Changes in cholinergic and purinergic neurotransmission in pathologic bladder of chronic spinal rabbit

PURPOSE: We evaluated the changes in cholinergic and purinergic neurotransmission in pathologic bladder of chronic spinal rabbits. MATERIAL AND METHODS: Detrusor muscle strips were obtained from normal rabbits and chronic spinal rabbits with detrusor hyperreflexia and detrusor sphincter dyssynergia (DSD). Muscle strips were mounted in an organ bath, and transmural nerve electrical field stimulation (EFS: supamaximal voltage, 0.5 msec. duration, 10 second trains) was performed. The responses to EFS and agonists were determined by recording the isometric tension of muscle strips. RESULTS: Both normal and pathologic detrusor strips contracted in a frequency dependent fashion in response to transmural electrical nerve stimulation. At each frequency, atropine reduced the nerve-mediated contraction in a dose-dependent fashion and left an atropine-resistant response at a concentration of 1 microM. The atropine-resistant contraction was abolished by desensitization of P2X-purinoceptors with repeated exposure to alpha, beta-methylene ATP (10 microM). The atropine sensitive (cholinergic) and resistant (purinergic) contractions increased with an increase in frequency and reached maximum at 20 Hz. The relative contribution of cholinergic and purinergic transmission to the nerve-mediated contraction was determined at this frequency. In normal detrusor, the cholinergic and purinergic components were approximately 40% and 60%. In pathologic detrusor, the cholinergic component increased to 75% whereas the purinergic component decreased to 25%. Exogenously administered acetylcholine and ATP produced dose-dependent contractions of detrusor strips. The concentration-response curves for each agonist did not show significant differences between normal and pathologic detrusor. CONCLUSION: These results suggest that neurotransmission is shifted to a cholinergic dominance in pathologic rabbit bladder affected by detrusor hyperreflexia and DSD.     

Roles of histamine receptor in a guinea pig asthma model

A case of parotidomegaly associated with anorexia is described, and the psychoneurological and etiopathogenetic theories of main feed disorders (anorexia/bulimia) are discussed. Particularly, a rare pathology of maxillofacial interest is presented: parotidomegaly associated with anorexia. The diagnosis and differential diagnosis of this pathology is discussed.     

Neurokinin induced inositol phosphate production in guinea pig bladder

PURPOSE: To examine second messenger pathways involved in neurokinin induced bladder contractions. MATERIALS AND METHODS: Neurokinin induced changes in inositol phosphate production and in adenylyl cyclase activity are measured in the guinea pig bladder. RESULTS: Substance P, substance P methyl ester, neurokinin A, and neurokinin B each increase [3H]-inositol phosphate production in the guinea pig bladder. Substance P (10(-6) M) increases [3H]-inositol trisphosphate levels within 30 sec. Substance P and neurokinin A have an additive effect on inositol phosphate production, however substance P (10(-5) M) or neurokinin A (10(-5) M) induced inositol phosphate production is less than that induced by carbachol (10(-5) M). Neurokinin B and to a lesser extent neurokinin A inhibit forskolin-activated adenylyl cyclase activity. CONCLUSIONS: These data are compatible with neurokinin-induced inositol phosphate production being coupled to increases in contractile force of the guinea pig urinary bladder, however more than one second messenger pathway may be involved.     

Colocalization of nitric oxide synthase and some neurotransmitters in the intramural ganglia of the guinea pig urinary bladder

The distribution of nitrergic neurons was investigated by using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and nitric oxide synthase (NOS) immunohistochemistry in wholemount preparations of the urinary bladder in guinea pigs. Both NADPH-d+ and NOS+ neurons were located predominantly in the bladder base. Double staining showed that 70.9% of the NADPH-d+ neurons coexpressed NOS. Acetylcholinesterase histochemistry revealed that a majority of the intramural neurons were reactive, and about half of them (51.4%) were double labelled for NOS. Tyrosine hydroxylase-positive neurons were also distributed mainly in the bladder base but in a neuronal population that was separate from the preponderant NADPH-d+ neurons. Vasoactive intestinal polypeptide immunoreactivity was also detected in the some of intramural ganglion cells, in which 21.3% of them coexpressed NADPH-d. Calcitonin gene-related peptide and substance P immunoreactivities were confined to nerve fibers, often in close association with NADPH-d+ cells or extended along the blood vessels. These results have demonstrated the colocalization of NADPH-d and NOS in the majority of intramural ganglion cells. Many of the nitrergic neurons are apparently cholinergic, indicating that they are parasympathetic postganglionic neurons, and this underscores NO as the major neuromodulator in the parasympathetic nerves in the bladder walls. The localization of vasoactive intestinal polypeptide in nitrergic neurons suggests that the peptide may complement NO for regulation of micturition reflex. The close relationship of NADPH-d-reactive intramural neurons with calcitonin gene-related peptide and substance P fibers, most probably derived from dorsal root ganglion cells, suggests that NO released from the local neurons may exert its influence on the sensory neural pathways in the urinary bladder.     

