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Biomedical subjects

C Y Cheung

Publications and source records attributed to C Y Cheung.

At least 91 records · Page 5Linked to original sources
PubMed1984

Does beta-endorphin modulate basal and dopamine-inhibited prolactin release by an action at the anterior pituitary?

Previous studies performed to elucidate an anterior pituitary site of action for beta-endorphin (B-EP) in the regulation of prolactin (PRL) release have been inconclusive. The purpose of the present study was to further investigate if B-EP could stimulate PRL release by a direct action at the anterior pituitary. Anterior pituitaries from ovariectomized rats were dispersed, and the cells were plated in Ham's F-10 medium. Following a 48-hour preincubation period, the effects of B-EP on basal PRL release and on the dopamine-inhibited PRL release were studied over a 24-hour period. Overall, B-EP at concentrations ranging from 10(-8) to 10(-6) M had no effect on basal release of PRL at 1, 3, 5 and 24 h, although a small but significant increase of 39% was observed with 10(-6) M B-EP at 1 h of incubation only. B-EP at 10(-6) M, but not at lower concentrations, significantly suppressed the inhibitory effects of 10(-6) M dopamine on PRL release at 1 h but not at longer periods of incubation. Pretreatment of the cells for 3 h with 10(-6) M naloxone antagonized the suppressive effect of B-EP on the dopamine-mediated inhibition of PRL release. Naloxone pretreatment of the cells had no effect on basal nor dopamine-inhibited release of PRL. Because of the high concentration of B-EP required to suppress the dopamine inhibition and the transitory nature of this B-EP effect, it is unlikely that the primary site of action for B-EP in regulating PRL release is at the anterior pituitary.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Dopaminergic regulation of prolactin and growth hormone synthesis in rat pituitary tumor cells.

The dopaminergic regulation of prolactin (PRL) and growth hormone (GH) synthesis were studied in monolayer cultures of rat pituitary tumor (GH3) cells. PRL synthesis could not be inhibited by dopamine at concentrations ranging from 10(-9) to 10(-5) M. Bromocriptine inhibited PRL synthesis only at the very high concentration of 10(-5) M, and this effect was not reversible by metoclopramide. GH synthesis was inhibited by 10(-6) and 10(-5) M dopamine but not by bromocriptine, and the dopamine effect was blocked by metoclopramide. These results suggest the existence of a defective dopaminergic regulation of PRL and GH synthesis in GH3 cells.

Animals↗
PubMed1983

Prolactin suppresses luteinizing hormone secretion and pituitary responsiveness to luteinizing hormone-releasing hormone by a direct action at the anterior pituitary.

In pathological or experimental hyperprolactinemia, the elevated circulating levels of PRL are the usual cause of the impairment in gonadotropic function. The present study was undertaken to determine whether PRL could suppress basal LH secretion and LHRH-stimulated LH release by a direct action at the anterior pituitary. Anterior pituitaries from ovariectomized rats were incubated in medium 199 alone or in medium 199 containing ovine PRL, and basal and the LHRH-stimulated LH release were followed for 2 or 3 h in vitro. Ovine PRL at 40 and 80 micrograms/ml suppressed basal LH release by 41% and 72%, respectively, at 2 h of incubation. This suppressive effect of both concentrations of PRL continued to the third hour of incubation. LHRH at 5 ng/ml increased the release of LH from pituitaries incubated in medium alone by 57%, 61%, and 107% at 1, 2, and 3 h of incubation, respectively. However, in the pituitaries treated with 40 micrograms/ml ovine PRL, the stimulatory effects of LHRH were diminished at all time points measured. Pretreatment of anterior pituitaries with ovine PRL for 6 h significantly inhibited by 81% the LHRH (5 ng/ml) stimulation of LH release at 2 h of incubation. On the other hand, inhibition of endogenous PRL release by 10(-6) M bromocriptine enhanced the stimulatory effects of 5 ng/ml LHRH by 2.5-fold at 2 h of incubation. The inhibitory effects of PRL on basal and stimulated LH secretion appeared unique, since neither BSA nor vasopressin could elicit similar suppressive effects on LH. These results suggest that in anterior pituitaries exposed to elevated levels of PRL, LH secretion and pituitary responsiveness to LHRH could be impaired. This phenomenon may contribute in part to the antigonadotropic effects of PRL.

