Strain differences in kinetic and thermal stability of two mouse brain tryptophan hydroxylase activities

Levels of forebrain serotonin (5-HT), tryptophan, 5-hydroxyindoleacetic acid (5-HIAA) and hydroxylase cofactor (BH₄) were comparable in two experimental mouse strains (A/J and C57B1/6J) despite 2–3-fold differences in vitro in the relative activities of forebrain and midbrain tryptophan-5-monooxygenase (TPOH; EC 1.14.16.4). The enzyme activities did not differ with respect toK_m for cofactor at saturating levels, but manifested different degrees of cooperativity with respect to cofactor when examined with BH₄ concentrations within a physiological range. They differed also in the frequency and amplitude of kinetic variation around comparable mean velocity slopes across cofactor and time; in resistance to pre-incubation inactivation and responsiveness to its facilitation by calcium; in molecular weight heterogeneity as reflected in the distribution of molecular weight forms by gel diffusion chromatography; and in the number of peaks in power spectral analysis of kinetic variation patterns. Although the potential roles of small-molecule ligand and/or regulator proteins have not been ruled out, we hypothesize that differences in conformational stability underlie the differences in regulatory properties and make one enzyme activity more vulnerable to occlusive influences in vivo.     

Long-term local and distal increases in tryptophan hydroxylase activity following intracerebral kainic acid injections in the rat

The administration of kainic acid (1–2 μg) into the right striatum of adult rats resulted in a marked local increase in tryptophan hydroxylase activity (+ 54–106%). This change was significant as soon as on the second day after the treatment and persisted for at least 12 days. In addition, long-lasting elevations of tryptophan hydroxylase activity were also observed in the anterior raphe area, septum and ipsilateral hippocampus and cerebral cortex. In contrast, the intrahippocampal injection of kainic acid (1 μg) induced a long-term increase in tryptophan hydroxylase activity only in the injected structure. In all cases, the changes in tryptophan hydroxylase activity were associated with significant increases in the V_max of the enzyme with no alteration of its apparent affinities for tryptophan and the pterin cofactor. Studies of the sensitivity of tryptophan hydroxylase from control and from kainic acid-treated rats to in vitro activating conditions (Ca⁺-dependent phosphorylation, partial trypsinization, exposure to sodium dodecyl sulfate) suggest that the intrastriatal injection of the neurotoxin induced a long-lasting activation of the enzyme.These findings indicate that intracerebral injections of kainic acid may be a valuable approach to explore further the mechanisms controlling tryptophan hydroxylase activity in vivo.     

Possibility of using guinea pigs for studying the carcinogenic activity of some endogenous substances

The tryptophan metabolites 3-indolylacrylic acid and 2-aminoacetophenone, whose carcinogenic activity has been demonstrated by experiments on inbred mice, were injected into noninbred guinea pigs. Both substances induced tumors in the animals of the exprimental groups earlier than in the control and the tumors differed significantly in their morphology from those in animals of the control groups, evidence that both compounds have a carcinogenic effect. The results indicate that guinea pigs can be used to study the carcinogenic activity of weak carcinogens of the endogenous class.Experimental Animals Division, Oncologic Scientific Center, Academy of medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 8, pp. 200–202, August, 1977.     

Effects of amitriptyline on serum glutamate and free tryptophan in rats

Summary In rats, chronic amitriptyline (14 days, 10 mg/kg, IP) administration resulted in a significant increase in the serum glutamate concentration and concomitant increase in the serum free tryptophan. In contrast, amitriptyline had no effect on the total serum tryptophan or CSF glutamate level. The data confirmed that antidepressant drugs may induce an increase of the serum glutamate concentration in depressive patients.     

Induction of tyrosine aminotransferase by carcinogenic metabolites of tryptophan and tyrosine

The effect of endogenous carcinogenic substances (3-hydroxyanthranilic and parahydroxyphenyllactic acids) and their noncarcinogenic analogs (anthranilic and phenyllactic acids) on tyrosine aminotransferase activity was compared in rat liver. The carcinogenic metabolites were found to have the property of sharply inducing activity of the enzyme. This phenomenon and existing data on the role of the increase in tyrosine aminotransferase and tryptophan oxygenase activity in tyrosine and tryptophan catabolism in the direction of the possible formation of carcinogenic metabolites suggest that there is a chain reaction of accumulation of endogenous carcinogens in the body.Oncological Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksporimental'noi Biologii i Meditsiny, Vol. 82, No. 9, pp. 1064–1066, September, 1976     

Response to tryptophan depletion in major depression treated with either cognitive therapy or selective serotonin reuptake inhibitor antidepressants

Background

Mood sensitivity to rapid tryptophan depletion (RTD) has been demonstrated in patients treated with antidepressants that act preferentially on the serotonergic system. Depressed patients treated with bright-light therapy also show sensitivity to RTD, but those treated with electroconvulsive therapy or total sleep deprivation do not. Patients treated with an empirically supported psychotherapy have not been investigated for sensitivity to tryptophan depletion. This study compares the effects of RTD in patients treated with either selective serotonin re-uptake inhibitors (SSRIs) or cognitive therapy (CT).

Methods

Twenty patients treated with either SSRIs or CT underwent both rapid tryptophan depletion and sham-depletion using a blinded crossover design. Depressive symptoms were assessed using a modified Hamilton Depression Rating Scale (HDRS) and the Beck Depression Inventory (BDI). The differential change in depression scores across procedures between the groups was compared, and effect sizes were calculated.

