Biomedical subjects

J Y Chan

Publications and source records attributed to J Y Chan.

At least 55 records · Page 3Linked to original sources

PubMed1999

Altered expression of the suppressors PML and p53 in glioblastoma cells with the antisense-EGF-receptor.

Gene amplification and enhanced expression of the epidermal growth factor receptor (EGFR) represent the major molecular genetic alteration in glioblastomas and it may play an essential role in cell growth and in the carcinogenic process. On the other hand, the nuclear suppressor proteins PML and p53 are also known to play critical roles in cancer development and in suppressing cell growth. Here we report that, in glioblastoma cells with defective EGFR function, the expressions of both promyelocytic leukaemia (PML) and p53 were altered. Cells that were transfected with the antisense-cDNA of EGFR were found to have more cells in G1 and fewer cells in S phase. In addition, the transfected cells were found to be non-responsive to EGF-induced cell growth. Interestingly, the expression of the suppressors p53 and PML were found to be significantly increased by immunohistochemical assay in the antisense-EGFR cells. Moreover, the PML expression in many of the cells was converted from the nuclear dot pattern into fine-granulated staining pattern. In contrast, the expressions of other cell cycle regulated genes and proto-oncogene, including the cyclin-dependent kinase 4 (cdk4), retinoblastoma, p16INK4a and p21H-ras, were not altered. These data indicate that there are specific inductions of PML and p53 proteins which may account for the increase in G1 and growth arrest in antisense-EGFR treated cells. It also indicates that the EGF, p53 and PML transduction pathways were linked and they may constitute an integral part of an altered growth regulatory programme. The interactions and cross-talks of these critical molecules may be very important in regulating cell growth, differentiation and cellular response to treatment in glioblastomas.

Brain Neoplasms↗

PubMed1999

Hepatitis B envelope protein mutants in human hepatocellular carcinoma tissues.

Hepatitis B virus (HBV) envelope mutants in the region encoding the highly immunogenic major hydrophilic region (MHR) of surface antigen (HBsAg) have been associated with vaccine failure and chronic infection. To determine if these mutants are associated with the development of human hepatocellular carcinoma (HCC), we measured the frequency and nature of such mutants in 23 HBV-associated HCC and various control tissues by performing Southern blot analysis, the polymerase chain reaction (PCR) and direct sequencing. The HBV genome was present mainly in an integrated form and, in most of the samples, the envelope gene was intact. Amino acid substitutions, involving the MHR region in the HCC tissues, were analysed in 11 (61. 1%) of 18 patients with HCC. The mutation Gly145Arg, which has been reported to be associated with immunoevasion, was found in seven of the 18 HCC tissues. A significantly higher frequency of mutations was found in HCC tissues (11 of 18) than in the corresponding non-tumorous tissue of the same patients (one of eight), and in samples from patients with acute (one of 19) or chronic (three of 31) HBV infection (P < 0.001, Fisher's exact test). The accumulation of these envelope mutants in the HCC tissue suggests that such envelope protein mutations may play a role in the process of oncogenesis and that specific vaccines may need to be developed to prevent the occurrence of mutant HBV-associated HCC. Alternatively, the progressive accumulation of mutants in patients with acute hepatitis, chronic hepatitis and HCC may reflect the increased length of duration of HBV infection in these groups of liver lesions.

Adult↗

PubMed1999

Dopaminergic neurotransmission at the paraventricular nucleus of hypothalamus in central regulation of penile erection in the rat.