Animals↗
PubMed1982

Mammotroph and gonadotroph volume percentage in the rat anterior pituitary after lesions of the medial basal hypothalamus.

A lesion of the medial basal hypothalamus (MBH) results in enhanced secretion of prolactin and reduced release of the other hormones from the anterior pituitary. We measured the volume percentage of the prolactin-secreting mammotrophs, stained immunocytochemically, under several experimental conditions to assess the morphological correlated of the high secretion of prolactin. In addition, gonadotrophs were stained both tinctorially with the periodic acid-Schiff reagent and immunocytochemically with antibodies to luteinizing hormone. Anterior pituitaries were studied from female rats which were: (1) cycling, (2) ovariectomized for 2 weeks, (3) ovariectomized for 2 weeks with a lesion of the MBH for 1, 14 and 21 days. The volume percentage of mammotrophs and gonadotrophs in the lateral, central and middle fields of the anterior pituitary was determined and this measurement was expressed relative to endocrine cellular area so that vascular space and necrotic regions would not be included in the assessment. Ovariectomy significantly decreased mammotroph and increased gonadotroph volume percentage when compared to the intact animals. MBH lesions had the opposite effect. At 14 and 21 days after a lesion, the volume percentage of mammotrophs was significantly greater than in the intact rat, while the volume percentage of gonadotrophs was identical to that observed in the intact rat. These data suggest that the elimination of the prolactin-inhibitory hormone(s) of the MBH result in hypertrophy and/or hyperplasia of mammotrophs. This effect occurs in the absence of ovarian and may be of importance in the induction of prolactin-secreting pituitary adenomas. The data also support the hypothesis that destruction of gonadotropin-releasing hormone neuron terminals countermands the hypertrophy of gonadotrophs induced by ovariectomy.

Animals↗
PubMed1981

Stimulation of prolactin secretion by neuroleptics in rats with medial basal hypothalamic lesions.

The ability of dopamine receptor antagonists to stimulate prolactin release in rats with medial basal hypothalamic lesions was investigated. Starting levels of prolactin were elevated to approximately 400 ng/ml in animals in which the tuberoinfundibular dopaminergic neurons were completely destroyed by the lesion. In lesioned animals, chlorpromazine administration at doses of 0.1 and 5 mg/kg induced a further 2- to 3-fold significant increase in plasma prolactin levels. the incubation of anterior pituitaries from lesioned animals with 10(6) M chlorpromazine had no effect on prolactin secretion, thereby eliminating the possibility that chlorpromazine itself stimulates prolactin release from the anterior pituitary. A similar increase in plasma prolactin in lesioned rats was also observed with the potent dopamine antagonist d-butaclamol (1 mg/kg). The effect was stereospecific since the inactive isomer l-butaclamol did not produce any change in the circulating levels of prolactin. Pimozide, another dopaminergic antagonist, was ineffective in inducing a further increase in prolactin in lesioned rats when 0.63 mg/kg was used. However, at a 10-fold lower concentration (0.63 mg/kg), pimozide stimulated a significant increase in prolactin in lesioned rats. The involvement of an alpha-adrenergic mechanism was ruled out by the inability of phentolamine (2.5 mg/kg) to increase prolactin secretion. These data suggest that dopaminergic antagonists can further increase prolactin levels in medial basal hypothalamus-lesioned rats possibly by blocking the inhibitory action of dopamine from nonhypothalamic sources, or by releasing a substance which possesses prolactin-releasing activity.

Animals↗
PubMed1981

Increased responsiveness of the dopamine-mediated inhibition of prolactin synthesis after destruction of the medial basal hypothalamus.