Activation of dopaminergic and cholinergic neurotransmission by tachykinin NK3 receptor stimulation: an in vivo microdialysis approach in guinea pig

The regulation of dopaminergic and cholinergic function by neurokinin-3 (NK3) receptor activation was examined in vivo in urethane-anaesthetized guinea pigs with microdialysis probes. The local application of the NK3 tachykinin receptor agonist senktide in the region of dopamine cell bodies (pars compacta of the substantia nigra and ventral tegmental area) and in the area of cholinergic cell bodies (septal area) markedly enhanced the extracellular dopamine (DA) and acetylcholine (ACh) concentration throughout their respective target areas, i.e. striatum, nucleus accumbens, prefrontal cortex for dopaminergic systems and hippocampus for cholinergic neurons. The enhancing effect of senktide on neurotransmitter release was dose dependently blocked by the selective non-peptide NK3 receptor antagonist SR142801 (0.1-1 mg/kg, i.p.), whereas its inactive S-enantiomer SR142806 (0.3-1 mg/kg, i.p.) did not exert any antagonistic activity on the effect of intranigral or intraseptal application of senktide. These results demonstrate that NK3 receptors can modulate the activity of central DA and ACh systems.     

H1- and H2-receptors for histamine in the ileum of the guinea pig

Clark's equation used in its inverted form (double reciprocal plots) was shown to be valid regarding the interaction of histamine with its receptors in the guinea-pig ileum. Burimamide, a typical H2-receptor antagonist potentiated the stimulating effect of small concentrations of histamine (10(-6) M) on the intestinal smooth muscle, probably, by blocking H2-receptors which may show the opposite effect. The calculated parameters of affinity (pKn) remained constant under different conditions. A linear correlation between length of the preparation and maximal contractions has been found.     

The action of histamine on the slow-wave and spike activities of the cat and guinea pig ureter

Histamine increased the oscillation frequency of the slow-wave activity in the cat ureter and changed the characteristics of the AP plateau phase. A specific and sensitive to histamine electrogenic Na/Ca agent seems to take part in generation of the ureter spontaneous activity.     

Betadrenoceptors in urinary bladder

When rat peroneal nerves were fixed under 5 different fixation schedules with various osmolalities (310, 410 and 900 milliosmoles), the frequency of abnormality of mitochondria in the axoplasm was lowest in myelinated fibers fixed with 6% glutaraldehyde in 0.1 M cacodylate buffer (900 milliosmoles). The relationship of axonal area to number of myelin lamellae was significantly different between high-osmolality fixation and low-osmolality fixation. Assuming that the number of myelin lamellae remains the same with fixation, there was a significant shrinkage in the axonal area of fibers with high-osmolality fixation. The amount of shrinkage was less in larger fibers than in smaller fibers fixed at high osmolality.     

Propagation by sporulation in the guinea pig symbiont Metabacterium polyspora

The Gram-positive bacterium Metabacterium polyspora is an uncultivated symbiont of the guinea pig gastrointestinal tract. Here we present evidence that in M. polyspora vegetative cell division has taken on a minor, and apparently dispensable, role in propagation. Instead, this unusual bacterium has evolved the capacity to produce progeny in the form of multiple endospores. Endospore formation is coordinated with transit of the bacterium through the gastrointestinal tract of the guinea pig. For the majority of cells, sporulation is initiated in the ileum, whereas later stages of development take place in the cecum. We show that multiple endospores are generated both by asymmetric division at both poles of the cell and by symmetric division of the endospores at an early stage of their development. Our findings suggest that M. polyspora represents an intermediate step in the evolution of a novel mode of cellular propagation that originates with endospore-forming Bacillus and Clostridium spp., which reproduce by binary fission, and extends to Epulopiscium spp., which create multiple viviparous offspring by a process of internal reproduction.     

