Alterations in brain 5-hydroxytryptamine metabolism during the 'withdrawal' phase after chronic treatment with diazepam and bromazepam.

1 Daily administration of diazepam or bromazepam (10 mg/kg) for 22 days significantly increased the activity of mid-brain tryptophan hydroxylase by 36% and 39%, respectively. The concentration of tryptophan was also enhanced in the mid-brain region of rats subjected to benzodiazepine treatment.2 Chronic therapy with either of the two anti-anxiety agents enhanced the endogenous levels of 5-hydroxytryptamine and 5-hydroxyindoleacetic acid in cerebral cortex, hypothalamus, pons-medulla, mid-brain and striatum.3 Whereas diazepam treatment decreased (13%) the activity of monoamine oxidase in mid-brain, bromazepam failed to exert any effect, suggesting that the observed elevation in 5-hydroxy-indoleacetic acid levels is not associated with enhanced deamination of 5-hydroxytryptamine.4 Discontinuation of treatment for 48 h significantly decreased the activity of mid-brain tryptophan hydroxylase to levels that were significantly lower than those seen for benzodiazepine-treated and normal rats. The concentrations of mid-brain tryptophan and 5-hydroxytryptamine were also reduced in various brain regions examined.5 Withdrawal from diazepam or bromazepam therapy further augmented the levels of brain 5-hydroxyindoleacetic acid.6 The results demonstrate that the depressant effects on behaviour of these agents are accompanied by increased metabolism of 5-hydroxytryptamine in the brain. Withdrawal from these minor tranquillizers, on the other hand, reduces the synthesis of this indoleamine.

Alteration in biogenic amine levels in the snail Lymnaea acuminata by the latex of Euphorbia royleana.

The latex of Euphorbia royleana which has high molluscicidal and anti-cholinesterase activity against Lymnaea acuminata reduced the levels of 5-hydroxytryptamine (5HT) and dopamine (DA) in the nervous tissue of L. acuminata. There was, however, no significant change in the level of 5-hydroxyindole acetic acid (5HIAA). The changes were found to be dependent on concentration of the latex extract. Similar changes were produced by both water and organic solvent extracts of the latex. The latex of E. royleana thus affects all the known neurotransmission mechanisms in the snail either separately or through a complex interaction between the different neurotransmitters. This may account for its high toxicity to snails.

Inhibition kinetics of certain organophosphorus and carbamate pesticides on acetylcholinesterase from the snail Lymnaea acuminata.

k2, Kd and ki, for 2 organophosphorus (Phorate and Formothion) and 2 carbamate pesticides (Mexacarbate and Carbaryl) using acetylcholinesterase present in homogenates of the nervous tissue of the snail Lymnaea acuminata, were determined. Calculation of zero time velocities demonstrated that even in their P-S form the organophosphate compounds inhibited snail acetylcholinesterase. The kinetic constants of the 2 carbamates have been explained on the basis of their structure. The toxicity of the 4 pesticides has been explained on the basis of their kinetic constants.

Changes in carbohydrate metabolism in various organs of the snail, Lymnaea acuminata following exposure to trichlorfon.

Several aspects of carbohydrate metabolism following 24 hr and 48 hr treatment with 10 and 20 mg/l of trichlorfon, were studied in hepatopancreas, mantle, intestine and foot of the snail. Lymnaea acuminata. Following treatment with the pesticide, the rate of oxygen consumption and concentration of glycogen were reduced, while the levels of lactic acid and reducing sugars were enhanced. Withdrawal of pesticide for 7 days following trichlorfon treatment (10 mg/l for 48 hrs) could not reverse these changes.

Cyclophosphamide as a potential chemosterilant for harmful snails.

Cyclophosphamide induced changes in the ovotestis of the snail Lymnaea acuminata, the vector of the giant liverflukes Fasciola hepatica and Fasciola gigantica were studied in order to explore potential of the drug as a chemosterilant for snails. The drug caused a dose dependent reduction in the levels of DNA, RNA and proteins and the activity of the enzyme alkaline phosphatase. It increased the activity of acid phosphatase and the levels of total free amino acids in the ovotestis. While the animals showed nearly total recovery in RNA and DNA levels 7 days after termination of drug treatment, changes produced in protein, amino acid levels and phosphatase activity did not show any recovery. It appears that cyclophosphamide, while affecting its primary targets i.e. DNA and RNA, irreversibly inhibits protein synthesis through other cellular enzymes as well.

Chemosterilization and its reversal in the snail Lymnaea acuminata.

Treatment of the snail Lymnaea acuminata, the vector for the liver flukes Fasciola hepatica and Fasciola gigantica, with two alkylating agents (cyclophosphamide and busulfan) and one thiocarbamide (thiourea) caused reduction in the number of eggs and production of non-viable embryos. All the three drugs caused reduction in the levels of DNA, RNA and protein and the activity of alkaline phosphatase in the gonadal tissues of this snail. There was enhancement in free amino acid levels and activity of acid phosphatase by these drugs. Discontinuation of treatment could not reverse the cytotoxic effects of these drugs. Treatment of sterilized snails with prostaglandin E1 (PGE1), prostaglandin F2 alpha (PGF2 alpha) or hydrogen peroxide not only restored the number of eggs but also caused full embryonic development. Also, there was near total recovery of RNA, DNA and protein levels and small changes in free amino acid levels as well as the activity of alkaline and acid phosphatase. It has been suggested that the prostaglandins play a modulatory role in protein synthesis. The possible therapeutic use of PGE1, PGF2 alpha and H2O2 has been pointed out.

