Biomedical subjects

J T Clark

Publications and source records attributed to J T Clark.

At least 37 records · Page 2Linked to original sources

PubMed1989

Polymorphism of a 35-48 kDa Plasmodium falciparum merozoite surface antigen.

Glycoproteins of the asexual blood stages of Plasmodium falciparum were labelled with radioactive glucosamine and analysed by two-dimensional electrophoresis. Four major glycoproteins were detected in all eight parasite isolates studied. Two of the glycoproteins, designated GP2 and GP4, were invariant among the isolates, while the other two GP1 and GP3 were found to be polymorphic in both their biochemical and antigenic properties. By immunoblotting and immunoprecipitation with specific monoclonal antibodies, the two polymorphic glycoproteins were identified as surface antigens of merozoites.

Animals↗

PubMed1988

Chronic morphine and testosterone treatment: effects on norepinephrine and serotonin metabolism and gonadotropin secretion in male rats.

The effects of sustained-release implants of morphine (M) and/or testosterone (T) on serum gonadotropin levels and norepinephrine (NE) and serotonin (5-HT) metabolism in brain regions were examined. While 4 days of M or 5 mm [corrected] T treatment were without significant effect, the combination dramatically decreased circulating levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Castration increased NE content of the mediobasal hypothalamus (MBH), and although M, 5 mm [corrected] T, or their combination significantly reduced NE levels in the MBH, they remained elevated compared to intact or 30 mm [corrected] T-treated rats. Further, in the MBH no differences in normetanephrine (NME) levels, nor in NME:NE ratios, nor 5-HT metabolism were evident. In the preoptic area-anterior hypothalamus, NE or 5-HT metabolism were not altered by castration and M and/or T. These results show that M and T interact to suppress LH and FSH release in the apparent absence of any appreciable effect on hypothalamic monoamines.

Animals↗

PubMed1988

Control of feeding and sexual behaviors by neuropeptide Y: physiological implications.

Recent evidence suggests that a variety of hypothalamic neuropeptides may mediate interneuronal communication to coordinate diverse neuroendocrine and behavioral functions. In this work, we describe the effects of neuropeptide Y (NPY) on feeding and sexual behaviors. We observed that central administration of bolus NPY stimulated a robust, dose-related feeding response in satiated male and female rats. Continuous NPY receptor activation also evoked dose-related, intermittent feeding in a manner normally observed during nocturnal feeding. It appears that the paraventricular nucleus in the hypothalamus may be the primary site of NPY action because the anticipated reciprocal changes in NPY concentrations, in response to food deprivation followed by ad libitum food intake, occurred only in this site. Additional findings revealed that NPY-induced feeding may follow either substantial reduction or complete restraint of an inhibitory influence on feeding mediated by alpha 2-adrenoreceptor systems in satiated rats. Further, NPY was found to suppress male and female sexual behaviors. The suppressive effects on sexual behavior were apparent prior to or at the time of the onset of feeding after NPY administration. These observations may provide a neurochemical basis for clinical and animal studies on disorders of feeding associated with diminished reproductive functions.

Animals↗

PubMed1988

Triton-labile antigens in flagella isolated from Giardia lamblia.

Sheared flagella from Giardia lamblia were freed from cytoskeleton fragments and other cell contaminants by centrifuging in a density gradient. The purified organelles contain many polypeptides, including a set of low-molecular-weight antigens [apparent molecular weights (MWs) = 31, 32, 34, 35 and 37 kD] in the same size range as the approximately 30 kD structural giardins of the cytoskeleton. However, on sodium dodecyl sulphate-polyacrylamide gels, the mobilities of individual flagellar polypeptides do not correspond exactly to the cytoskeleton bands, and, unlike the cytoskeleton proteins, the flagellar components are easily extracted by Triton demembranation. The pattern of flagellar isoforms after isoelectric focussing (IEF) and electrophoresis in two dimensions is also clearly different from that of the cytoskeleton proteins. The fact that at least some approximately 30 kD flagellar antigens are localised by immunofluorescence specifically in the two ventral flagella suggests that these proteins may be components of the paraflagellar structures found beneath the membrane of these organelles. In electron micrographs of the isolated flagella, the paraflagellar rods are seen to bridge the membrane to three adjacent doublet microtubules of the axoneme.

Animals↗

PubMed1988

Alpha 2-adrenoceptor blockade attenuates feeding behavior induced by neuropeptide Y and epinephrine.