Block of potassium currents in guinea pig ventricular myocytes and lengthening of cardiac repolarization in man by the histamine H1 receptor antagonist diphenhydramine

Treatment with second generation histamine H1 receptor antagonists has been associated with lengthening of the Q-T interval and proarrhythmia. Similarly, lengthening of the Q-T interval has been reported in patients after overdosing with diphenhydramine (DPH), a first generation agent. Therefore, our study was designed 1) to assess effects of DPH on cardiac repolarization and 2) to characterize effects of the drug on major voltage-dependent cardiac K+ currents. First, we noticed that oral administration of DPH at usual dosages to healthy volunteers or to patients (prior to angioplasty) was associated with prolongation of the Q-Tc interval. Although this effect was modest in most individuals, Q-Tc was increased more than 20 ms in 7 of 20 patients. Second, we noticed that exposure of isolated guinea pig hearts to DPH 10(-5) M caused a lengthening of monophasic action potential duration. This effect was potentiated by the combined perfusion of other K+ channel blockers such as indapamide. Finally, experiments performed with the patch-clamp technique demonstrated unequivocal block of the rapid component of the delayed rectifier (IKr) by DPH; however, IC50 determined for block of IKr (3 x 10(-5) M) is approximately 40-fold greater than plasma concentrations of the drug measured at usual dosages (7 x 10(-7) M). Consequently, in agreement with the long-term clinical use of the drug, prolongation of cardiac repolarization should be minimal in most patients at usual dosages but may be observed with overdosing. Nevertheless, caution remains since excessive lengthening of cardiac repolarization may occur after administration of DPH with other drugs due to 1) concomitant block of other ionic currents or 2) pharmacokinetic interactions leading to toxic concentrations of DPH.     

Desulphation of heparin by mice and guinea pig leukocytes

A series of 603 patients referred with atypical Papanicolaou smears was evaluated by repeat smears, colposcopically directed cervical biopsies, and endocervical curettage. These techniques as a unit can establish an accurate outpatient diagnosis superior to any of these modalities used alone and comparable with findings in conization and hysterectomy specimens. Endocervical curettage has made a unique contribution to the evaluation of such patients; these curettings have allowed examination of tissue fragments and are more reliable in diagnosing neoplasia than are endocervical smears. Invasive carcinoma and its precursors confined to the anatomic endocervical canal can be recognized by this technique, and conversely the absence of neoplastic epithelium in adequate endocervical curettings rules out occult carcinoma. Indications for conization of the cervix are discussed in reference to the other biopsy and cytologic findings, and guidelines are presented for patient management, stressing clinicopathologic correlation and cooperation.     

Direct evidence for histamine H3 receptor-mediated inhibition of norepinephrine release from sympathetic terminals of guinea pig myocardium

AIM: To study the histamine H3 receptors mediated inhibition of norepinephrine (NE) release from cardiac sympathetic terminals of guinea pig isolated atria. METHODS: Release of NE induced by electric field stimulation (50 mA, 5 ms) in the bath solution was measured by HPLC-ECD. RESULTS: The release of NE caused by field stimulation was attenuated by (R)-alpha-methyl-histamine (alpha-MeHA, 0.1 nmol.L-1(-10) mumol.L-1) in a concentration-dependent manner. Thioperamide concentration-dependently antagonized the inhibition of alpha-MeHA. Blockade of H1, H2, alpha 2, beta 2-receptors failed to prevent the inhibitory effect of alpha-MeHA. Thioperamide (1 nmol.L-1(-10) mumol.L-1), when used alone, concentration-dependently facilitated the release of NE evoked by field stimulation. CONCLUSION: The presynaptic histamine H3-receptors inhibited the NE release from cardiac sympathetic terminals.     