PURPOSE: To investigate whether the paraventricular nucleus of hypothalamus (PVN) is involved in the central regulation of apomorphine-induced penile erection in the rat, and to decipher dopamine receptor subtypes in the PVN that are involved in apomorphine-induced penile erection. MATERIALS AND METHODS: Male adult Sprague-Dawley rats (200 to 300 gm.) anesthetized with pentobarbital sodium were used. The intracavernous pressure (ICP), recorded along with systemic and mean arterial pressure (SAP, MAP) as well as heart rate (HR), was measured via a 26-gauge needle inserted into one corpus cavernosum. The PVN was activated by stereotaxically delivered apomorphine hydrochloride (0.1 nmol./100 nl.). Injection of saline into PVN served as a vehicle control. To investigate the participation of dopamine receptor subtypes in the PVN on apomorphine-induced penile erection, D1 or D2 receptor antagonist, SCH-23390 (100 pmol./100 nl.) or sulpiride (100 pmol./100 nl.) respectively, was administered into the PVN prior to subcutaneous application of apomorphine (80 microg./kg.). The effects on ICP of microinjection of D1, D2 or D3 receptor agonist, SKF-38393 (200 pmol./100 nl.), lisuride (200 pmol./100 nl.) or 7-hydroxy-DPAT (200 pmol./100 nl.) respectively, into the PVN were also evaluated. RESULTS: The mean resting ICP was 5.2+/-0.4 mm. Hg. Upon administration of apomorphine into the PVN, there was a significant increase in ICP that peaked at 50.7+/-5.3 mm. Hg and persisted for 45.2+/-18.0 minutes after an onset latency of 677.7+/-311.6 seconds. Yawning and teeth gnashing were also observed in most of animals during the period of ICP increase. There was no significant change in SAP, MAP or HR. In addition, there was no elevation in ICP after administration of saline to the PVN or direct injection of apomorphine into the cavernous tissue. Microinjection of D1 or D2 receptor antagonist into the PVN blocked the increase in ICP after subcutaneous administration ofapomorphine. Direct application of D2, but not D1 or D3 receptor agonist into the PVN, on the other hand, increased the ICP. CONCLUSIONS: Our results demonstrate that application of apomorphine to the paraventricular nucleus of hypothalamus elicited penile erection in the rat. Such an increase in ICP to apomorphine was due mainly to activation of the D2 receptor subtype in the PVN. These observations indicate that PVN may be involved in the central regulation of apomorphine-induced penile erection in the rat.

Animals↗

PubMed1999

Glutamatergic projection to RVLM mediates suppression of reflex bradycardia by parabrachial nucleus.

We investigated the role of glutamatergic projection from the parabrachial nucleus (PBN) complex to the rostral ventrolateral medulla (RVLM) in the PBN-induced suppression of reflex bradycardia in adult Sprague-Dawley rats that were maintained under pentobarbital anesthesia. Under stimulus conditions that did not appreciably alter the baseline systemic arterial pressure and heart rate, electrical (10-s train of 0.5-ms pulses, at 10-20 microA and 10-20 Hz) or chemical (L-glutamate, 1 nmol) stimulation of the ventrolateral regions and Köelliker-Fuse (KF) subnucleus of the PBN complex significantly suppressed the reflex bradycardia in response to transient hypertension evoked by phenylephrine (5 micrograms/kg iv). The PBN-induced suppression of reflex bradycardia was appreciably reversed by bilateral microinjection into the RVLM of the N-methyl-D-aspartate (NMDA)-receptor antagonist MK-801 (500 pmol) or the non-NMDA-receptor antagonist 6-cyano-7-nitroquinoxaline-2, 3-dione (50 pmol). Anatomically, most of the retrogradely labeled neurons in the ventrolateral regions and KF subnucleus of the ipsilateral PBN complex after microinjection of fast blue into the RVLM were also immunoreactive to anti-glutamate antiserum. These results suggest that a direct glutamatergic projection to the RVLM from topographically distinct regions of the PBN complex may participate in the suppression of reflex bradycardia via activation of both NMDA and non-NMDA receptors at the RVLM.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1999

Differential roles of NMDA and non-NMDA receptors in synaptic responses of neurons in nucleus tractus solitarii of the rat.

The relative role of N-methyl-D-aspartate (NMDA) and non-NMDA receptors in synaptic responses of neurons in caudal nucleus tractus solitarii (cNTS) was delineated by immunohistochemical and electrophysiologic experiments in rats. Double immunohistochemical staining in in vivo experiments revealed that approximately 80% of cNTS neurons that showed Fos-like immunoreactivity induced by baroreceptor activation were generally also immunoreactive to non-NMDA receptor subunits GluR1 or GluR2. On the other hand, only 20% of Fos-labeled cNTS neurons showed immunoreactivity to NMDA receptor subunits NMDAR1 or NMDAR2. Stimulation of the ipsilateral solitary tract at suprathreshold intensity in slice preparations induced Fos expression in the cNTS and evoked either a single action potential or a complex synaptic response consisting of an initial action potential followed by a secondary slow depolarization. In a majority (70%) of cNTS neurons that exhibited the complex synaptic response, both the initial and secondary components were eliminated reversibly by 6-cyano-7-nitroquinoxaline-2,3-dione (20 microM). This non-NMDA antagonist also inhibited the single action potential manifested by the other population of cNTS neurons. On the other hand, only the secondary slow depolarization was blocked by D(-)-2-amino-5-phosphonopentanoic acid (250 microM) or potentiated by NMDA (1.7 microM). Our results suggested that NMDA and non-NMDA receptors are involved differentially in the synaptic responses of cNTS neurons. Non-NMDA receptors may be distributed predominantly on a majority of the second-order cNTS neurons that may receive primary baroreceptor afferent inputs. On the other hand, NMDA receptors are located primarily on higher-order neurons, which may be connected reciprocally with the second-order cNTS neurons.