Neuropeptide Y (NPY, 0.47 nmol) and epinephrine (28.9 nmol) evoked robust, and quantitatively similar, increments in food intake and local eating rate following administration into the third cerebral ventricle (IIIV). Whereas IIIV pretreatment with phentolamine (71 nmol), a nonselective alpha-adrenoceptor antagonist, or prazosin (9.5 nmol), a selective alpha 1-adrenoceptor antagonist, was without effect on NPY-induced feeding behavior, pretreatment with the alpha 2-adrenoceptor antagonist yohimbine (15 nmol) dramatically attenuated the stimulatory effects of NPY or epinephrine on both food intake (by over 50%) and local eating rate. Additionally, yohimbine administered alone was associated with a stimulatory effect on food intake for the periods of 80-110, and 110-140 minutes posttreatment. These data demonstrate that feeding behavior induced by IIIV administration of NPY or epinephrine is attenuated by prior blockade of alpha 2-adrenoceptors and suggest that, as in other systems innervated by neurons displaying NPY and adrenergic transmitter colocalization, the effects of NPY on feeding behavior may, at least in part, be mediated via alpha 2-adrenoceptors.

Adrenergic alpha-Antagonists↗

PubMed1988

Relation of autogrooming to sexual behavior in male rats.

Grooming and penile reflexes were studied in male rats that were restrained in supine position with the penile sheath retracted or were free to copulate with sexually receptive females. In Experiment 1 there was a reliable concordance in supine males between the tendency to groom and the tendency to display penile reflexes. In Experiment 2 we analyzed the sequential organization of grooming and genital events in supine tests. It was assumed that many or most episodes of ventral grooming would have been genital grooming had access to the genitalia not been prevented by restraint. Paw grooming tended to precede clusters of penile responses, whereas ventral grooming started after the onset of erections. Experiment 3 was an exploration of grooming in the context of copulation, rather than supine restraint. Males groomed their genitalia immediately after all intromissions and after all mounts that ended mount bouts. The duration of grooming was not affected by whether or not intromission occurred. Finally, in Experiment 4 we observed genital and nongenital grooming and recorded electromyographic (EMG) activity from the striated bulbospongiosus muscle (mBS) of the penis in freely moving rats. Consistently, mBS activity led to genital grooming with a short latency, whereas nongenital grooming rarely led to genital grooming, and EMG activity was not associated with nongenital grooming nor did it tend to follow after genital grooming was initiated.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Chronic morphine and testosterone treatment. Effects on sexual behavior and dopamine metabolism in male rats.

The effects of sustained delivery of morphine and/or testosterone (T) on male rat copulatory behavior, penile reflexes and dopaminergic metabolism in selected brain regions were examined. Castration was followed by (1) a decrease in the number of male rats exhibiting intromissive and ejaculatory behavior in mating tests, (2) decreased erections in ex copula tests, and (3) increases in dopamine and dihydroxyphenylacetic acid (DOPAC) concentrations in the mediobasal hypothalamus (MBH) and the preoptic area-anterior hypothalamus (POA-AH). The decreased incidence of copulatory behavior and penile reflexes seen after castration was effectively prevented by a 4-day treatment with 5-mm T-containing Silastic capsules. Chronic morphine implants, conversely, accentuated the castration-induced decrements in copulatory behavior and prevented the 5-mm-T-induced facilitation, but did not alter the number of animals displaying erection (although the number of erections displayed by testosterone-treated rats was reduced) in ex copula tests. Treatment of castrated rats with 5 mm T, but not morphine alone, nor the combination of 5 mm T plus morphine, significantly reduced dopamine and DOPAC levels in the MBH. In the POA-AH, 5 mm T was without effect, whereas morphine, alone or in combination with 5 mm T, reduced the levels of dopamine and DOPAC. These data suggest that (1) the decline in sexual behavior induced by chronic morphine is primarily due to a failure of sexual arousal, and not of erectile ability, and (2) although the decline in sexual activity seen after castration is associated with alterations in dopaminergic metabolism, the effects of morphine and testosterone on sexual activity are opposite and dissociated from alterations in dopaminergic metabolism.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1987

Identification and characterisation of proteins associated with the rhoptry organelles of Plasmodium falciparum merozoites.

