Biomedical subjects

K Rhee

Publications and source records attributed to K Rhee.

At least 19 recordsLinked to original sources

PubMed2005

Effects of epidural naloxone on pruritus induced by epidural morphine: a randomized controlled trial.

BACKGROUND: Epidural morphine produces prolonged analgesia but has many side effects including pruritus. Naloxone is an antagonist that can reverse the side effects of morphine. METHOD: We studied the effects of continuously administered epidural naloxone mixed with morphine on side effects and analgesia in a randomized, double blind, two-armed study. Fifty-eight pregnant women undergoing cesarean section were enrolled. All patients received a 4-mg epidural bolus of morphine in the post-anesthetic care unit. After this, patients in group M (n=28) received continuous epidural morphine (6 mg over 48 h) in 0.1% bupivacaine; patients in group N (n=30) received an epidural infusion containing naloxone (1.2 mg over 48 h) and morphine (6 mg over 48 h) in 0.1% bupivacaine. The infusion rate was 2 mL/h. RESULTS: The incidence (82% versus 47%) and severity of pruritus were lower in group N than group M (P=0.001). There were no significant differences in pain score or in the incidence of nausea, vomiting or urinary disturbance between groups. CONCLUSION: Continuous epidural infusion of naloxone combined with morphine is effective in reducing the incidence and severity of pruritus induced by epidural morphine.

Adult↗

PubMed2003

Intravenous clonidine prolongs bupivacaine spinal anesthesia.

BACKGROUND: Prolongation of spinal anesthesia by oral clonidine premedication has been known. We hypothesized that intravenous clonidine administered after the spinal block may prolong spinal anesthesia. METHODS: To assess the prolongation of spinal anesthesia by intravenous clonidine, we designed a double-blind, placebo-controlled, prospective study. Patients scheduled for orthopedic surgery received 12 mg of 0.5% hyperbaric bupivacaine and were randomly divided into three groups (n = 26 in each group). In the clonidine 10-min group, 3 microg x kg-1 of clonidine was administered for 10 min immediately after the spinal block. In the clonidine 60-min group, 3 microg x kg-1 of clonidine was administered for 10 min, 50 min after the spinal block. The control group received normal saline. Sensory block was evaluated by pinprick and the duration was defined as the time for sensory block to regress to L1 dermatome. Duration of motor block was defined as the time required for the patient to flex his or her knee. RESULTS: The duration of sensory block was longer in both the clonidine 10-min and clonidine 60-min groups compared with the control group (196 +/- 42 min, 179 +/- 41 min vs. 125 +/- 25 min, P < 0.05). The duration of motor block was longer in the clonidine 10-min group than in the control group (153 +/- 26 min vs. 131 +/- 29 min, P < 0.05). The lowest heart rate and mean blood pressure were not different among groups. CONCLUSIONS: Intravenous clonidine administration within 1 h after the spinal block prolonged bupivacaine spinal anesthesia for approximately 1 h without adverse effects.

Adult↗

PubMed2001

9-cis-Retinoic acid represses transcription of the gonadotropin-releasing hormone (GnRH) gene via proximal promoter region that is distinct from all-trans-retinoic acid response element.