2-Amino-5-phosphonovalerate↗

PubMed1998

CD164, a novel sialomucin on CD34(+) and erythroid subsets, is located on human chromosome 6q21.

CD164 is a novel 80- to 90-kD mucin-like molecule expressed by human CD34(+) hematopoietic progenitor cells. Our previous results suggest that this receptor may play a key role in hematopoiesis by facilitating the adhesion of CD34(+) cells to bone marrow stroma and by negatively regulating CD34(+) hematopoietic progenitor cell growth. These functional effects are mediated by at least two spatially distinct epitopes, defined by the monoclonal antibodies (MoAbs), 103B2/9E10 and 105A5. In this report, we show that these MoAbs, together with two other CD164 MoAbs, N6B6 and 67D2, show distinct patterns of reactivity when analyzed on hematopoietic cells from normal human bone marrow, umbilical cord blood, and peripheral blood. Flow cytometric analyses revealed that, on average, 63% to 82% of human bone marrow and 55% to 93% of cord blood CD34(+) cells are CD164(+), with expression of the 105A5 epitope being more variable than that of the other identified epitopes. Extensive multiparameter flow cytometric analyses were performed on cells expressing the 103B2/9E10 functional epitope. These analyses showed that the majority (>90%) of CD34(+) human bone marrow and cord blood cells that were CD38(lo/-) or that coexpressed AC133, CD90(Thy-1), CD117(c-kit), or CD135(FLT-3) were CD164(103B2/9E10)+. This CD164 epitope was generally detected on a significant proportion of CD34(+)CD71(lo/-) or CD34(+)CD33(lo/-) cells. In accord with our previous in vitro progenitor assay data, these phenotypes suggest that the CD164(103B2/9E10) epitope is expressed by a very primitive hematopoietic progenitor cell subset. It is of particular interest to note that the CD34(+)CD164(103B2/9E10)lo/- cells in bone marrow are mainly CD19(+) B-cell precursors, with the CD164(103B2/9E10) epitope subsequently appearing on CD34(lo/-)CD19(+) and CD34(lo/-)CD20(+) B cells in bone marrow, but being virtually absent from B cells in the peripheral blood. Further analyses of the CD34(lo/-)CD164(103B2/9E10)+ subsets indicated that one of the most prominent populations consists of maturing erythroid cells. The expression of the CD164(103B2/9E10) epitope precedes the appearance of the glycophorin C, glycophorin A, and band III erythroid lineage markers but is lost on terminal differentiation of the erythroid cells. Expression of this CD164(103B2/9E10) epitope is also found on developing myelomonocytic cells in bone marrow, being downregulated on mature neutrophils but maintained on monocytes in the peripheral blood. We have extended these studies further by identifying Pl artificial chromosome (PAC) clones containing the CD164 gene and have used these to localize the CD164 gene specifically to human chromosome 6q21.

Animals↗

PubMed1998

Participation of hippocampal formation in negative feedback inhibition of penile erection in the rat.

Detailed information on how the central nervous system regulates penile erection, particularly the inhibitory aspect, is sparse. We observed in Sprague-Dawley rats anesthetized and maintained with chloral hydrate that administration of papaverine (400 microg) directly into the corpora cavernosum of the penis produced an increase in intracavernous pressure (ICP). This elicited experimental index for penile erection was accompanied by a transient increase in the root mean square values, concurrent with a shift in the contribution of Theta (increase) and delta (decrease) power to the hippocampal electroencephalographic (hEEG) activity. Reversal blockade of these hEEG responses with xylocaine, given either intrathecally at the L6-S1 spinal levels or unilaterally to the hippocampal formation, significantly heightened and prolonged the ICP response. Pretreatment with xylocaine by itself, however, did not alter appreciably the baseline ICP or hEEG activity. These results suggest the presence of a novel negative feedback inhibitory mechanism in the hippocampal formation, which is triggered by ascending sensory inputs initiated by tumescence of the penis during normal erectile processes.