We describe antigens of Plasmodium falciparum recognised by murine monoclonal antibodies which by immunofluorescence react with the rhoptry organelles of the extracellular merozoite stage. Immunoblotting shows that the antibodies recognise two major parasite antigens of Mr 82 and 65 kilodaltons (kDa). Immunoprecipitations from detergent extracts of [35S]-methionine-labelled parasites show that the 82-kDa and 65-kDa antigens are parasite proteins. Pulse-chase experiments on synchronous parasite cultures show that the 82-kDa protein is synthesised during early schizogony and is later processed into the 65-kDa antigen in segmenting schizonts. In Nonidet P-40, these antigens are non-covalently associated with two other proteins of 40 kDa and 42 kDa. The 40/42-kDa doublet is synthesised in parallel with the 82 kDa antigen and persists, apparently unchanged, till the end of the cell cycle.

Animals↗

PubMed1987

Effects of a selective alpha 1-adrenoceptor agonist, methoxamine, on sexual behavior and penile reflexes.

Methoxamine, an adrenergic agonist with selectivity for the alpha 1-adrenoceptor, when administered intraperitoneally 10 minutes prior to mating tests (1 to 5 mg/kg), effected reductions in the ejaculatory threshold, evidenced by a decrease in the number of intromissions preceding ejaculation. In mounting tests after penile anesthetization, a test which specifically assesses sexual motivation, 3 mg/kg methoxamine was without a stimulatory effect. Further, in penile reflex tests (ex copula) 1 mg/kg methoxamine was without effect, whereas 5 mg/kg decreased the number of erections, cups and flips per test, and increased the incidence of seminal emission. These data indicate a facilitation of the ejaculatory mechanism, both in and ex copula, coupled with an inhibition of erectile responses for moderate doses of methoxamine. Treatment of male rats with the alpha 2-adrenoceptor agonist clonidine (0.25 mg/kg, IP, five minutes pretest) drastically reduced the number of animals exhibiting intromissive and ejaculatory behavior in mating tests. This suppressive effect of clonidine was not prevented by prior treatment with methoxamine (3 mg/kg, 10 minutes pretest and five minutes preclonidine). Further, ST-91, a polar analog of clonidine which does not readily enter the central nervous system, was without effect on male sexual behavior. Since (1) the effects of methoxamine administration are not of similar quality or magnitude to those reported earlier after yohimbine, an alpha 2-adrenoceptor antagonist, (2) since concurrent stimulation of alpha 1- (by methoxamine) and alpha 2- (by clonidine) adrenoceptors is followed by a suppression of sexual behavior similar to that seen after clonidine alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

Neuropeptide Y (NPY)-induced feeding behavior in female rats: comparison with human NPY ([Met17]NPY), NPY analog ([norLeu4]NPY) and peptide YY.

Porcine neuropeptide Y (pNPY) administered into the third ventricle of the brain is known to elicit a powerful feeding response in steroid-treated ovariectomized and intact male rats. The present study compared the effects of pNPY and 3 structurally related peptides, human NPY (hNPY), an analog of NPY (NPY-A, [norLeu4]NPY) and peptide YY (PYY) on feeding behavior in intact female rats. Intraventricular administration of pNPY, hNPY, NPY-A and PYY over a dose range of 0.5 to 10 micrograms evoked feeding behavior to a varying extent. Cumulative food intake during 60 and 120 min was increased in a dose-related fashion at 0.5 and 2.0 microgram for the 4 peptides. Whereas the 10-micrograms dose of pNPY evoked a feeding response smaller than that seen after 2 micrograms, the responses to either 10 micrograms hNPY or 10 micrograms PYY were similar to that seen after 2 micrograms. The effects of these peptides on the time spent eating were quite different: while pNPY increased the time spent eating, this effect was not dose-related, whereas hNPY, NPY-A and PYY produced dose-related increments in the time spent eating. The most dramatic increment in local eating rate was observed after 2.0 micrograms pNPY, with lesser increments seen after 2.0 microgram hNPY and NPY-A. This increased local eating was apparently responsible for the highest cumulative food intake observed. These results demonstrate that (a) 2 micrograms pNPY is equally effective in stimulating feeding behavior in intact female rats as it is in steroid-primed ovariectomized female and intact male rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

PubMed1987

Topography of epitopes on a polymorphic schizont antigen of Plasmodium falciparum determined by the binding of monoclonal antibodies in a two-site radioimmunoassay.

The topographic distribution of common and variant epitopes on two divergent allelic forms of the 185-205K schizont glycoprotein of Plasmodium falciparum were studied by a two-site radioimmunoassay using monoclonal antibodies. Similarities in the conformation of the two molecules were apparent. On both antigens two distinct regions were mapped, each comprising of both strain-common and polymorphic epitopes. Epitopes common to the two PSAs were found to be closely associated with different variable epitopes in tertiary structure. It is suggested that this may contribute to parasite evasion of the host immune response.