Long term destruction of the tuberoinfundibular dopaminergic neurons results in increased responsiveness of the anterior pituitary to the suppression of PRL release by dopamine. In the present study we tested whether long term destruction of these neurons by medial basal hypothalamic lesions would also increase the potency of dopamine in suppressing the release and apparent synthesis of PRL. Anterior pituitaries from ovariectomized control rats and ovariectomized rats lesioned for 2 weeks were incubated for 3 h in medium 199 containing [3H]leucine in the presence or absence of various concentrations of dopamine. Labeled PRL was separated by polyacrylamide gel electrophoresis and quantitated by liquid scintillation spectroscopy. Concentrations of dopamine ranging from 10(-8)-10(-6) M caused a dose-dependent suppression of labeled newly synthesized PRL released into the medium from anterior pituitaries of medial basal hypothalamus-lesioned animals. With pituitaries from the nonlesioned rats, newly synthesized PRL release was progressively inhibited by 10(-7) and 10(-6) M dopamine, while 10(-8) M dopamine actually significantly stimulated PRL release. Total labeled PRL (that released and that remaining in the gland) was equally suppressed by all three concentrations of dopamine in pituitaries from lesioned animals, but only a minor effect was observed at the highest concentration of dopamine with the control pituitaries. Therefore, the potency of dopamine to suppress the release and apparent synthesis (total labeled PRL) of newly synthesized PRL from pituitaries of long term lesioned animals was increased.

Animals↗
PubMed1977

Effects of testosterone implants and hypothalamic lesions on luteinizing hormone regulation in the castrated male rat.

The effects of intrahypothalamic and subcutaneous implants of testosterone (T) and those of hypothalamic lesions on resting levels of circulating LH and pituitary responsiveness to exogenous LHRH were studied in castrated male rats to elucidate hypothalamic and pituitary regulation of LH secretion. Two hundred mug implants of testosterone propionate (TP) in the median eminence region suppressed plasma LH titers before evidence of direct inhibition of pituitary function (as indicated by testing with LHRH) was found. Such implants release appreciable amounts of T into the peripheral circulation in the immediate post-operative period, and SC Silastic (constant release) capsules containing T have similar effects. The findings suggest that, regardless of the site of implant, the initial negative feedback inhibition of LH by T is not dependent on direct action at the pituitary levels but rather appears to be a hypothalamic effect. In the days following exposure to hypothalamic or peripheral implantation of T, however, a progressively developing decline in the response to exogenous LHRH was observed. In order to determine whether this effect results from suppression of endogenous LHRH release, the median eminence-arcuate region was destroyed to remove the source of LHRH. In these animals, the suppression of plasma LH was evident on the first day after the lesion, but pituitary responsiveness to LHRH was unaffected until after one week. When Sialastic capsules were implanted SC into lesioned animals, a more rapid (less than 1 week) inhibition of pituitary responsivity ensued. Suprachiasmatic lesions did not affect basal LH secretion or pituitary responses to LHRH. The data provide evidence for a dual feedback action of T on LH in castrated male rats: an initial inhibitory effect presumably due to hypothalamic inhibition (commencing at around 6h after hypothalamic of SC implantation of T), and a subsequent suppression of pituitary responisveness (after one day) presumably due to direct action of T on the pituitary. In addition to these phenomena, findings in rats bearing median eminence-arcurate lesions suggest that the removal of endogenous LHRH by itself leads to an eventual decline in pituitary responsiveness (greater than one week postoperatively).

Animals↗
PubMed1976

Supersensitivity of anterior pituitary dopamine receptors involved in the inhibition of prolactin secretion following destruction of the medial basal hypothalamus.

The medial basal hypothalamus of ovariectomized rats was destroyed using a modified Halász knife. Large increases in prolactin secretion were observed 1 and 14 days following the lesions. Long- and short-term lesioned animals were anesthetized with chloral hydrate and treated with various doses of apomorphine (0.05, 0.2, 2, 5 mg/kg). Blood samples were obtained before and 10, 30 and 60 minutes after the injection. Both the 0.05 and 0.2 mg/kg doses caused significantly greater and longer-lasting inhibition of prolactin in long-term than in short-term lesioned animals. Since the MBH was totally destroyed this study suggests that anterior pituitary dopamine receptors involved in the inhibition of prolactin secretion become supersensitive in long-term lesioned rats.

Animals↗
PubMedDate not supplied

Fetal adrenal VIP: distribution and effect on medullary catecholamine secretion.