We previously reported an enhancing effect of all-trans-retinoic acid (all-trans-RA) on gonadotropin-releasing hormone (GnRH) gene transcription via distal promoter elements of the rat GnRH gene. The present study examined the effects of another biologically active retinoid, 9-cis-retinoic acid (9-cis-RA), on GnRH transcription in GT1-1 cells. Similar to the action of all-trans-RA, 9-cis-RA significantly induced the luciferase activity of the strong retinoic acid response element (RARE) reporter construct, 3X beta RARE-Luc, by about 60-fold, indicating that GT1-1 cells are also responsive to 9-cis-RA. In contrast to the stimulatory effect of all-trans-RA on GnRH transcription, 9-cis-RA inhibited the GnRH promoter activity in a dose- and time-dependent manner. Significant inhibition by 9-cis-RA required at least an 18 h treatment and a further decrease of GnRH promoter-driven luciferase activity was observed up to 48 h of incubation. Accordingly, GnRH mRNA levels were decreased by 9-cis-RA treatment in a similar dose- and time-related manner, indicating that mouse GnRH expression is also negatively regulated by 9-cis-RA. Transient transfections of serial deletion constructs of the rat GnRH promoter revealed that the --230/--110 sequence of the rat GnRH promoter is responsible for 9-cis-RA-induced inhibition of GnRH transcription. Within this region, however, no consensus retinoid X receptor response element was found. To gain insights into the role of retinoid X receptors (RXRs) in GnRH expression, we examined the effects of RXR overexpression on GnRH transcriptional activity. Interestingly, co-transfection of RXR overexpression vectors significantly increased the GnRH promoter-driven luciferase activity, while treatment with 9-cis-RA not only nullified the enhancing effect of RXR overexpression but also decreased the basal GnRH promoter-driven luciferase activity by 50% compared to vehicle-treated controls. This implies that RXRs in the absence of its cognate ligand 9-cis-RA contribute to the maintenance of basal GnRH gene transcription. Northern blot analysis revealed that 9-cis-RA, but not all-trans-RA, down-regulated RXR beta expression in GT1-1 cells, suggesting that one possible mechanism of 9-cis-RA-induced repression involves down-regulation of RXR expression. In conclusion, the present study clearly demonstrates that 9-cis-RA is a negative regulator of GnRH gene expression in immortalized GnRH neurons.

Alitretinoin↗

PubMed2001

A functional retinoic acid response element (RARE) is present within the distal promoter of the rat gonadotropin-releasing hormone (GnRH) gene.

We previously demonstrated that all-trans-retinoic acid (all-trans-RA) regulates gonadotropin-releasing hormone (GnRH) release and gene expression in rat hypothalamic fragments and GT1-1 neuronal cells. Promoter analysis of rat GnRH gene revealed that the enhancing effect of all-trans-RA on GnRH transcription is mediated by cis-elements localized within --1640/--1438 of the rat GnRH promoter. In the present study, we attempted to localize functional retinoic acid response elements (RAREs) within the all-trans-RA-responsive region of the rat GnRH gene. Sequence analysis showed that there exist three putative repeats of AGGTCA-related sequences (--1637/--1617, --1579/--1562, and --1494/--1470) within this promoter sequence. Among them, only the --1494/--1470 sequence could compete the specific binding of GT1-1 nuclear extracts to the consensus RARE (direct repeat of AGGTCA with a 5-bp spacer, DR-5) and vice versa in electrophoretic mobility shift assays. In addition, like consensus RARE, the --1494/--1470 sequence could confer all-trans-RA responsiveness when inserted into the upstream region of SV40 promoter. Treatment of GT1-1 cells with all-trans- or 9-cis-RA increased the specific bindings of GT1-1 nuclear extracts to the consensus RARE and to the --1494/--1470 sequence while not affecting the specific binding to the cAMP response element (CRE). Both retinoids induced RARbeta gene expression in GT1-1 cells. The --1494/--1470 sequence (5'-TCTTAGGACTCTGTGTGACCTAAGA) is similar to the direct repeat of TGACCT (complementary sequence of AGGTCA) with a spacer of 5 bp (i.e. DR-5 in the reverse orientation). A mutation of the second core recognition motif of the --1494/--1470 sequence to a more divergent one from consensus RARE (from TGACCT to TTACAT) abolished the responsiveness to all-trans-RA, whereas a mutation of first core recognition motif to a more TGACCT-like sequence (from AGGACT to TGAACT) increased the responsiveness to all-trans-RA. These results indicate that the --1494/--1470 sequence is indeed a weak but functional RARE of the modified DR-5 type. Taken together, these data indicate that all-trans-RA enhances GnRH transcription via functional RARE present in the distal region of the GnRH promoter.

Animals↗

PubMed2001

Heat stress attenuates ventilator-induced lung dysfunction in an ex vivo rat lung model.