Excretion, accumulation and site of synthesis of urea in the snail Pila globosa during active and dormant periods.

The pattern of urea excretion in active and dormant phases of the amphibious Indian apple snail, Pila globosa, as well as the possible site and mechanism of urea synthesis in this animal have been studied. 1. Urea may be synthesized in the hepatopancreas of this snail and excreted through the nephridia with the nephridial fluid during the active period of the animals life. In dormant snails, this metabolic excretory waste accumulates in its various organs and body fluids. 2. The possibility of the presence of an ornithine cycle for the synthesis of urea in the hepatopancreas is indicated in this snail.

Pharmacological studies on the molluscicidal activity of phorate on Pila globosa (Gastropoda).

Amongst heart, rectum and radula protractor muscles of Pila globosa, the heart showed a negative inotropic and negative chronotropic response while the rectum and radula protractor showed a positive tonotropic response to exogenously applied acetycholine (ACh). The anti-cholinesterase pesticide phorate substantially increased the response to ACh; at high doses it could also evoke cholinomimetic response from the heart. The anti-cholinesterase property of phorate gradually increased with an increase in its contact period to the tissue. The extent of alteration in ACh response caused by prior treatment with low and high doses of phorate were very similar, indicating that the efficacy of phorate is by and large time-dependent rather than dose-dependent. The effect of phorate remained irreversible even after prolonged washing. The mode of action of phorate has been discussed in the light of the above findings.

Changes in brain catecholamines and spontaneous locomotor activity in response to thyrotropin releasing hormone.

Repeated exposure of rats to thyrotropin releasing hormone produced a dose- and time-dependent increase in spontaneous locomotor activity accompanied by an increase in brain stem tyrosine hydroxylase. Dopamine levels in cerebral cortex were increased maximally by 34% in animals receiving thyrotropin releasing hormone at a dosage of 2 mg/kg for 10 days. The concentrations of brain stem tyrosine and cerebral cortex norepinephrine remained unaltered in response to thyrotropin releasing hormone treatment. Our data suggest that administration of thyrotropin releasing hormone increases the synthesis and perhaps the turnover of brain catecholamines and that this may constitute an underlying mechanism for the anti-depressant action of this synthetic hormone.

Comparative study of cholinesterase in two snails Pila globosa and Lymnaea acuminata.

Enzymatic activity of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) was measured in the nervous and cardiac tissue of two snails Pila globosa and Lymnaea acuminata. Data on substrate specificity, enzyme kinetic constants and specific enzyme inhibitors indicated that, L. acuminata possessed both AChE and BuChE, while, P. globosa had only AChE. Pharmacological experiments carried out on isolated organs of the two snails indicated that isolated organs of P. globosa were sensitive to low doses of acetylcholine (ACh) and to relatively higher doses of butyrylcholine (BuCh). The responses elicited by ACh could be enhanced by eserine but that of BuCh could not be increased by ethopropazine which has been shown to be a specific inhibitor of BuChE in snails. Isolated hearts of L. acuminata on the other hand, were more sensitive to BuCh as compared to ACh. The BuCh effect was enhanced by ethopropazine while the effect of both ACh and BuCh increased by prior application of eserine. There is a correlation between the occurrence of the two types of cholinesterase (ChE) and the sensitivity of the isolated organs to a specific substrate of these enzymes suggesting a possible correlation between transmitters and cholinesterase system.

Effect of testosterone and 6-hydroxydopamine treatment on the metabolism of catecholamine and 5-hydroxytryptamine in methylcholanthrene-induced prostate carcinoma of rats.

The precursors tyrosine and tryptophan as well as the synthesizing and deaminating enzymes of catecholamines have been identified in methylcholanthrene-induced prostatic carcinoma of rats. Tyrosine hydroxylase, monoamine oxidase, catechol O-methyltransferase, dopamine, 5-hydroxytryptamine, and 5-hydroxyindoleacetic acid seemed to be neoplastic in origin, since electron microscopic studies failed to reveal the presence of any neuronal elements in this squamous epithelial cell carcinoma. Castration of rats significantly reduced the activity of tyrosine hydroxylase and the levels of tyrosine, dopamine, tryptophan, 5-hydroxytryptamine, and 5-hydroxyindoleacetic acid in prostate tumors. The changes appeared to be androgen specific since reintroduction of testosterone restored several of these biochemical parameters virtually to control limits. Chemical sympathectomy induced by 6-hydroxydopamine failed to alter monoamine metabolism; however, the prostatic tumor grown in 6-hydroxydopamine-treated rats showed significantly (32%) less necrosis than those grown in normal animals.