Anesthetics, Local↗

PubMed1998

Targeted disruption of the ubiquitous CNC-bZIP transcription factor, Nrf-1, results in anemia and embryonic lethality in mice.

The CNC-basic leucine zipper (CNC-bZIP) family is a subfamily of bZIP proteins identified from independent searches for factors that bind the AP-1-like cis-elements in the beta-globin locus control region. Three members, p45-Nf-e2, Nrf-1 and Nrf-2 have been identified in mammals. Expression of p45-Nf-e2 is largely restricted to hematopoietic cells while Nrf-1 and Nrf-2 are expressed in a wide range of tissues. To determine the function of Nrf-1, targeted disruption of the Nrf-1 gene was carried out. Homozygous Nrf-1 mutant mice are anemic due to a non-cell autonomous defect in definitive erythropoiesis and die in utero.

Amino Acid Sequence↗

PubMed1998

Differential participation of hippocampal formation in cocaine-induced cortical electroencephalographic desynchronization and penile erection in the rat.

We evaluated the role of the hippocampal formation in cocaine-induced cortical electroencephalographic (EEG) desynchronization and penile erection. Adult, male Sprague-Dawley rats anesthetized and maintained by chloral hydrate were used. Intravenous (1.5 or 3.0 mg/kg) administration of cocaine dose-dependently increased intracavernous pressure (ICP), our experimental index for penile erection. This was accompanied by desynchronization of EEG activity recorded from the somatosensory cortex (cEEG), as represented by a decrease in root mean square (RMS) and an increase in mean power frequency (MPF) values. There was a simultaneous increase in the RMS values, without significant changes in the MPF values of EEG signals recorded from the hippocampal formation (hEEG). In animals that received prior application of 10% xylocaine either intrathecally (i.t.) at the L6-S1 spinal levels or directly into the bilateral hippocampal formation, the RMS values of both cEEG and hEEG signals induced by cocaine (1.5 or 3.0 mg/kg, i.v.) were appreciably reduced, along with a further increase in ICP. Unilateral microinjection of cocaine (15 or 30 microg) into the hippocampal formation elicited discernible excitation of both cEEG and hEEG signals. Intriguingly, the ICP underwent a significant and dose-dependent reduction, which was discernibly antagonized by i.t. application of xylocaine. We conclude that cocaine may effect cortical EEG desynchronization but cause a reduction in ICP via an action on the hippocampal formation.

Animals↗

PubMed1998

Mediation by N-methyl-D-aspartate and non-N-methyl-D-aspartate receptors in the expression of Fos protein at the nucleus tractus solitarii in response to baroreceptor activation in the rat.

We investigated the role of glutamatergic synapses in the expression of Fos protein at the nucleus tractus solitarii following baroreceptor activation in rats anaesthetized with pentobarbital sodium. Microinjection of L-glutamate (1 nmol) bilaterally into the nucleus tractus solitarii elicited significant hypotension and bradycardia. There was a concurrent increase, as determined immunohistochemically, in the expression of Fos protein at the commissural, medial and dorsomedial subnuclei of the caudal nucleus tractus solitarii. These effects were blunted when L-glutamate was co-administered with either the selective N-methyl-D-aspartate or non-N-methyl-D-aspartate glutamate receptor antagonist, dizocilpine maleate (200 pmol) or 6-cyano-7-nitroquinoxaline-2,3-dione (8 pmol), into the caudal nucleus tractus solitarii. Repeated and scheduled transient hypertension evoked by phenylephrine (2.5, 5.0 or 10.0 microg/kg, i.v.) also appreciably increased the number of Fos-immunoreactive neurons at the commissural, medial and dorsomedial subnuclei of the caudal nucleus tractus solitarii. The expression of Fos protein in this fashion was reduced, simultaneous with a discernible depression in baroreceptor reflex response, when baroreceptor activation was coupled with microinjection bilaterally of dizocilpine maleate (200 pmol) or 6-cyano-7-nitroquinoxaline-2,3-dione (8 pmol) into the nucleus tractus solitarii. Regression analysis showed that the depressive action on the baroreceptor reflex response by both glutamate receptor antagonists correlated positively to the reduction in Fos-immunoreactivity in the nucleus tractus solitarii after baroreceptor activation. Double immunohistochemical staining revealed that nucleus tractus solitarii neurons that showed Fos immunoreactivity were generally also immunoreactive to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor subunit 1. On the other hand, Fos immunoreactivity was usually absent from neurons in the nucleus tractus solitarii that were immunoreactive to N-methyl-D-aspartate receptor subunit 1. These results suggest that glutamatergic neurotransmission plays an active role, via comparable contributions from both N-methyl-D-aspartate and non-N-methyl-D-aspartate receptors, in the expression of Fos protein at the caudal nucleus tractus solitarii in response to baroreceptor activation.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1998

