The effect of reserpine, chlorpromazine and neumbutal on levels of dipeptides in rat brain and muscle

Rat brain levels of histidine, carnosine, and homocarnosine were determined after intraperitoneal injection of chlorpromazine (CPZ), sodium pentobarbital (PB), or reserpine (RSP). At the same time, rat muscle levels of histidine, carnosine, and anserine were determined. RSP, CPZ, and PB significantly lowered brain homocarnosine levels and RSP raised histidine levels. RSP, CPZ, and PB significantly lowered levels of muscle carnosine and anserine. PB and CPZ also lowered levels of muscle histidine.     

Antagonistic response of chlorpromazine and formalin in effecting quantitative changes in the neurosecretory material in the teleost Clarias batrachus (L.).

The hypothalamoneurohypophysial complex of Clarias batrachus maintains an appreciable quantity of neurosecretory material (NSM) under normal conditions. Stress caused by the injection of formalin depletes 70-90% of the stainable NSM from all the component parts of the neurosecretory system, namely the nucleus preopticus (NPO) the neurosecretory tract, and the neurohypophysis. Formalin apparently releases the NSM and stimulates the neurons of the NPO. Chlorpromazine (CPZ) treatment seems to cause quantitative increase of NSM throughout the neurosecretory complex. Simultaneous injection of CPZ and formalin showed that the formalin-induced depletion of NSM could be inhibited by CPZ.     

Effects of chlorpromazine on pattern and flash ERGs and VEPs compared to oxazepam and to placebo in normal subjects

Antidopaminergic drugs delay the pattern-reversal VEP (P-VEP) and the flash VEP (F-VEP) and, in separate studies, reductions in the amplitude and increases in the latencies of scotopic ERGs have been reported. This study investigated the effects of chlorpromazine (CPZ) on the pattern ERG (P-ERG), P-VEP, flash ERGs and VEPs and oscillatory potentials (OPs). Normal volunteers (N = 15) were administered a placebo, or a single dose of CPZ 100 mg or oxazepam (OZP) 15 mg at weekly intervals, in a double-blind crossover design. A gold foil-ipsilateral ear derivation and an Oz′-Fz derivation were used for the ERG and VEP recordings, respectively. The latencies of ‘mixed’ and cone ERGs were significantly prolonged after CPZ compared to both placebo and to OZP. Amplitudes of rod- and cone-dominated ERGs were reduced following CPZ administration. All components of the OPs were significantly delayed after CPZ administration. No significant intertreatment differences were found in the F-VEP results. The P-ERG P50 peak and the P-VEP N70 and P100 peaks were significantly delayed after CPZ in the case of 28′ checks but not 55′ checks. Retinocortical times and P-ERG and P-VEP amplitudes were not significantly affected. In contrast to CPZ, the administration of OZP had virtually no significant effects compared to placebo. These findings suggest that the antidopaminergic CPZ has a primary effect on retinal electrophysiology. Similar findings have been reported in Parkinson's disease and in animal models.     

氯丙嗪在斑马鱼胚胎和早期幼鱼发育过程中的神经毒性作用

目的 采用模式动物斑马鱼作为研究对象,观察氯丙嗪（chlorpromazine,CPZ）暴露对胚胎和幼鱼早期神经发育的影响.方法 在一般毒性评价的基础上,通过整体胚胎细胞凋亡检测和脑组织病理学检查,了解CPZ对神经发育的器质性改变;采用神经行为学方法,包括幼鱼触动逃避反应、自发运动以及惊恐逃避反射等,研究氯丙嗪暴露所致的神经发育功能性障碍.结果斑马鱼胚胎受精后6 h（6 hpf）～72 hpf暴露于CPZ（≥5 mg/L）可引起胚胎和幼鱼死亡、致畸和幼鱼孵化延迟,并呈浓度和时间依赖性;采用吖啶橙染色检测36 hpf整体胚胎凋亡细胞,发现凋亡细胞主要集中在胚胎中脑、后脑、丘脑以及中后脑连接区、脊索和尾部等处;脑组织病理学检测发现,7dpf幼鱼颅腔增大、脑体积减小、脑细胞缩小且细胞间隙增宽.6～72 hpf CPZ（≥0.0625 mg/L）暴露后,幼鱼神经行为学研究发现,CPZ（≥0.125 mg/L）可引起3dpf幼鱼触觉运动能力下降;CPZ（≥0 5 mg/L）可浓度依赖性地抑制幼鱼自发运动,并出现僵直不动、震颤或快速刻板式转圈运动等行为改变;光惊恐实验中,暗环境下各暴露组幼鱼对突发强光刺激均表现为惊跳逃避,并且暗-光交替期运动加速度变化与对照组无显著差异;在撤除光源后,1mg/L和2 mg/L暴露组幼鱼暗适应时程缩短,而0.125 mg/L和0.25 mg/L暴露组暗适应时程延长,提示CPZ对外界刺激引发的幼鱼活跃游动有抑制和促进双重毒性作用.结论 CPZ暴露对斑马鱼胚胎和幼鱼具有明显的神经发育毒性作用.模式动物斑马鱼作为一种高通量筛选模型在外源性化合物神经发育毒性评价中具有较好的应用前景.     