Vasoactive intestinal peptide (VIP) was found in the adrenal gland of ovine fetuses at 130-135 days gestation and was shown to stimulate catecholamine secretion. VIP was demonstrated by immunocytochemistry using the indirect antibody-enzyme method. VIP-immunoreactive nerve fibers were observed in the capsule, zona glomerulosa and inner layer of the cortex as well as in the medulla; furthermore small clusters of VIP-containing cell bodies were found at the corticomedullary border. To study the direct effect of VIP on catecholamine release, fetal adrenal medulla was dispersed into single cells and incubated in vitro with VIP for 6 hours. Catecholamine release into the medium was measured at 1, 3 and 6 hours. At 6 hours of incubation, VIP stimulated total catecholamine release from fetal adrenomedullary cells in a dose-dependent manner at concentrations ranging from 10(-8) to 10(-4) M. The release of norepinephrine and epinephrine, but not dopamine, was significantly enhanced. The presence of VIP in the fetal adrenal cortex and medulla, and the ability of VIP to stimulate catecholamine release from fetal adrenomedullary cells in vitro suggest that VIP may be an important modulator of medullary catecholamine secretion during fetal life.

Adrenal Glands↗
PubMedDate not supplied

Ontogeny of adrenal VIP content and release from adrenocortical cells in the ovine fetus.

The present study was designed to investigate the presence of VIP in fetal adrenals, to determine the changes in adrenal VIP content associated with maturation, and to explore the factors which regulate fetal adrenal VIP release. Adrenal glands from ovine fetuses at 70 to 140 days gestation were used. Adrenal VIP content, as measured by radioimmunoassay, were low at 70 and 80 days of gestation. This was followed by a rapid increase in VIP content from 80 to 110 days reaching a plateau between 110 and 130 days at levels comparable to that in the adult. A significant fall in adrenal VIP content occurred at 140 days, immediately prior to term. Release of VIP from fetal adrenocortical cells in vitro was significantly elevated by angiotensin II at 10(-5) M, while ACTH had no effect. Acetylcholine at 50 microM and high potassium stimulated fetal adrenal VIP release while norepinephrine did not. These results suggest that the VIP neuronal system in the ovine fetal adrenal matures between 80 and 110 days of gestation. Furthermore, the release of VIP from the fetal adrenocortical cells may be regulated by angiotensin II and cholinergic neurotransmitters.

Acetylcholine↗
PubMedDate not supplied

Developmental expression of vascular endothelial growth factor and its receptors in ovine placenta and fetal membranes.

OBJECTIVES: Vascularity of the surface of the placenta in humans and of the placenta and fetal membranes in several species including sheep is an important determinant of intramembranous absorption of amniotic fluid. Our previous studies have shown that the total blood vessel surface area in ovine amnion and chorion increases with advancing gestation. Vascular endothelial growth factor (VEGF) is a potent angiogenic and permeability factor and is found to be expressed in the ovine placenta and fetal membranes. To investigate the role of VEGF in maintaining the absorptive function of the intramembranous microvessels, the present study was undertaken to determine the gestational change in gene expression of VEGF and its receptors, kinase insert domain-containing receptor (KDR) and fms-like tyrosine kinase (Flt-1), in ovine placenta, chorion, and amnion. METHODS: Total RNA was extracted from placental cotyledon, chorion, and amnion of ovine fetuses at 60-140 days of gestation. The relative abundance of VEGF, KDR, and Flt-1 mRNA was determined by Northern blot analysis, and VEGF molecular forms expressed were identified by reverse transcriptase polymerase chain reaction. The gestational changes in mRNA levels of VEGF and its receptors were analyzed by regression analysis. RESULTS: In ovine placenta, chorion, and amnion, VEGF mRNA levels increased significantly from 60 to 140 days. The major VEGF molecular form expressed in these tissues was VEGF164, whereas VEGF120, VEGF144, and VEGF188 were present at lower levels. In the placenta, KDR was the primary VEGF receptor expressed, although Flt-1 was also detected at very low levels. In the amnion and chorion, KDR was the only receptor expressed. A gestational-dependent change in VEGF receptor expression was not observed in the placenta and membranes. CONCLUSIONS: The increase in VEGF gene expression with advancing gestation in the amnion and chorion where KDR is expressed suggests that VEGF and its receptor are important determinants of vascularity and permeability, and thus exchange capacity, of the intramembranous pathway.