Altered expression of the growth and transformation suppressor PML gene in human hepatocellular carcinomas and in hepatitis tissues.

The promyelocytic leukaemia (PML) gene, which encodes a transformation and growth suppressor, was first identified at a chromosomal translocation break point in acute promyelocytic leukaemia. To elucidate if PML may be involved in hepatocellular carcinoma (HCC), the expression of PML was analysed using immunohistochemistry in human HCC and hepatitis tissues. Our studies demonstrated overexpression of PML protein in the PML-oncogenic domain (POD) structure in 50% of HCC (11/22). Enhanced expression and cytoplasmic localisation of PML was associated with cirrhosis. Increased expression of PML was also found in liver abscesses. However, in colon metastasis to the liver, the expression of PML was moderate to low, although strong expression was seen in the surrounding interstitial cells, macrophages and lymphocytes, an indication of the inflammation process associated with tumour growth. Most interestingly, strong expression of PML was found in neoplastic cells at the periphery of the tumours, but progressively decreased in cells at the centre of the tumours, which may be associated with an altered transform phenotype or apoptosis. The altered expression of PML indicates that this nuclear protein may play an important role in cellular response to stress and inflammation, as well as in compensatory cell growth.

Carcinoma, Hepatocellular↗

PubMed1998

BRE: a modulator of TNF-alpha action.

A stress-responsive gene highly expressed in brain and reproductive organs (BRE) is down-regulated after UV irradiation, DNA damaging agents, or retinoic acid treatment. The human BRE gene encodes a mRNA of 1.9 kb, which gives rise to a protein of 383 amino acids with a molecular size of 44 kilodaltons. BRE is not homologous to any known gene and its function has not been defined. Here we report that BRE was identified multiple times in a yeast two-hybrid screen of a murine cerebellar cDNA library, using the juxtamembrane domain of the p55 tumor necrosis factor alpha (TNF) receptor. The interaction between the p55 receptor and BRE was verified by an in vitro biochemical assay by using recombinant fusion proteins and by co-immunoprecipitation of transfected mammalian cells. In the yeast two-hybrid assay, BRE specifically interacted with p55 TNF receptor but not with other TNF family members such as the Fas receptor, the p75 TNF receptor, and p75 neurotrophin receptor. Overexpression of BRE inhibited TNF-induced NFkappaB activation, indicating that the interaction of BRE protein with the cytoplasmic region of p55 TNF receptor may modulate signal transduction by TNF-alpha.

Animals↗

PubMed1998

Noradrenergic neurotransmission at PVN in locus ceruleus-induced baroreflex suppression in rats.

We investigated the role of ascending noradrenergic projections from the locus ceruleus (LC) to the paraventricular nucleus (PVN) of the hypothalamus in LC-induced suppression of the baroreceptor reflex (BRR) response in adult Sprague-Dawley rats maintained under pentobarbital anesthesia. On the basis of in vivo microdialysis and high-performance liquid chromatography-electrochemical detection, microinjection of L-glutamate (5 nmol) into the LC resulted in a site-specific increase in norepinephrine (NE) concentration in the dialysate collected from the parvocellular subnucleus of the PVN. The temporal course of this increase in extracellular NE concentration in the PVN coincided with the time course of inhibition elicited by the LC on the BRR response. Microinfusion of NE (10, 50, or 100 nM) into the parvocellular subnucleus of the PVN by reverse microdialysis also promoted a parallel increase in NE at the PVN and a reduction in the BRR response. Inhibition of the BRR response induced by microinjection into the PVN of the alpha 1-adrenoceptor agonist phenylephrine (10 nmol) or chemical activation of the LC was reversed by bilateral PVN microinjection of prazosin (100 pmol). However, local application to the PVN of the alpha 2- or beta-adrenoceptor agonist guanabenz (10 nmol) or isoproterenol (10 nmol) was ineffective. Our results suggest that NE released from the LC-PVN noradrenergic projection may participate in LC-induced suppression of the BRR response by activating the alpha 1-adrenoceptors at the parvocellular subnucleus of the PVN.