Changes of serum and tissue amoxicillin levels following chlorpromazine administration in rats

Patients under treatment with chlorpromazine (CPZ) may need antibacterial administration. Amoxicillin is the most widely used antibacterial agent in dental infections. The aim of this study was to investigate the possible interactions between amoxicillin and CPZ and the eventual changes in amoxicillin levels using an experimental model in rats. Thirty male Wistar rats weighing 200-260 g were used for this study. They were divided into three groups: group A taking 15 mg/kg amoxicillin every 8 h for 4 days; group B taking amoxicillin as group A, and 1.07 mg/kg CPZ every 8 h for 4 days; and group C taking only CPZ in the same dose as group B. Drugs were given per os for 4 days, using a special catheter. The rats were sacrificed 2 h after the last dose. Antibacterial levels were estimated in blood serum, jaw bones, tongue and liver. An increase in the level of amoxicillin in serum as well as in tissues was observed in group B. No significant antibacterial activity of CPZ was detected. The increase in amoxicillin concentration in serum and jaws of group B was statistically significant in relation to group A.     

Responses of the neurosecretory cells of the freshwater snail,Indoplanorbis exustus to hydrothermal stress

Changes in the neurosecretory cell cytology of I. exustus subjected to hypertonic saline (0.1 ml of 1.5%/snail) loading and thermal stress (35°C) for two hours, have been investigated. Of the two types of neurosecretory cells A and B that are present in the central nervous system (CNS) of I. exustus, striking changes were evident only in B cells. After both treatments, there was about 33% decline in NSM (Neurosecretory material) intensity. However, the nuclear diameter of B cells was significantly (P < 0.001) increased in the snails administrated with hypertonic saline unlike in those exposed to 35°C wherein significant (P < 0.005) decline was evident. The adaptive significance of the neuroendocrine system of I exustus is discussed in relation to hydrothermal stress.     

Chlorpromazine protects against apoptosis induced by exogenous stimuli in the developing rat brain

Background

Chlorpromazine (CPZ), a commonly used antipsychotic drug, was found to play a neuroprotective role in various models of toxicity. However, whether CPZ has the potential to affect brain apoptosis in vivo is still unknown. The purpose of this study was to investigate the potential effect of CPZ on the apoptosis induced by exogenous stimuli.

Methodology

The ethanol treated infant rat was utilized as a valid apoptotic model, which is commonly used and could trigger robust apoptosis in brain tissue. Prior to the induction of apoptosis by subcutaneous injection of ethanol, 7-day-old rats were treated with CPZ at several doses (5 mg/kg, 10 mg/kg and 20 mg/kg) by intraperitoneal injection. Apoptotic cells in the brain were measured using TUNEL analysis, and the levels of cleaved caspase-3, cytochrome c, the pro-apoptotic factor Bax and the anti-apoptotic factor Bcl-2 were assessed by immunostaining or western blot.

Findings

Compared to the group injected with ethanol only, the brains of the CPZ-pretreated rats had fewer apoptotic cells, lower expression of cleaved caspase-3, cytochrome c and Bax, and higher expression of Bcl-2. These results demonstrate that CPZ could prevent apoptosis in the brain by regulating the mitochondrial pathway.

Conclusions

CPZ exerts an inhibitory effect on apoptosis induced by ethanol in the rat brain, intimating that it may offer a means of protecting nerve cells from apoptosis induced by exogenous stimuli.     

