tinman-dependent core enhancer.

The MADS-box transcription factor MEF2 is expressed specifically in developing cardiac, somatic, and visceral muscle cell lineages during Drosophila embryogenesis and is required for myoblast differentiation and muscle morphogenesis. To define the mechanisms that regulate Mef2 transcription, we have analyzed the Mef2 upstream region for sequences sufficient to recapitulate the expression pattern of the gene in Drosophila embryos. Here we describe a complex enhancer located 5.8 kb upstream of the Drosophila Mef2 gene that controls transcription in cardial cells of the dorsal vessel, a subset of somatic muscle founder cells, and the visceral muscle cells. The core of this enhancer contains two evolutionarily conserved binding sites for the homeodomain protein Tinman (Tin), expressed in developing cardiac, somatic, and visceral muscle lineages. Both Tin binding sites are required for enhancer activity in all three muscle cell lineages. Whereas the 285-bp enhancer core alone is sufficient for expression in cardiac cells, expression in somatic founder cells and visceral muscle is dependent on the core enhancer plus unique flanking sequences that include an evolutionarily conserved E box. These results reveal an essential role for Tin in activation of Mef2 transcription in multiple myogenic lineages and demonstrate that transcriptional activity of Tin is dependent on combinatorial interactions with other factors unique to different muscle cell types.

Animals↗

PubMed1996

Fusion with activated mouse oocytes modulates the transcriptional activity of introduced somatic cell nuclei.

We have analyzed the transcriptional activity of somatic cell nuclei fused with artificially activated mouse oocytes. Two types of somatic cells have been used: transcriptionally silent thymocytes, obtained from the thymus of newborn mice, and transcriptionally active murine erythroleukemia cells (MEL) from in vitro culture. Cells were fused with activated oocytes, either less than 1 h, or 3 h post-ethanol-treatment. When the fusion occurred early after activation (1 h or less), the transferred somatic cell nuclei reacted by nuclear envelope breakdown (NEBD), which exposed their chromatin to the cytoplasmic environment of the oocytes, and the reconstituted nuclei underwent significant ultrastructural remodeling. No transcriptional activity was ever detected in these reconstituted nuclei during the subsequent 3-4 h of culture of the resulting hybrid cells. In the case of MEL, this means that transcriptional activity ceased as soon as they entered the cytoplasm. In contrast, somatic nuclei which entered the cytoplasm of activated oocytes 3 h postactivation did not undergo NEBD and their remodeling was less pronounced. In contrast to the first group, these nuclei were transcriptionally active during the following 3-4 h of culture. In both cases, the female pronucleus remained transcriptionally silent.

Animals↗

PubMed1994

Somatic cell hybrid panel and NotI linking clones for physical mapping of human chromosome 3.

To identify by positional cloning the putative tumor-suppressor genes on the short arm of human chromosome 3 that are involved in the initiation or progression of several human malignancies, we have developed human mouse somatic cell hybrids and NotI linking libraries. The somatic cell hybrids contain either the intact human chromosome 3 or its derivatives as the only human genetic material in rodent background. The somatic cell hybrid panel defines five chromosomal regions on the short arm and two chromosomal regions on the long arm of chromosome 3. Two hundred sixty-one NotI linking probes from three independently constructed linking libraries were assigned to the seven chromosomal regions. The somatic cell hybrid panel and the regionally localized NotI linking probes should facilitate the construction of genetic linkage and physical maps to identify various tumor-suppressor and disease-related genes not only on the chromosome 3p, but on the entire chromosome.

Animals↗