Animals↗

PubMed1998

Elevated Fos expression in the nucleus tractus solitarii is associated with reduced baroreflex response in spontaneously hypertensive rats.

We delineated the functional role of Fos protein at the nucleus tractus solitarii in the manifestation of reduced baroreceptor reflex control of heart rate during hypertension, using spontaneously hypertensive rats (SHR), stroke-prone SHR, Wistar-Kyoto rats, or Sprague-Dawley rats. Microinjection into the bilateral nucleus tractus solitarii of an antisense oligonucleotide that targets against the initiation codon of c-fos mRNA significantly potentiated the baroreceptor reflex in response to 30 minutes of sustained increase in blood pressure. Of particular note was the restoration of both the impaired sensitivity and capacity of baroreceptor reflex in SHR and stroke-prone SHR to levels comparable to those in normotensive rats. Likewise, the number of Fos-immunoreactive nuclei evoked by the sustained increase in blood pressure in the caudal nucleus tractus solitarii of SHR and stroke-prone SHR was reduced, after this antisense c-fos treatment, to the basal level exhibited by the normotensive animals. Control treatment with the corresponding sense oligonucleotide, an antisense oligonucleotide that targets against a different portion of the coding sequence of the c-fos mRNA or artificial cerebrospinal fluid, on the other hand, elicited no discernible effect on either the baroreceptor reflex response or the induced expression of Fos protein in the nucleus tractus solitarii by baroreceptor activation. We also found that the basal level of Fos expression in the caudal nucleus tractus solitarii was significantly elevated in the SHR and stroke-prone SHR. Together, these novel findings suggest that an elevated expression of basal Fos protein in the NTS during hypertension may be associated with the dysfunction in baroreceptor reflex control of heart rate.

Animals↗

PubMed1998

In vitro repair synthesis of BCNU-induced DNA damage.

The nitrosoureas including BCNU are potent chemotherapeutic drugs and have been used extensively for treatment of brain tumors and other neoplasias but the mechanisms of action for the DNA lesions created and their repair are still unclear. We have recently determined the in vitro repair of BCNU-treated DNA with cellular extracts and with DNA modifying enzymes. BCNU not only caused an increase in breaks in plasmid DNA, but an increase in cross-linked DNA was also observed after restriction enzyme digestion followed by gel electrophoresis. When HeLa cell-extracts were incubated with BCNU-treated DNA, 5-10 fold increases in DNA repair synthesis were observed as compared with untreated control. Substantial increases in 5'OH and 3'OH sites of the breaks were also found in BCNU-treated DNA as determined by the 10-20 fold increases in labeling with T4-DNA kinase and by endogenous polymerases, while the amount of ligatable sites were at a minimal. When the repair capacity of two glioma cell lines (UWR1 and UWR3) with differential BCNU sensitivity, and cells from a chromosomal breakage disease, Bloom's syndrome (BS), were assessed, the activities of the two glioma cells were about 20-30% of the normal lymphoblastoid cells and HeLa cells, whereas no difference was observed in BS cells. However, differential patterns of DNA bands were observed in the glioma samples suggesting cell-type specific capacities of repair synthesis. These data are in accordance with the concept that BCNU creates multiple DNA lesions and suggests different cell types may develop a variety of repair capabilities.

Bloom Syndrome↗

PubMed1997

Cell-cycle regulation of DNA damage-induced expression of the suppressor gene PML.