Our laboratory has previously shown decreased mortality rates and the attenuation of lung injury in rats exposed to heat stress (H) 18 h prior to induction of sepsis. In the present study, we examined the hypothesis that heat stress would protect lungs against ventilator-induced lung injury. Male Sprague-Dawley rats were anesthetized and randomly allocated to receive either sham treatment or exposure to heat (rectal temperature 41 degrees C, for 15 min). The lungs were harvested 18 h later, a pressure-volume (P- V) curve was constructed, and the lungs were either lavaged for cytokine and surfactant analyses (preventilation data) or were mechanically ventilated with VT 40 ml/kg in a warmed, humidified chamber. After 2 h of mechanical ventilation, another P-V curve was constructed and the lungs were lavaged for cytokine and surfactant analyses (postventilation data). Mechanical ventilation in control lungs produced a 47% decrease in chord compliance, an increase in lung lavage levels of tumor necrosis factor (TNF)-alpha (722 +/- 306 pg/ml), interleukin (IL)-1beta (902 +/- 322 pg/ml), and macrophage inflammatory protein-2 (MIP-2) (363 +/- 104 pg/ml) as compared with low levels of cytokines detected in preventilation data, and no change in percentage of surfactant large aggregates (LA). In contrast, in mechanically ventilated lungs from animals that were exposed to heat stress we observed a smaller decrease in chord compliance (17%), a significant attenuation in cytokine levels (TNF-alpha 233 +/- 119 pg/ml; IL-1beta 124 +/- 53 pg/ml; MIP-2 73 +/- 52 pg/ml; p < 0.05) and a significant increase in percentage LA compared with control animals. We conclude that exposing animals to heat stress confers protection against the effects of an injurious form of mechanical ventilation, by a mechanism that may involve attenuation of cytokines and preservation of some surfactant properties.

Animals↗

PubMed2001

Marked tachypnea in siblings with chronic beryllium disease due to copper-beryllium alloy.

Two biological sisters working at the same factory for > 9 years developed chronic beryllium disease (CBD) from a copper-beryllium alloy. Both had marked tachypnea (36 breaths/min and 45 breaths/min at rest, respectively), persisting over 8 years. Ventilation during exercise (assessed in one sibling) was grossly excessive (57 L/min, expected 23 L/min) with a respiratory rate of 64 breaths/min even with normal baseline routine pulmonary function tests. Blood beryllium lymphocyte transformation test and lung biopsies confirmed the diagnosis. No other cases of CBD have been reported from this plant among about 120 workers. These cases support the genetic basis for berylliosis and illustrate the marked tachypnea that may accompany this disease.

Alloys↗

PubMed2001

Comparison of visual field defects between primary open-angle glaucoma and chronic primary angle-closure glaucoma in the early or moderate stage of the disease.

The visual field data (Humphrey C24-2) of early to moderate stage glaucoma between primary open-angle glaucoma (POAG) and chronic primary angle-closure glaucoma (PACG) patients was compared. Eleven POAG (11 eyes) and 14 chronic PACG patients (14 eyes) were matched with respect to age, sex, and disease severity. Although the mean deviation, pattern standard deviation, and short-term fluctuation were similar between the two groups, the mean corrected pattern standard deviation was higher in the POAG patients (p=0.048). When the numeric pattern deviation data (STATPAC II) was used for point-wise between-group comparisons, more depressed paracentral points were found in POAG patients. Different patterns were identified in visual field damages between the chronic PACG and POAG patients. The chronic PACG patients showed more generalized field loss than the POAG patients.

Aged↗

PubMed2001

Gray scale sonography of breast masses in adolescent girls.