Amnion↗
PubMedDate not supplied

A pH-sensitive polymer that enhances cationic lipid-mediated gene transfer.

The efficient release of nonviral gene carriers from endosomes is an important step for the successful delivery of DNA into the cell nucleus. A synthetic pH-sensitive anionic polymer, poly(propylacrylic acid) (PPAA), was designed to aid in endosomal escape of nonviral vectors and improve the transfection efficiencies with these vectors. Transfection of NIH3T3 fibroblasts with ternary physical mixtures of the cationic lipid DOTAP, pCMVbeta plasmid DNA, and PPAA showed marked enhancement of both gene expression levels and fraction of cells transfected compared to binary control mixtures of DOTAP and DNA. PPAA also significantly improved the serum-stability of DOTAP/DNA vectors. The DOTAP/DNA/PPAA vectors maintained high levels of transfection in media containing up to 50% serum. The striking enhancement of transfection efficiency with cationic lipid/DNA/PPAA mixtures, along with the enhanced serum-stability, suggests that PPAA may provide significant improvements for the in vivo intracellular delivery of drugs such as DNA, oligonucleotides, proteins, and peptides.

3T3 Cells↗
PubMedDate not supplied

Cellular localization of vascular endothelial growth factor in ovine placenta and fetal membranes.

To further understand the role of vascular endothelial growth factor (VEGF) in mediating angiogenesis and vascular permeability during development in the sheep placenta and fetal membranes, we examined the localization of VEGF mRNA and protein in placental, chorionic and amniotic tissues by in situ hybridization and immunohistochemistry in ovine fetuses at 62, 102 and 141 days gestation (term=150 days). In the placenta, VEGF mRNA expression and VEGF protein immunostaining were strong in cytotrophoblasts surrounding the villi. In addition, VEGF protein was localized in smooth muscle cells around fetal and maternal blood vessels and in the maternal epithelium. There was no apparent difference in placental VEGF mRNA or protein levels associated with advancing gestation. In the fetal membranes, VEGF mRNA was detected in the amniotic epithelium and the chorionic cytotrophoblastic cell layer. The intensity of the hybridization signals in both amnion and chorion appeared low at 62 days, moderate at 102 days and high at 141 days gestation. VEGF protein was detected in amniotic epithelium and chorionic cytotrophoblasts at all gestational ages studied. The increase in VEGF gene expression in fetal membranes as term approaches suggests that during fetal development VEGF may promote the vascularity and permeability of the microvessels which perfuse the fetal membranes, as well as permeability of the amniotic membrane itself. Thus VEGF may participate in the regulation of amniotic fluid volume.

Amnion↗
PubMedDate not supplied

Stimulation of HCO3- secretion across cystic fibrosis pancreatic duct cells by extracellular ATP.

Previous studies have demonstrated that extracellular ATP is able to activate Ca2+-dependent Cl- channels which may be important in circumventing the defective cAMP-dependent pancreatic ductal HCO3- secretion in cystic fibrosis (CF). The present study further investigated the effect of extracellular ATP on the stimulation of HCO3- secretion across CF pancreatic duct cells (CFPAC-1). Cells were grown in culture plate inserts which enabled the access of a pH microelectrode to the apical compartment. Changes in apical pH upon stimulation by extracellular ATP, as an indication of HCO3- secretion, were measured. ATP induced a rapid increase in apical pH, which reached a plateau with an averaged pH unit of 0.3 higher than that measured in unstimulated cells. The effect of ATP was concentration dependent. The ATP-induced change in pH could be blocked by apical addition of Cl- channel blockers, indicating that activation of apical Cl- channel is vital for HCO3- secretion by the pancreatic duct cells. Together with the previous finding, the present study suggests that HCO3- secretion may be stimulated in CF pancreatic duct cells by extracellular ATP via a cAMP-independent pathway.

Adenosine Triphosphate↗