Animals↗

PubMed1986

Failure of pimozide and metergoline to antagonize the RDS-127-induced facilitation of ejaculatory behavior.

Metergoline, a serotonin-receptor antagonist, when administered in either an ascorbic acid or ethanol containing vehicle was without effect on male rat copulatory behavior (3 mg/kg, 90 minutes pretest). When initially dissolved in several drops of acetic acid, however, the same dose of metergoline dramatically suppressed male rat sexual behavior. Thus, one-half of the treated rats failed to intromit and ejaculate, and those displaying the behaviors exhibited elongated intercopulatory and postejaculatory intervals. The administration of the putative dopamine-receptor agonist RDS-127 (2-N,N-di-n-propylamino-4,7-dimethoxyindane; 3 mg/kg, six minutes pretest) induced seminal emission ex copula and drastically reduced intromission frequency and ejaculation latency in copula, as well as effecting lesser reductions in the intercopulatory and postejaculatory intervals in two sequential copulatory series. RDS-127-induced seminal emission was effectively antagonized by pretreatment with the dopamine-receptor antagonist pimozide (250 micrograms/kg, two hours pretest), but not by pretreatment with metergoline. In contrast to seminal emission ex copula, pimozide pretreatment failed to antagonize the RDS-127 facilitation of ejaculatory behavior in copula. Metergoline pretreatment also failed to antagonize the RDS-127-induced facilitation of ejaculatory behavior in copula. However, RDS-127 prevented the suppressive effects of metergoline treatment, suggesting that RDS-127 has some agonistic action at serotonergic receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1986

Long-term weekly gonadal steroid treatment: effects on plasma prolactin, sexual behavior and hypothalamic-preoptic area catecholamines.

Treatment of intact female rats with weekly injections of estradiol benzoate followed 48 h later by progesterone reliably induced high levels of sexual behavior. After 15-20 treatments, 25% of one group of females became refractory to the sexual-activity-inducing effects of ovarian steroids. The apparent deficit in sexual behavior could not be attributed to variation in prolactin secretion, as long-term steroid treatment resulted in greatly elevated circulating prolactin levels (greater than 1 microgram/ml) which were equivalent in good sexual responders (lordosis quotient, LQ greater than or equal to 90) and sexually refractory females (LQ less than or equal to 20). In control rats, short-term steroid treatment (5 weeks) decreased dopamine (DA) and dihydroxyphenylacetic acid (Dopac) concentrations in the median eminence (ME) and induced good sexual behavior. Interestingly, a similar pattern of decreases in DA and Dopac levels of ME was observed in those long-term-treated rats displaying good sexual behavior but not in the sexually refractory females. Further, a significant increase in DA concentration in the preoptic-anterior hypothalamic area of the sexually refractory females was observed. These data are interpreted to suggest that severe and chronic elevations in circulating levels of prolactin, induced by chronic ovarian steroid treatment, are not universally associated with a disruption of sexual behavior and increased dopaminergic function in the ME, seen in females with normal sexual behavior, was conspicuously absent in female rats that became refractory to the sexual-behavior-inducing effects of ovarian steroids.

Animals↗

PubMed1986

Immunocytochemical differentiation of microtubules in the cytoskeleton of Giardia lamblia using monoclonal antibodies to alpha-tubulin and polyclonal antibodies to associated low molecular weight proteins.

In interphase trophozoites of Giardia lamblia, separate populations of microtubules constitute the four parts of the mastigont apparatus: flagella, ventral disc, funis and median body. Antigenic differences between the tubules have been investigated by light and electron immunocytochemistry after labelling with two monoclonal antibodies to alpha-tubulin (YL 1/2 and YOL 1/34 clones), and with polyclonal antibodies to Giardia tubule-associated proteins. Both anti-tubulins stained all tubules after isolated structures were fixed in formaldehyde, but different patterns of reactivity were shown by unfixed tubules. YL 1/2 antibodies labelled flagellar axonemes and basal bodies, funis and median body tubules. Disc microtubules were mostly unlabelled, but the antibody bound strongly to the outer edge of the disc where the ends of tubules are embedded. YOL 1/34 antibodies stained disc tubules uniformly, and cross-reacted with the median body but not with tubules of axonemes, basal bodies or funis. Antibodies to giardins 14A and 14B (approximately 30 000 Mr filament-forming proteins) localized these proteins in the microribbons attached to disc microtubules. The median body was also labelled by anti-giardins, indicating an ontogenetic relationship between this organelle and the ventral disc. A second set of approximately 30 000 Mr proteins with no immunoreactivity to anti-giardin was found in flagella purified without removing flagellar membranes. These polypeptides were Triton-soluble and therefore probably originated from an extra-axonemal site. A rabbit antiserum to the labile flagellar proteins specifically stained the two ventral flagella, but not the other six flagella on this cell.