Results

The differential worsening of mood for the SSRI group compared with the CT group was significant on the BDI. The effect size of the differential change was 1.6 for the BDI and .8 for the HDRS. Furthermore, the SSRI group experienced significant mood worsening during depletion compared with sham on both the HDRS and the BDI, whereas the CT group did not.

Conclusions

The CT group was resistant to the effects of tryptophan depletion, but the SSRI group was not.     

Transgene expression from the Tribolium castaneum Polyubiquitin promoter

The highly conserved Ubiquitin proteins are expressed from genes with strong, constitutively active promoters in many species, making these promoters attractive candidates for use in driving transgene expression. Here we report the cloning and characterization of the Tribolium castaneum Polyubiquitin (TcPUb) gene. We placed the TcPUb promoter upstream of the coding region of the T. castaneum eye-colour gene Tc vermilion (Tcv) and injected this construct into embryos from a Tcv-deficient strain. Transient expression of Tcv during embryogenesis resulted in complete rescue of the larval mutant phenotype. We then incorporated the TcPUb-Tcv chimera into a piggyBac donor. Resulting germline transformants were easily recognized by rescue of eye pigmentation, illustrating the potential of the TcPUb promoter for use in driving transgene expression.     

Tryptophan depletion effects on EEG and MEG responses suggest serotonergic modulation of auditory involuntary attention in humans

Involuntary attention shifting, i.e., detecting and orienting to unexpected stimulus changes, may be altered at low brain serotonin (5-hydroxytryptamine; 5-HT) levels. This was studied in 13 healthy subjects (21–30 years old; 6 females) by using a dietary challenge, acute tryptophan depletion (ATD), which decreases 5-HT synthesis in the brain. Five hours after ingestion of either ATD or control mixture (randomized, double-blinded, crossover design), brain responses indexing involuntary attention were measured with simultaneous 64-channel electroencephalography (EEG) and 122-channel magnetoencephalography (MEG). During the measurement, the subjects were instructed to discriminate equiprobable 200- and 400-ms tones by pressing one of two buttons rapidly. Occasionally, the frequency of the tones changed (10% increase/decrease), causing involuntary attention shifting. ATD significantly lowered plasma tryptophan concentrations (total tryptophan decreased by 75%, free tryptophan decreased by 35%). As compared to the control condition, ATD reduced the amplitude of the deviant-tone N2 wave, including the overlapping mismatch negativity (MMN) and N2b subcomponents, which are suggested to reflect change detection in the brain. The EEG results were accompanied by a significant increase in the peak latency of the magnetic counterpart of MMN. However, no ATD effects were observed in P3 to task-irrelevant frequency change. Reaction time (RT) to deviants per se was not significantly affected, but RT in trials succeeding the deviant-frequency tones was increased by ATD, which suggested impaired reorienting to the task-relevant activity. In conclusion, the results suggest that decreased level of central 5-HT function after ATD may decrease involuntary attention shifting to task-irrelevant sound changes and thus modulate resource allocation to the task-relevant activity.     

Eating ourselves to death (and despair): The contribution of adiposity and inflammation to depression

Obesity and related metabolic conditions are of epidemic proportions in most of the world, affecting both adults and children. The accumulation of lipids in the body in the form of white adipose tissue in the abdomen is now known to activate innate immune mechanisms. Lipid accumulation causes adipocytes to directly secrete the cytokines interleukin (IL) 6 and tumor necrosis factor α (TNFα), but also monocyte chemoattractant protein 1 (MCP-1), which results in the accumulation of leukocytes in fat tissue. This sets up a chronic inflammatory state which is known to mediate the association between obesity and conditions such as cardiovascular disease, type 2 diabetes, and cancer. There is also a substantial literature linking inflammation with risk for depression. This includes the observations that: (1) people with inflammatory diseases such as multiple sclerosis, cardiovascular disease, and psoriasis have elevated rates of depression; (2) many people administered inflammatory cytokines such as interferon α develop depression that is indistinguishable from depression in non-medically ill populations; (3) a significant proportion of depressed persons show upregulation of inflammatory factors such as IL-6, C-reactive protein, and TNFα; (4) inflammatory cytokines can interact with virtually every pathophysiologic domain relevant to depression, including neurotransmitter metabolism, neuroendocrine function, and synaptic plasticity. While many factors may contribute to the association between inflammatory mediators and depression, we hypothesize that increased adiposity may be one causal pathway. Mediational analysis suggests a bi-directional association between adiposity and depression, with inflammation possibly playing an intermediary role.     

Plasma Ratios of Tryptophan and Tyrosine to Other Large Neutral Amino Acids in Manic-Depressive Patients

Abstract: The plasma ratio of each neutral amino acid (tryptophan (TRP), tyrosine (TYR), valine, isoleucine, leucine (LEU) or phenylalanine) to the sum of the other neutral amino acids was measured in 16 : manic and 14 : depressed patients. In the manics, there was a correlation between the psychomotor activity and the plasma TRP and LEU ratios. In the depressives, the depressed mood, retardation and global severity were correlated with the TRP ratio. The zotepine responders showed an increase in the TRP ratio after treatment. In the mianserin responders, the TYR ratio, which was high before the treatment, decreased to the normal range after the treatment. But, the plasma amino acid ratios remained unchanged in the patients treated with lithium carbonate or amitriptyhe. These results suggest that, in manic-depressive illness, there might be abnormalities in the metabolism of neutral amino acids, mainly of TRP and TYR, and that the plasma TRP and TYR ratios might be important indicators for determining the efflcacy of some drugs.