Effect of short and long-term exposure to diesel exhaust on sensitivity of guinea pig tracheal preparation to histamine

Single exposure, to diesel exhaust (1 part exhaust diluted by 5 parts of clean air) reduced EC50 of histamine indicating hyperresponsiveness of the receptors in trachea of exposed guinea pigs. In contrast, following repeated exposure for 7, 14 or 21 days (15 min/day), EC50 was progressively increased indicating the possibility of down-regulated histamine receptors. Further, simultaneous significant increase in histamine levels (bioassayed on guinea pig ileum) in bronchial airway lavage fluid supports the aforementioned hypothesis. The change in lung/body weight ratio and suspended particulate matter deposited on filters followed the same temporal pattern as EC50. The findings are suggestive of differential effects of diesel exhaust on airway depending upon the duration of exposure.     

Inhibition in the human urinary bladder by gamma-amino-butyric acid

OBJECTIVE: To investigate the effects of gamma-amino-butyric acid (GABA) on detrusor activity in man to determine whether it has any inhibitory effect on detrusor contraction. The inhibitory neurotransmitter GABA has been found in mammalian urinary bladders and the effects of GABA on detrusor activity in the rabbit bladder has previously been described [1]. MATERIALS AND METHODS: Human detrusor muscle strips, obtained at cystectomy, were made to contract by electrical stimulation of their autonomic nerves or by the addition of carbachol in a superfusion apparatus. GABA and its analogues were added to the superfusion chamber and any changes in the responses were measured. RESULTS: The electrically evoked nerve-mediated contractions in human bladder muscle were exclusively cholinergic. GABA inhibited nerve-mediated contractions in human detrusor muscle-strips by the activation of the GABAB receptor, since baclofen (a GABAB receptor agonist) produced similar inhibition and muscimol (a GABAA receptor agonist) did not. There was no inhibition of carbachol-mediated contractions by GABA. CONCLUSION: This in vitro study shows that GABA has a peripherally mediated inhibitory effect on excitatory neurotransmission in human detrusor muscle. The site of action is on the post-ganglionic nerves and appears to be mediated via the GABAB receptor.     

Active secretion of hypoxanthine and xanthine by guinea pig jejunum in vitro

Isolated epithelium of guinea pig jejunum secretes hypoxanthine and xanthine by a transport process that is capable of uphill transport and dependent on metabolic energy supply. Unidirectional influx of hypoxanthine across both the luminal and the contraluminal cell membrane appears to be saturable; influx across the contraluminal membrane is inhibited by 2,4-dinitrophenol (DNP). Efflux across the luminal membrane is diminished by DNP; efflux across the contraluminal membrane is increased by DNP. This evidence suggests the existence of a mediated transport system both in the luminal and the contraluminal cell membrane. Additionally, intracellular metabolism of hypoxanthine seems to regulate transepithelial permeation: increased hypoxanthine salvage by the phosphoribosyltransferase reduces the rate of secretion. However, the incorporation of hypoxanthine into the nucleotides is limited when the hypoxanthine is added to the luminal side of the epithelium, and the permeation rate in the absorptive direction is not markedly influenced by the rate of hypoxanthine salvage. These findings are a further example of the functional orientation of the jejunal epithelial cells with respect to enzymic activity and transepithelial transport properties.     

Signal transduction pathways in guinea pig sperm

Trifluoperazine (TFP), the antagonist of calmodulin (CaM), significantly stimulated the capacitation and acrosome reaction of guinea pig spermatozoa at the concentration of 10-100 mumol/L, independent of the external Ca2+. Forskolin, dbcAMP and caffeine evidently promoted the occurrence of acrosome reaction of spermatozoa at early capacitation stage (5 h) in nonsynchronous system but not in synchronous system. If the spermatozoa were capacitated for 15 h in synchronous system, the above three drugs significantly stimulated acrosome reaction in a Ca(2+)-independent manner. Protein kinase C activators, i.e. phorbol 12-myristate 13-acetate (PMA) and phorbol 12,13-dibutyrate (PDB) did not influence the occurrence of acrosome reaction of spermatozoa at early capacitation stage, but significantly increased the acrosome reaction rate in capacitated spermatozoa in a Ca(2+)-independent manner. In contrast, PKC inhibitor staurosporine significantly inhibited the occurrence of acrosome reaction.