Kensuke Kameda

Serotonin 5-HT2A receptor gene polymorphism, 5-HT2A receptor function and personality traits in healthy subjects: a negative study

BACKGROUND Central serotonin-2A (5-HT(2A)) receptor dysfunction is regarded as an important factor in the etiology of affective disorders. The relations between some personality traits and the vulnerability of affective disorders are also implicated. Moreover, there are several reports which describe the association between 5-HT(2A) receptor gene polymorphisms and mental disorders. We therefore examined the relationship between personality traits, the 5-HT(2A) receptor function, and 5-HT(2A) receptor gene polymorphisms. METHODS 5-HT-induced intraplatelet calcium (Ca) mobilization, 5-HT(2A) receptor gene polymorphisms (A-1438G, T102C, T516C, C1340T, C1354T), and Temperament and Character Inventory (TCI) scores were examined in 133 healthy subjects. RESULTS Neither 5-HT-induced Ca mobilization nor 5-HT(2A) receptor gene polymorphisms (A-1438G, T102C) appear to be associated with seven personality dimensions including Harm Avoidance. There was no significant difference in the Ca response among the subjects with -1438A/A, A/G and G/G genotypes. Since the appearance of the other types of the 5-HT(2A) receptor gene polymorphisms (T516C, C1340T and C1354T) was quite rare in our sample, we were unable to examine the relationship between these polymorphisms, and the TCI score or the Ca response. LIMITATIONS Our failure to find a significant association may reflect the false negative results due to the small sample size and low statistical power. Further studies in depressed patients may clarify the complicated relationship between personality traits and the vulnerability of affective disorders. CONCLUSIONS Personality traits detected by TCI may not be directly related to the 5-HT(2A) receptor function or 5-HT(2A) receptor gene polymorphism which may be involved in the vulnerability of affective disorders.

Differential effects of subchronic treatments with atypical antipsychotic drugs on dopamine D2 and serotonin 5-HT2A receptors in the rat brain.

Summary. The effects of 3-week treatment with a typical antipsychotic drug chlorpromazine and three atypical antipsychotic drugs (risperidone, olanzapine and perospirone) on the binding to dopamine D2 and serotonin 5-HT2A receptors were examined in the rat stratum and frontal cortex, respectively. Subchronic treatment with chlorpromazine (10 mg/kg) and perospirone (1 mg/kg) significantly increased D2 receptors, while no increase was observed with lower dose of chlorpromazine (5 mg/kg), perospirone (0.1 mg/kg), risperidone (0.25, 0.5 mg/kg) or olanzapine (1, 2 mg/kg). On the other hand, 3-week administration of chlorpromazine (5, 10 mg/kg) and olanzapine (1, 2 mg/kg) significantly decreased 5-HT2A receptors, but risperidone (0.25, 0.5 mg/kg) or perospirone (0.1, 1 mg/kg) had no effect. The measurement of in vivo drug occupation for D2 and 5-HT2A receptors using N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) suggested that high occupation of 5-HT2A receptors with lower D2 receptor occupancy might be involved in the absence of up-regulation of D2 receptors after subchronic treatment with some atypical antipsychotic drugs.

Effects of lithium on dopamine D2 receptor expression in the rat brain striatum

Summary. Effects of lithium on the dopamine D2 receptor expression in the rat brain striatum were studied. Feeding the chow containing 0.2% LiCO3 for 6 days increased the level of the dopamine D2 receptor mRNA, and the transcription rate of the dopamine D2 receptor gene, indicating the stimulatory effects of lithium on the transcription of the dopamine D2 receptor gene. [3H] Spiperone binding to the striatal membranes increased in the rats treated with lithium, while the Western blotting analysis showed no change of the amount of the dopamine D2 receptors. These results suggested that lithium might induce the conformational changes of the dopamine D2 receptors. The methamphetamine-induced locomotor activity was enhanced by the pretreatment with lithium, whereas simultaneous increase in the methamphetamine concentration in the striatum was also observed. These observations suggested that the stimulation of methamphetamine-induced locomotor activi-ty by lithium might be, at least partly, due to either increased sensitivity of the dopamine receptors, or increased concentration of methamphetamine in brain, or combination of both.

Further purification, characterization and salt activation of acyl-CoA synthetase fromEscherichia coli

Acyl-CoA synthetase was further purified from Escherichia coli in good yield and fold purification by affinity chromatography on CoA-Sepharose 4B. The molecular weight of the active form of the purified enzyme was estimated as 45 000 by Sephadex G-100 and 47 000 by Sephadex G-200. Sedimentation equilibrium ultracentrifugation analysis revealed a molecular weight of 50 000. The sedimentation coefficient was calculated as 4.4 S. An absorption maximum at 276 nm was observed in the ultraviolet light absorption spectrum. The molar extinction coefficient was 9.2 X 10(4). Kinetic constants were determined for trans fatty acids. All ions tested, including chaotropic and lyotropic ions, stimulated or inhibited acyl-CoA synthetase activity depending on their concentrations in the assay system. In a series of chaotropes, the lower concentration required to maximally activate acyl-CoA synthetase in increasing order of potency of chaotropic ions. The inhibitory effect of chaotrope on the enzyme activity was reversible. These data suggest that salts have a common mode of action and influence acyl-CoA synthetase activity primarily through their effect on the solution structure.