OBJECTIVE: To assess the sonographic findings of breast masses in adolescents and the usefulness of sonographic patterns for suggesting a specific diagnosis. METHODS: The sonograms and medical records of 57 girls (mean age, 15.4 years) with palpable breast masses were retrospectively reviewed. Three observers reviewed the sonograms for multiple sonographic findings. Surgery or clinical findings established diagnoses. Statistical analysis was done to determine how well sonographic findings alone and in combination agreed with final histologic diagnoses. RESULTS: Diagnoses included cysts (n = 12), abscesses (n = 7), fibroadenomas (n = 36), a lactating adenoma (n = 1), and a phyllodes tumor (n = 1). The sonographic findings varied significantly among lesion types (P < or = .005). CONCLUSIONS: Our experience suggests that virtually all breast masses in a pediatric population are benign and that sonography has the ability to differentiate among cystic, inflammatory, and solid masses.

Abscess↗

PubMed1999

The cellular distribution and kinase activity of the Cdk family member Pctaire1 in the adult mouse brain and testis suggest functions in differentiation.

Pctaire1, a member of the family of cyclin-dependent kinases, has been shown to be particularly abundantly expressed in differentiated tissues such as testis and brain. However, very little is known about the cellular and subcellular distribution and function of Pctaire1 protein(s), which is the focus of this study. We show that Pctaire1 encoded two major proteins of M(r) approximately 62,000 and approximately 68,000, found predominantly in testis and brain. Within these two tissues, Pctaire1 was most abundant in the cytoplasm of terminally differentiated cells, notably, the pyramidal neurons in brain and elongated spermatids in testis. Immunoprecipitation experiments further showed that a kinase activity toward myelin basic protein was associated with Pctaire1 in the adult testis and brain and that its activity was potentially regulated through association with regulatory partner(s). These results suggest that Pctaire1 kinase might have an important role in differentiated cells such as postmitotic neurons and spermatogenic cells.

Animals↗

PubMed1998

The identification and characterization of expression of Pftaire-1, a novel Cdk family member, suggest its function in the mouse testis and nervous system.

We have isolated a murine cDNA encoding for a novel putative Cdk-related protein kinase, which has been named Pftaire-1, by screening a testis cDNA library for new serine/threonine kinases. Pftaire-1 showed 50% and 49% amino acid identity with Cdk5 and Pctaire-3, respectively, and contains the eleven subdomains characteristic of the protein kinases. By northern blot analysis we detected two transcripts of approximately 5.5 and 4.9 kb in size. These transcripts were expressed at low level in all murine tissues tested, except in the brain, testis and embryo, where high expression was detected. Cellular localization of the mRNAs by in situ hybridization analysis shows that Pftaire-1 is expressed in late pachytene spermatocytes in the testis and in post mitotic neuronal cells both in the brain and the embryo, suggesting a role of Pftaire-1 both in the process of meiosis as well as neuron differentiation and/or function.

Amino Acid Sequence↗

PubMed1998

Mouse Odf2 cDNAs consist of evolutionary conserved as well as highly variable sequences and encode outer dense fiber proteins of the sperm tail.

The outer dense fibers (ODF) of the mammalian sperm tail comprise a unique, specialized, and very prominent structure, consisting of nine fibers surrounding the axoneme. The ODF may play an important but as yet undefined role in sperm morphology, integrity and function. Study of the ODF is hampered by insufficient knowledge of their protein composition and the genetic regulation of their synthesis. We report here on the characterization of cDNAs encoding the Odf2 proteins of outer dense fibers of mouse sperm. We isolated two cDNA clones with variable 5' regions. Variability in sequence is restricted to specific regions in the N-terminal part of the encoded proteins, whereas the C-terminal part is highly conserved in Odf2 proteins both between species and within a species. This variability is confirmed at the protein level. The outer dense fibers could be detected immunologically in total sperm tails allowing a direct comparison of their length in relation to the length of the sperm tail. Odf2 transcripts could be demonstrated in testicular RNA and are restricted to germ cells. The start of transcription is in step 5 spermatids of tubular stage V and the RNA could be detected in the cytoplasm of differentiating spermatids in all subsequent tubular stages.

Amino Acid Sequence↗

PubMed1998

Expression and potential role of Fsrg1, a murine bromodomain-containing homologue of the Drosophila gene female sterile homeotic.