Human cells and cell membrane molecular models are affected in vitro by chlorpromazine

This study presents evidence that chlorpromazine (CPZ) affects human cells and cell membrane molecular models. Human SH-SY5Y neuroblastoma cells incubated with 0.1 mM CPZ suffered a decrease of cell viability. On the other hand, phase contrast microscopy observations of human erythrocytes indicated that they underwent a morphological alteration as 1 μM CPZ changed their discoid normal shape to stomatocytes, and to hemolysis with 1 mM CPZ. X-ray diffraction experiments performed on dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE) bilayers, classes of the major phospholipids present in the outer and inner sides of the erythrocyte membrane, respectively showed that CPZ disordered the polar head and acyl chain regions of both DMPC and DMPE, where these interactions were stronger with DMPC bilayers. Fluorescence spectroscopy on DMPC LUV at 18 °C confirmed these results. In fact, the assays showed that CPZ induced a significant reduction of their generalized polarization (GP) and anisotropy (r) values, indicative of enhanced disorder at the polar head and acyl chain regions of the DMPC lipid bilayer.     

Effect of Chlorpromazine on Human and Murine Intracellular Carboxylesterases

Clinical use of chlorpromazine (CPZ), an antipsychotic drug, is limited due to its hepatotoxicity. CPZ is found to inhibit in vitro intracellular carboxylesterases (CE), such as alpha-naphthyl acetate esterase, naphthol AS-D chloroacetate esterase, and alpha-naphthyl butyrate esterase in polymorphonuclear neutrophils, hepatocytes, and neuronal brain cells from mice. CPZ inhibits CE of all these cell types, whereby the degree of the inhibition depends on the incubation time and CPZ concentration. The polymorphonuclear neutrophils are most sensitive to CPZ. Comparable results were obtained with polymorphonuclear neutrophils from mice and humans. Since leukocytes are much more available than hepatocytes or neuronal cells in humans, we assume that CE in peripheral blood leukocytes (neutrophils and monocytes) can be used as markers for indication of pending liver damage by CPZ.     

Interaction of chlorpromazine with the human erythrocyte membrane

The interaction of the amphipath chlorpromazine (CPZ) with the human erythrocyte membrane was evaluated. The partition coefficient of CPZ between the membrane bilayer and the aqueous compartment, measured spectrophotometrically, ranged between 1 and 3 X 10(3). An independent estimate, 4.6 X 10(3), was obtained by a novel method which avoided the measurement of binding and determined instead the variation of the hemolytic potency of the amphipath with the ratio of buffer volume to membrane volume. The maximal uptake of CPZ exceeded 2 X 10(9) molecules/red cell, corresponding to a volume greater than that of the bilayer itself. Such heavily loaded membranes were increased in thickness more than 2-fold, suggesting the formation of a CPZ-rich zone at the center of the bilayer. Ghosts loaded with massive levels of CPZ condensed approximately 20-fold in surface area and increased proportionately in thickness, suggesting the formation of a novel CPZ-lipid solution. CPZ caused hemolysis by a colloid-osmotic mechanism. By measuring the simultaneous uptake of mannitol and sucrose, we determined that CPZ induced holes of constant size but variable number. If circular, the holes would have had a diameter of approximately 14 A. The time-averaged number of holes ranged from 0.09 per cell (signifying intermittency) to 16. Freeze-fracture electron microscopy of CPZ-treated red cells revealed multiple round patches of nearly particle-free bilayer up to 0.3 micron in diameter with crowding of the intramembrane particles into the surrounding membrane. We interpret these images to signify lateral phase separation within the CPZ-treated bilayer. Hemolysis could, therefore, result from the intermittent opening of weak seams at phase boundaries; these could then be fluctuating slits approximately 14 A in width and of variable length, rather than simple circular holes.     

Subcellular localization and function of mouse radial spoke protein 3 in mammalian cells and central nervous system

Radial spoke protein 3 (RSP3) was first identified in Chlamydomonas as a component of the radial spoke. The mammalian homologue of the Chlamydomonas RSP3 gene is mainly expressed in testis and developing central nervous system (CNS). However, the subcellular localization and function of mammalian RSP3 in the developing brain and mammalian cells remain poorly understood. Here we show that the mouse RSP3 accumulates at the perinuclear region of Chinese hamster ovary (CHO) and 293T cells. Detailed analysis shows that the mouse RSP3 is not co-localized with the endoplasmic reticulum or Golgi apparatus markers in CHO cells. Using in utero electroporation, we found that over-expression of mammalian RSP3 increases the percentage of neurons reaching the upper cortical plate. In vivo analysis shows that the mouse RSP3 mainly accumulates in the proximal cytoplasmic dilation of the leading process of the migrating cortical neurons. Furthermore, we find that the mammalian RSP3 concentrates in the ependymal cilia as a component of the cilia. Thus, our data provide the first evidence for the subcellular localization and function of mammalian RSP3 in mammalian cells and developing CNS.     