The promyelocytic leukemia (PML) gene, which encodes a growth- and transformation-suppressor, has been identified at the non-random chromosomal translocation break point t(15; 17)(q22; q12) of acute promyelocytic leukemia. To elucidate if PML may play a role in cellular response to DNA damage, PML expression was analyzed by immunofluorescence staining in HeLa cells treated with ionizing radiation (IR) and cisplatin. Our studies demonstrated IR at 20Gy, and cisplatin at 6 micrograms/ml caused more than 5-10 fold increases in PML protein expression in the PML Oncogenic Domain (POD) by immunofluorescent staining. Northern blotting showed that there was no gross increase in mRNA levels indicating that the induction is a post-transcriptional event. Flow cytometry showed that HeLa cells treated with IR were progressively arrested in G1, which correlates with the optimal expression of PML in the cell cycle. To determine if PML expression was under the control of the tumor suppressor p53, which is known to arrest cells in G1, HeLa cells were transfected with the wild-type p53 gene. PML expression in p53 transduced cells were 5-10 fold higher than the control, indicating that the enhanced expression of PML is apparently dependent on the p53 pathway. These data also indicate that PML may play an important role in cellular response to DNA damage such as DNA repair or apoptosis during G1 arrest.

Blotting, Northern↗

PubMed1997

Participation of paraventricular nucleus of hypothalamus in central regulation of penile erection in the rat.

PURPOSE: To investigate the possible participation of the paraventricular nucleus of hypothalamus in central regulation of penile erection. MATERIALS AND METHODS: Male adult Sprague-Dawley rats were anesthetized and maintained with pentobarbital sodium. The intracavernous pressure (ICP) was used as an experimental index for penile erection, and was recorded alongside systemic arterial pressure and heart rate. The effect on ICP of electrical (30-s train of 30-120 microA, 40-160 Hz, 1-ms rectangular pulses) or chemical (L-glutamate, 0.5 nmol/50 nl.) activation of the paraventricular nucleus of hypothalamus (PVN) or hippocampal formation was evaluated. RESULTS: Electrical activation of the PVN elicited both multiple and single episodes of elevation in ICP, along with visible erection and ejaculation. The former pattern exhibited an increase in ICP that was more sustained, with higher peak amplitude and longer latency. Chemical stimulation of neuronal perikarya in the PVN also resulted in similar patterns of rise in ICP and visible erection. These effects were, nonetheless, not accompanied by significant alterations in systemic arterial pressure and heart rate. Activation of the hippocampal formation, as we reported previously, similarly elicited multiple and single episodes of increase in ICP. These erectile responses, however, were substantially reduced or eliminated upon electrolytic lesion of the ipsilateral PVN. CONCLUSION: These observations suggest that the PVN may be an important nucleus that participates in central neural regulation of penile erection in the rat. Furthermore, an efferent pathway(s) from the hippocampal formation to PVN may constitute part of the neural circuitry in the forebrain in the regulation of erectile functions.

Animals↗

PubMed1997

Involvement of AT2 receptors at NRVL in tonic baroreflex suppression by endogenous angiotensins.

We evaluated the role of endogenous angiotensin II and III (ANG II and ANG III) at the rostral nucleus reticularis ventrolateralis (NRVL) in the modulation of baroreceptor reflex (BRR) response and the subtype of angiotensin receptors involved in this process. Adult male Sprague-Dawley rats anesthetized and maintained with pentobarbital sodium were used. Exogenous application of ANG II or ANG III (10, 20, or 40 pmol) by bilateral microinjection into the NRVL significantly suppressed the BRR response to transient hypertension induced by phenylephrine (5 micrograms/kg i.v.). The suppressive effect of ANG II (20 pmol) was reversed by an equimolar dose (1.6 nmol) of its peptide antagonist, [Sar1, Ile8]ANG II, and the nonpeptide antagonists for AT1 and AT2 receptors, losartan and PD-123319, respectively. On the other hand, the inhibitory action of ANG III (20 pmol) was blunted by its peptide antagonist. [Ile7]ANG III or PD-123319, but not by losartan. Blocking the endogenous activity of the angiotensins by microinjection into the bilateral NRVL of [Sar1, Ile8]ANG II, [Ile7]ANG III, or PD-123319 elicited an appreciable enhancement of the BRR response, whereas losartan produced minimal effect. These results suggest that, under physiological conditions, both endogenous ANG II and ANG III may exert a tonic inhibitory modulation on the BRR response by acting selectively on the AT2 receptors at the NRVL.

Angiotensin II↗