We have isolated a cDNA which is a murine homologue of the Drosophila gene female sterile homeotic (fsh). This homologue, which we have designated Fsrg1*, contains two bromodomains and an ET motif characteristic of the Fsh sub-class of bromodomain-containing proteins. Northern blot hybridization analysis of adult tissues revealed that Fsrg1 was expressed at low levels rather ubiquitously, but most abundantly in the testis and ovary. Polyclonal antibodies raised against an Fsrg1 fusion protein were used to characterize the Fsrg1 gene product in tissues. Constructs were also generated in which the Fsrg1 cDNA was tagged with epitopes for hemaglutinin and used in transfection experiments. Immunoblot analysis revealed that the Fsrg1 protein migrates with a relative molecular mass of approximately 110 kDa, although the cDNA sequence would predict a protein of approximately 88 kDa. The migration at approximately 110 kDa was observed for both in vivo protein and protein produced in cultured cells. The Fsrg1 protein was localized to the nucleus when expressed in cultured cells, consistent with the presence of a nuclear localization signal motif in the Fsrg1 sequence. No kinase activity was detected for this nuclear protein as assessed in either autokinase or specific substrate assays. In situ hybridization analysis revealed strikingly high expression of Fsrg1 in granulosa cells of growing follicles in the adult ovary and suggested its possible involvement in folliculogenesis. Additional clues to its potential function were provided by the demonstration of its high level of expression in epithelia of tissues which undergo hormonally-modulated remodeling.

3T3 Cells↗

PubMed1997

Ragweed sensitization alters pulmonary vascular responses to bronchoprovocation in beagle dogs.

In ragweed (RW)-sensitized beagle dogs, we tested the hypothesis that reactivity of the pulmonary vasculature was enhanced with aerosolized histamine (Hist) and RW. Seven dogs were neonatally sensitized with repeated intraperitoneal RW injections, and 12 dogs were controls (Con). The dogs were anesthetized with intravenous chloralose, mechanically ventilated, and instrumented with femoral arterial and pulmonary artery catheters. Specific lung compliance (CLsp), specific lung conductance (Gsp), systemic vascular resistance index, and pulmonary vascular resistance index (PVRI) were measured before and after bronchoprovocation with Hist and RW. After Hist inhalation (5 breaths of 30 mg/ml), both Con and RW dogs had significant (P < 0.05) decreases in CLsp (-51 +/- 4 and -53 +/- 5%, respectively) and Gsp (-65 +/- 5 and -69 +/- 3%, respectively), but only RW-sensitized dogs had a significant increase in PVRI (38 +/- 10%). After RW inhalation (60 breaths of 0.8 mg/ml), only RW-sensitized dogs had significant increases (62 +/- 20%) in PVRI and decreases in Gsp (-77 +/- 4%) and CLsp (-65 +/- 7%). We conclude that, compared with Con, RW-sensitized beagle dogs have increased pulmonary vasoconstrictive responses with Hist or RW inhalation.

Allergens↗

PubMed1997

The NIMA-related kinase 2, Nek2, is expressed in specific stages of the meiotic cell cycle and associates with meiotic chromosomes.

The Aspergillus nimA gene encodes a Ser/Thr protein kinase which is required for mitosis, in addition to Cdc2, and which has been suggested to have a role in chromosomal condensation. In this study, we isolated a potential murine homologue of nimA, Nek2, which was shown to be expressed most abundantly in the testis of the adult tissues examined. Its expression in the testis was restricted to the germ cells, with highest levels detected in spermatocytes at pachytene and diplotene stages. Immunohistochemical analysis revealed that Nek2 localized to nuclei, exhibiting a non-uniform distribution within the nucleus. Nek2 appeared to be associated with meiotic chromosomes, an association that was better defined by immunolocalization to hypotonically dispersed meiotic chromosomes. This localization was more apparent in regions of dense chromatin, including the sex vesicle, and was also obvious at some of the chromosome ends. The presence of Nek2 protein was not unique to male germ cells, as it was found in meiotic pachytene stage oocytes as well. Furthermore, in an in vitro experimental setting in which meiotic chromosome condensation was induced with okadaic acid, a concomitant induction of Nek2 kinase activity was observed. The expression of Nek2 in meiotic prophase is consistent with the hypothesis that in vivo, Nek2 is involved in the G2/M phase transition of the cell cycle. Our results further provide evidence that in vivo, mouse Nek2 is involved in events of meiosis, including but not limited to chromosomal condensation.