External ATP-induced passive permeability change and cell lysis of cultured transformed cells: action in serum-containing growth media

External ATP causes a marked increase in the passive permeability to phosphorylated metabolites in several types of transformed cells in alkaline medium containing low concentrations of Ca2+, but not in untransformed cells. Such increased membrane permeability with external ATP was also observed in B16 melanoma cells at pH 7.4-7.5 in both Tris-buffered saline and a growth medium containing 10% calf serum and divalent ions at normal concentrations, although a higher concentration of ATP was required. The permeability change in the growth medium was significantly enhanced by calmodulin-interacting drugs, such as trifluoperazine (TFP), N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W7) and chlorpromazine (CPZ). As expected, prolonged exposure of the cells to ATP in the serum-containing medium led to cell lysis. This ATP-dependent cell lysis was observed only in several transformed cell lines, and not in untransformed mouse fibroblasts. These results indicate that the effect of ATP on the membrane permeability in transformed cells is elicited under the physiological conditions and this would be useful in some limited way for cancer chemotherapy management.     

Selective amphipathic nature of chlorpromazine binding to plasma membrane bilayers

Chlorpromazine (CPZ), an antipsychotic agent shown to inhibit the action of various neurophysiological receptors, also exhibits preferential association with the plasma membrane, inducing stomatocytic morphological response in red blood cells (RBC). Given the cationic nature of CPZ, fluorimetry, pH titration, and red cell morphological studies were performed to assess the associative predilection of CPZ for anionic membrane components. CPZ fluorescence intensity increased 320-370% upon addition of phosphatidylcholine (PC) small unilamellar vesicles (SUVs) to aqueous CPZ, indicating an affinity of the drug for lipidic phases. After removal of unbound drug, CPZ fluorescence increased up to 92% with increasing phosphatidylserine (PS) in the lipid phase (up to 30 mol% of total lipid), suggesting a preferential association of the drug with anionic lipids. In studies of pH titration, the pK(a) of CPZ in the presence of Triton X-100 micelles or phospholipid SUVs increased with increasing anionicity of the lipidic phase [7.8 with Triton X-100, 8.0 with PC, 8.3 with phosphatidylglycerol (PG)], lending further support to preferential drug interaction with anionic lipidic components. At 0 degrees C, CPZ-induced red cell shape change was less extensive in cells made echinocytic by adenosine triphosphate (ATP) depletion, compared to cells made echinocytic by PS treatment following vanadate preincubation. This suggests that polyphosphoinositide lipids are CPZ membrane binding sites. Since polyphosphoinositide lipids are implicated as important intermediates in a number of receptor-mediated cell signaling pathways, evidence of association with these specific lipids provides a means by which psychoactive drugs may induce neurophysiological effects through direct interaction with general membrane components.     

Xenopus radial spoke protein 3 gene is expressed in the multiciliated cells of epidermis and otic vesicles and sequentially in the nephrostomes

We describe the phylogenetic analysis and expression pattern of the Xenopus radial spoke protein 3 (RSP3) gene during early development. The Xenopus RSP3 protein presents characteristic features of the RSP3 family. It contains a radial spoke domain, which is 75 and 72 % identical to the corresponding region of human and Chlamydomonas RSP3 proteins, respectively. Examination of the phylogenetic relationship between the Xenopus RSP3 protein and its known homologues from different deuterostomes indicates that the RSP3 proteins are highly conserved among deuterostomes. Whole-mount in situ hybridization analyses show that Xenopus RSP3 is a maternal mRNA enriched in the animal hemisphere during cleavage stages. The expression is detected in the dorsal region of the embryo during gastrulation, then in the presumptive neuroectoderm at the end of gastrulation. During neurulation and at the subsequent stages, the expression of RSP3 mRNA is detected in the entire multiciliated cells of epidermis. At tail-bud stages, it is progressively expressed in the otic vesicles and sequentially expressed in the nephrostomes. Expression could be also detected in the floor plate of the neural tube. This expression pattern persists until at least late tail-bud stages.     