Animals↗

PubMed1986

Plasma membrane isolated from Giardia lamblia: identification of membrane proteins.

Two methods are introduced for preparing plasma membranes from Giardia lamblia trophozoites. Isolated membranes were purified by centrifugation on either a sucrose step-gradient or a self-generated Percoll gradient, where they band at a density of approximately 1.04 g ml-1. In pure fractions, membranes formed vesicles or extensive sheets. Electron microscope profiles show that they are asymmetric with a thin filamentous coat on one side. Membrane proteins were resolved by SDS/PAGE. They included a major component of apparent Mr 75,000 (75 kDa), and additional bands detectable by gel staining at 58 kDa, 54 kDa, 32 to 38 kDa (5 bands), 22 kDa, and 15 to 20 kDa. To probe the surface location of proteins, gels were also prepared from Giardia cells that were surface radio-iodinated using the immobilised catalyst IODOGEN. The 75 kDa membrane protein was strongly labelled in the corresponding autoradiograph, also the bands at 58 kDa and 54 kDa, the 22 kDa polypeptide, and some faint bands not resolved in the isolated membrane preparations. The set of close-running bands at 32 to 38 kDa were not iodinated. The labelled 58 kDa and 54 kDa proteins comigrated with alpha and beta-tubulins. Controls showed that cytoskeleton and flagellar tubulins were not iodinated in this experiment, indicating that the labelled tubulin is surface-derived. The principal approximately 75 kDa surface protein identified in isolated membranes probably corresponds to an iodinatable and antibody-precipitated "82 kDa" antigen reported previously.

Animals↗

PubMed1985

Effects on penile reflexes and plasma hormones of hyperprolactinemia induced by MtTW15 tumors.

The effects of severe hyperprolactinemia induced by MtTW15 tumors (prolactin- and growth-hormone-secreting pituitary adenoma) on penile reflex activity and blood hormones were examined. There was no significant adverse effect of hyperprolactinemia on penile reflexes at 7, 14, or 21 days after tumor inoculation. However, a virtual elimination of penile reflex activity was observed 34 days after inoculation. Additionally, significant decrements in serum testosterone and dihydrotestosterone and an elevation in progesterone were seen at this time concomittant with greatly increased prolactin levels. The results suggest that erectile dysfunction may contribute to hyperprolactinemia-induced copulatory failure.

Animals↗

PubMed1985

Testosterone is not required for the enhancement of sexual motivation by yohimbine.

Yohimbine HCL (2 mg/kg, 20 min prior to testing) administration was followed by significant decreases in the latencies to initial mount, intromission and ejaculation in castrated male rats bearing 2 mm testosterone-containing Silastic capsules 51 days after castration. Further, yohimbine stimulated copulatory activity in castrated, nonhormone-treated male rats up to 91 days after castration. Finally, yohimbine induced mounting in intact, nonreceptive female rats. These observations indicate that testosterone is not required for the enhancement of sexual motivation by yohimbine and support the suggestion that alpha 2-adrenoceptors are involved in the modulation of sexual arousal.

Animals↗

PubMed1985

Central effects of RDS-127: sexual behavior after intracerebroventricular administration and in vitro receptor binding studies.

RDS-127, in a dose-related manner, induced seminal emission ex copula after intracerebroventricular (i.c.v.) administration. In mating tests initiated 6 min after i.c.v. administration, RDS-127 induced decreases in ejaculation latency and intromission frequency, with some rats ejaculating on the initial intromission. Additionally, penile reflexes were eliminated by 150 micrograms and 600 micrograms, but not by an intermediate dose. In in vitro radioligand binding studies, RDS-127 potently displaced [3H]DPAT binding to 5-HT1A sites in rat cortex (Ki = 14 +/- 4 nM) and was only moderately effective in displacing [3H]spiperone binding to dopaminergic D2 sites in rat striatum. RDS-127 was essentially ineffective at 5-HT1B sites labeled by [3H]5-HT in rat striatum (Ki = 13 000 +/- 4 000 nM). These data demonstrate that centrally administered RDS-127 mimics the previously reported alterations in sexual behavior after systemic treatment and that RDS-127 is a high affinity 5-HT1A agent with low affinity at the 5-HT1B binding site.

8-Hydroxy-2-(di-n-propylamino)tetralin↗