Amino Acid Sequence↗

PubMed1996

Compliance and diameter mismatch affect the wall shear rate distribution near an end-to-end anastomosis.

The development of intimal hyperplasia near the anastomosis of a vascular graft to an artery may be related to changes in the wall shear rate distribution. Mismatches in compliance and diameter at the end-to-end anastomosis of a compliant artery and a rigid graft cause shear rate disturbances that may induce intimal hyperplasia and ultimately graft failure. The goal of this study is to determine how compliance mismatch, diameter mismatch, and impedance phase angle affect the wall shear rate distribution in end-to-end anastomosis models under sinusoidal flow conditions. Wall shear rates are obtained through flow visualization using a photochromic dye. In a model with a well-matched graft diameter (6% undersized), the compliance mismatch causes low mean wall shear rates near the distal anastomosis. Considering diameter mismatch, the wall shear rate distributions in 6% undersized, 16% undersized, and 13% oversized graft models are markedly different at similar phase angles. In the two undersized graft models, the minimum mean shear rate occurs near the distal anastomosis, and this minimum is lower in the model with greater diameter mismatch. The oversized graft model has a minimum mean shear rate near the proximal anastomosis. Thus in all three models, the minimum mean wall shear rate is observed at the site of the divergent geometry. The impedance phase angle, which can be altered by disease states and vasoactive drugs, has a minor effect on the wall shear rate amplitude far from the anastomosis but a more pronounced effect closer to the anastomosis. Mean wall shear rates under sinusoidal flow conditions are significantly lower than under steady flow conditions at the same mean flow rate, but they are fairly insensitive to phase angle changes. In order to avoid the divergent geometry that may cause lower wall shear rates, we recommend that compliance mismatch be minimized whenever possible and that graft diameter be chosen to match the arterial diameter at the relevant physiologic pressure, not at the reduced pressure present when the graft is implanted.

Anastomosis, Surgical↗

PubMed1996

Glucocorticoid regulation of a transcription factor that binds an initiator-like element in the murine thymidine kinase (Tk-1) promoter.

Glucocorticoids inhibit transcription of the murine cytoplasmic thymidine kinase gene (Tk-1). Glucocorticoid regulation of Tk-1 transcription can be demonstrated in cells that are arrested in late G1. This observation indicates that inhibition of Tk-1 expression is not dependent upon redistribution within the cell cycle but is due to glucocorticoid regulation of this gene. Transfection studies have been carried out using chimeric genes in which restriction fragments of the Tk-1 promoter were fused to chloramphenicol acetyltransferase or neomycin phosphotransferase. These chimeric reporters were assayed for stable expression and glucocorticoid regulation in P1798 lymphoma cells. A 140-bp fragment, extending from -143 to -3 bp with respect to the thymidine kinase translational start site, was capable of both basal and glucocorticoid-regulated transcription of reporter genes. The extent of inhibition by glucocorticoids was similar to that observed for the endogenous gene, and no increase in basal expression or the extent of inhibition was observed with constructs containing additional 5'-flanking DNA. The 140-bp Tk-1 core promoter fragment binds to transcription factors in extracts from P1798 cells. Control cell extracts contain factors that bind to and protect (from deoxyribonuclease I) a distal promoter element from -106 to -87 bp, relative to the translational start site. A second, proximal element was protected at -43 to -36 bp. The proximal element of the Tk-1 promoter resembles an RNA polymerase II initiator element. No other elements were protected. Glucocorticoids inhibit the amount or activity of the transcription factor that binds to this initiator-like element within the Tk-1 promoter. This element, when fused to upstream activation sequences from the herpes simplex virus thymidine kinase promoter, conveys glucocorticoid sensitivity in cis.