The activity of fatty acid synthase of epidermal keratinocytes is regulated in the lower stratum spinousum and the stratum basale by local inflammation rather than by circulating hormones

The epidermal keratinocytes produce and secrete lipids to maintain the water barrier of the epidermis. To clarify the regulation of epidermal lipid synthesis, we investigated the hormonal effect on the activity of fatty acid synthase (FAS) of the keratinocytes, and the expression of FAS in the human skin. In cultured keratinocytes, the FAS activity, assayed by measuring the oxidation of NADPH, was slightly increased by hydrocortisone or testosterone, but not influenced by thyroid hormone, estrogen, progesterone or insulin. In immunohistochemical study of normal human epidermis, FAS was expressed strongly in the stratum granulosum and moderately in the uppermost layer of the stratum spinousum (SS), suggesting that fatty acid synthesis may increase during normal epidermal differentiation. In inflammatory disorders, such as psoriasis, lichen planus, and atopic dermatitis, FAS was also expressed in the lower SS and the stratum basale (SB), resulting in strong staining in the whole layers of the epidermis. Remarkable increase of FAS expression was only observed in the lower SS and the SB. Therefore, the activity of FAS in the epidermis may be regulated in the lower SS and the SB by local inflammation rather than by circulating hormones. In other components of the skin, FAS was strongly expressed not only in adipose tissue and sebaceous glands, which are known as active sites of lipid synthesis, but also in sweat glands, suggesting that the sweat glands can synthesize abundant fatty acids de novo.

Treatment response in depressed patients with enhanced Ca mobilization stimulated by serotonin

Serotonin (5-HT)-stimulated intraplatelet calcium (Ca) mobilization has been shown to be enhanced in nonmedicated depressive patients by many studies. However, there has not been any longitudinal follow-up study of this parameter. We examined the relationship between treatment response and pretreatment value of the 5-HT-induced Ca response. The 5-HT(10 μM)-induced intraplatelet Ca mobilization was measured in 98 nonmedicated depressive patients (24 bipolar disorders, 51 melancholic major depressive disorders, and 23 non-melancholic major depressive disorders). These patients were followed up prospectively for a further period of five years. The depressed patients with enhanced Ca response to 5-HT in bipolar disorders exhibited a good response to mood stabilizers but those with major depressive disorders showed a poor response to antidepressants. These findings suggest the possibility that the 5-HT-induced intraplatelet Ca response may be a good predictor of treatment response in depressed patients. Longer longitudinal follow-up studies are needed in larger samples to examine if this parameter may be a specific biological marker for unipolar-bipolar dichotomy.

Effects of citalopram on dopamine D2 receptor expression in the rat brain striatum.

Effects of citalopram on dopamine D2 receptor expression in the rat brain striatum were studied. Repeated administration of citalopram increased the amount of dopamine D2 receptors, the level of dopamine D2 receptor mRNA, and the transcription rate of the dopamine D2 receptor gene. Single administration of citalopram also increased the level of dopamine D2 receptor mRNA with a maximum effect in 2–4 h after the treatment, and the transcription rate of the dopamine D2 receptor gene. The administration of 5-hydroxytryptophan (5-HTP) also increased the level of dopamine D2 receptor mRNA. These results suggest that the increase in the dopamine D2 receptor expression induced by citalopram may be owing, at least partially, to the stimulation of the dopamine D2 receptor gene transcription, and that serotonin (5-HT) may mediate the effects of citalopram in the induction of dopamine D2 receptor expression.

Isolation and characterization of the multiple charge isoforms of acyl-CoA synthetase from Escherichia coli

The three fractions of acyl-CoA synthetase differing in isoelectric pH were isolated from Escherichia coli by isoelectric focusing and characterized. They had the same molecular weight and identical immunochemical properties. The three fractions differed appreciably in pH-velocity profiles. These fractions had distinguishable thermal stabilities and peptide patterns obtained after limited proteolysis. Apparent Km and Vmax values for fatty acids were also significantly different in these fractions, although the specificity ranged from C8 to C18 fatty acids with maximum activity for lauric acid in all fractions.

Effects of acute citalopram treatment on the methamphetamine-induced locomotor activity

1. Previously the authors have shown that acute citalopram treatment increased the dopamine D2 receptor expression in rat brain striatum (Kameda et al., 2000). In the present study, the authors attempted to determine whether these effects of citalopram influence the methamphetamine-induced locomotor activity. 2. The pretreatment with a single administration of citalopram (10 mg/kg, i.p.) resulted in the significant enhancement of the locomoter activity induced by methamphetamine treatment (1 mg/kg, i.p.). The enhancement was observed 30 min, 12 hours, 24 hours, but not 7 days after withdrawal of citalopram administration. 3. Then the authors determined the methamphetamine concentration in rat brain striatum by gas chromatography-mass spectrometry (GC-MS) The results showed that the concentration of methamphetamine wars significantly higher in the rats 24 hours, and also 7 days after withdrawal of citalopram administration, compared to the control rats. 4. These results emphasized the involvement of the high methamphetamine concentration, caused by the pretreatment with citalopram, in the enhancement of the methamphetamine-induced locomotor activity. However high methamphetamine concentration alone could not account for this enhancement, since the high concentration of methamphetamine observed 7 days after withdrawal of citalopram administration did not appear to enhance the methamphetamine-induced locomotor activity. Another mechanism through which the pretreatment with citalopram enhanced the methamphetamine-induced locomotor activity, such as the increased expression of the dopamine D2 receptors, could not be excluded.