Differential binding of chlorpromazine to human blood cells: Application of the hygroscopic desorption method

Using a modified hygroscopic desorption method (HDM) the binding of chlorpromazine (CPZ) to human blood cells was investigated in the concentration range from 0.01 to 100 μmol/1. For erythrocytes and ghosts the ratio between cell bound and free drug concentration was constant up to 60 μmol/1 CPZ. Saturable binding, however, was observed for lymphocytes, granulocytes and less pronounced for platelets. In contrast to red cells, CPZ binding to white cells and platelets was strongly dependent on pH. For all blood cells a sharp decrease in binding occurred at drug concentrations higher than 60 μmol/1. This can hardly represent a true saturation of binding sites, since membrane damaging effects occur at these concentrations. Our results suggest that binding of CPZ to erythrocytes represents an interaction at the water-membrane interphase. For the different binding pattern of white cells, the cell organelles, the cytoplasma and the different composition of the membranes might be of importance.     

The evolution of neurosecretory centers in bilaterian forebrains: insights from protostomes

Forebrain neurosecretory systems are widespread in the animal kingdom. This review focuses on recent molecular data from protostomes, discusses the original complexity of the bilaterian forebrain neurosecretory system, provides an evolutionary scenario for the emergence of the vertebrate preoptic area/hypothalamus/neurohypophysis and suggests a possible function for an ancient set of sensory-neurosecretory cells present in the medial neurosecretory bilaterian forebrain.     

In vitro andin vivo effect of chloropromazine,imipramine and lithium chloride on monoamine oxidase activity in rat brain mitochondria

Chloropromazine (CPZ) and imipramine at a concentration of 1×10^–3 M inhibit rat brain mitochondrial monoamine oxidase activity in vitro by 70 and 55% respectively, while lithium, even at a concentration of 0.05 M, inhibits the activity of this enzyme very negligibly (4%). In vivo, these drugs at a dose level of 56 mg CPZ, 76 mg Jimipramine and 76 mg lithium chloride/Kg body wt., did not cause any observable variation from normal in brain mitochondrial monoamine oxidase activity.To whom correspondence should be addressed.     

Immunohistochemical localization of carboxypeptidases D, E, and Z in pituitary adenomas and normal human pituitary.

Carboxypeptidases may play important role(s) in prohormone processing in normal and neoplastic adenohypophyseal cells of the pituitary. We have recently demonstrated carboxypeptidase E (CPE) and carboxypeptidase Z (CPZ) in the majority of adenohypophyseal cells with carboxypeptidase D (CPD) immunoreactivity largely confined to adrenocorticotrophs. This study evaluated the expression patterns of CPE, CPD, and CPZ immunoreactivity in 48 pituitary adenomas. Our immunohistochemistry demonstrated extensive intracytoplasmic immunoreactivity for CPE, CPD, and CPZ in adrenocorticotrophic hormone (ACTH)-producing adrenocorticotroph cells, prolactin-producing lactotroph cells, and growth hormone (GH)-producing somatotroph cell adenomas, all of which require carboxypeptide processing of prohormones to produce active endocrine hormones. In contrast to the restricted expression in the normal adenohypophysis, CPD appeared to be widespread in the majority of adenomas, suggesting that CPD levels are increased in adenomas. In luteinizing hormone/follicle-stimulating hormone (LH/FSH)-producing gonadotroph adenomas, which do not require carboxypeptidases to produce gonadotropins, only CPZ immunostaining was demonstrated. In null-cell adenomas, CPE immunoreactivity was detected in the majority of tumors, but CPD and CPZ were identified only in a minority of cases. CPE in these cells may process other peptides critical for pituitary cell function, such as chromogranin A or B. These findings suggest that CPs participate in the functioning of pituitary adenomas.     

Chlorpromazine inhibits mitosis of mammalian cells

Chlorpromazine (CPZ) at minimally effective concentrations accumulates mammalian cells in mitosis without lethal effects on the cells. Star-metaphase morphology similar to effects seen with classical antimitotic compounds probably results from the preferential action of CPZ on a specific class of microtubules--the pole-to-pole microtubules of the mitotic spindle. At CPZ concentrations of 8 X 10(-6) M, flow cytometry indicates no effect of CPZ on the progress of cells through phases of the cell cycle other than mitosis (M). These results suggest a possible mechanism for toxic side effects of CPZ in man such as granulocytopenia and light sensitization.