Administration, Topical↗

PubMed1995

c-Myc and cyclin D3 (CcnD3) genes are independent targets for glucocorticoid inhibition of lymphoid cell proliferation.

Glucocorticoids inhibit the expression of critical cell cycle-regulatory genes. The G1 cyclin gene CcnD3, which encodes cyclin D3, is inhibited by dexamethasone in P1798 murine T lymphoma cells. Glucocorticoids also inhibit expression of the catalytic partner of cyclin D3, Cdk4. Inhibition of these two genes results in a decrease in the ability to phosphorylate the Rb-1 tumor suppressor gene product. Stable transformation with SV40 T antigen expression vectors prevents glucocorticoid-mediated cell cycle arrest, which is consistent with the conclusion that glucocorticoids inhibit Rb-1 phosphorylation. Overexpression of cyclin D3 suffices to restore Rb-kinase activity in glucocorticoid-treated cells. Nevertheless, overexpression of cyclin D3 does not prevent glucocorticoid inhibition of cell proliferation. Cells transformed with Cdk4 expression vectors, with or without cyclin D3 expression vectors, also undergo G0 arrest in the presence of dexamethasone. Glucocorticoids inhibit c-Myc expression in lymphoid cells, and transient expression of c-Myc protein attenuates the lytic response in glucocorticoid-treated human leukemia cells (R. Thulasi, D. V. Harbour, and E. B. Thompson, J. Biol. Chem., 268: 18306-16312, 1993). However, P1798 cells stably transfected with c-Myc expression vectors are sensitive to glucocorticoid-mediated G0 arrest. Such transformants withdraw from the cell cycle when treated with dexamethasone. P1798 cells were transformed so as to express both c-Myc protein and cyclin D3 in the presence of glucocorticoids. These Myc/D3 cells continue to proliferate in the presence of dexamethasone, and virtually all of these cells are capable of entering S phase in the presence of the steroid. Rapid apoptotic cell death occurs when wild-type P1798 cells are treated with dexamethasone in serum-free medium. Myc-transformed and cyclin D3-transformed cells also die rapidly when treated with glucocorticoids in the absence of serum. T antigen transformants are resistant to glucocorticoid-mediated apoptosis in serum-free medium. Double transformants that express both cyclin D3 and c-Myc are also resistant to apoptosis in the presence of dexamethasone. We conclude that inhibition of both CcnD3 and c-Myc genes is critical to glucocorticoid-mediated G0 arrest. Furthermore, those genes that convey resistance to growth arrest also convey resistance to cell death.

Animals↗

PubMed1995

Cdk family genes are expressed not only in dividing but also in terminally differentiated mouse germ cells, suggesting their possible function during both cell division and differentiation.

The roles of the cyclin dependent kinase (Cdk) family in murine germ cell development have been examined by studying the expression of five Cdk family genes (Cdc2, Cdk2, Cdk4, Pctaire-1, and Pctaire-3) in mouse reproductive organs. Northern blot and in situ hybridization analyses revealed distinctive expression patterns of these genes with striking cellular, lineage, and developmental stage specificity. We observed Cdk expression in cell types with proliferative activity: Cdc2 and Cdk2 expression in premeiotic spermatocytes in the testis, and Cdc2, Cdk2, and Cdk4 expression in granulosa cells of ovarian follicles. Cdc2 transcripts were most abundant in late pachytene to diplotene spermatocytes, soon to undergo meiosis. Surprisingly, we also observed expression of Cdk family genes in non-proliferating cell types. All five Cdk family genes examined were expressed in Sertoli cells of the adult testis, which are no longer mitotically active. With regard to Pctaire-1 and Pctaire-3, the highest levels of expression were observed in postmeiotic spermatids. Immunoblot analysis also revealed the presence of high levels of Pctaire-1 in postmeiotic germ cells. These results suggest that Cdk family kinases may exhibit various functions in germinal and somatic cells during gametogenesis, not only in the cell cycle but also in other regulatory processes, including differentiation.

Animals↗