Quantification of α1-Adrenoceptor Subtypes in Human Tissues by Competitive RT-PCR Analysis

Faure, Céline; Gouhier, C.; Langer, S.Z.; Graham, Dawn

doi:10.1006/bbrc.1995.2219

Cited by 42 publications

(23 citation statements)

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“…Using a polymerase chain reaction-MIMIC construction kit (Clontech, Palo Alto, CA, USA) an internal DNA standard was constructed (Engstrøm et al 1997). The internal standard RNA was constructed mainly as described by Faure et al (1995). A composite primer, comprising 37 nucleotides of bacteriophage T7 RNA-polymerase promoter followed by the sequences of our usual sense primer, was used for amplification of the DNA sequence by PCR.…”

Section: Primers and Construction Of Internal Mrna Standardmentioning

confidence: 99%

Desensitization of beta2-adrenoceptor function in non-pregnant rat myometrium is modulated by sex steroids

Engstrøm

Vilhardt²,

Bratholm³

et al. 2001

Journal of Endocrinology

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The effects of in vivo treatment with estrogen and progesterone on isoproterenol-induced uterine relaxation and 2 -adrenoceptor ( 2 AR) mRNA production in nonpregnant rat myometrium were investigated. Whether homologous myometrial desensitization of 2 AR function was dependent on or modulated by the two steroids was also examined.Estrogen treatment alone or in combination with progesterone reduced maximal relaxation (E max ) of isolated uterine strips subsequently challenged with isoproterenol whereas progesterone alone had no effect on this parameter. The reduction was accompanied by an enhanced 2 AR mRNA concentration. The concentration of isoproterenol giving half-maximal relaxing response (EC 50 ) increased following estrogen treatment and this effect was curbed by progesterone.Isoproterenol had no effect on 2 AR transcription irrespective of the steroid regimes employed. E max of isolated uterine strips was reduced following prolonged in vivo treatment with isoproterenol but the effect was found only when estrogen alone was administered concomitantly. Finally, in vivo treatment with isoproterenol increased EC 50 of uterine strips subsequently stimulated with isoproterenol in vitro. This effect was independent of steroid treatment.We conclude that homologous desensitization of 2 AR function in non-pregnant rat myometrium in terms of sensitivity (EC 50 ) is independent of sex steroids but in terms of maximal response (E max ) occurs only in the presence of estrogen. We speculate whether progesterone withdrawal in connection with the well-known estrogen dominance at rat parturition may strengthen the desensitization induced by 2 AR activation and thus contribute to the transformation of the uterus from a quiescent to a highly contractile organ.

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Section: Primers and Construction Of Internal Mrna Standardmentioning

confidence: 99%

Desensitization of beta2-adrenoceptor function in non-pregnant rat myometrium is modulated by sex steroids

Engstrøm

Vilhardt²,

Bratholm³

et al. 2001

Journal of Endocrinology

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“…An internal standard RNA for the GCR mRNA, the b 2 -adrenoceptor mRNA and the c-fos mRNA was constructed mainly as described by Faure et al 6 The resulting internal standard RNA was quanti®ed by UV-detection (Gene-Quant II, Pharmacia, Sweden). The resulting RT-PCR products were indistinguishable from the internal standard DNA.…”

Section: Subjects and Protocolmentioning

confidence: 99%

Lymphocyte glucocorticoid receptor mRNA correlates negatively to serum leptin in normal weight subjects

et al. 2000

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OBJECTIVE: The aim of the present study was to test the hypothesis that glucocorticoid receptor mRNA concentrations decreased with increasing fatness in normal subjects. MEASUREMENTS: Serum leptin concentrations, fat distribution parameters, lymphocyte glucocorticoid (GCR) mRNA, b b 2 -adrenoceptor mRNA and c-fos mRNA concentrations measured by RT-PCR-HPLC. SUBJECTS: Fifteen healthy non-obese young subjects with a mean body mass index (BMI) of 23.5 AE 0.3 ( AE s.e.m.) kgam 2 . RESULTS: Lymphocyte GCR and b b 2 -adrenoceptor mRNA concentrations averaged 4.2 AE 0.2 ( AE s.e.m.) amolam mg total RNA and 1.4 AE 0.1 amolam mg total RNA, respectively. There was a signi®cant negative correlation between serum leptin and lymphocyte GCR mRNA (P`0.01). Serum leptin correlated positively with the waist ± hip ratio (P`0.03), whereas lymphocyte GCR mRNA correlated negatively to the waist ± hip ratio (P`0.04). Serum cortisol correlated with the weight of the subjects but not the waist ± hip ratio or GCR mRNA. CONCLUSIONS: It is suggested that the decrease in lymphocyte GCR mRNA concentration with increasing serum leptin concentrations is a counterregulatory response to an increased body fat content. Further studies are warranted, especially to elucidate the relationship between GCR mRNA in lymphocytes and in fat cells and to clarify the mechanism of the decrease in GCR mRNA.

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“…The internal standard RNA was constructed mainly as described by Faure et al 10 The resulting internal standard RNA was quanti®ed by UV-detection (Gene-Quant, Pharmacia, Sweden). The resulting RT-PCR product was indistinguishable from the internal standard DNA.…”

Section: Flowcytometric Analysis Of Subset Frequenciesmentioning

confidence: 99%

Obese male subjects show increased resting forearm venous plasma noradrenaline concentration but decreased 24-hour sympathetic activity as evaluated by thrombocyte noradrenaline measurements

Verdich²,

Astrup³

et al. 1999

Int J Obes

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OBJECTIVE: To study resting forearm venous plasma noradrenaline (NA) and 24 h sympathoadrenal activity evaluated by measurements of thrombocyte NA and adrenaline (A) in obese male subjects before and after weight reduction. DESIGN: Blood samples were collected in obese subjects and in controls after an overnight fast. MEASUREMENTS: Fatness and fat distribution parameters, plasma and thrombocyte NA and A, lymphocyte b b 2 -adrenoceptor mRNA, cAMP and lymphocyte subset composition. RESULTS: Forearm venous plasma NA at rest was signi®cantly elevated in the obese subjects and correlated to the body mass index and diastolic blood pressure. Thrombocyte NA and A correlated, however, negatively to the body fat % in obese subjects. Further analysis showed that thrombocyte NA and A were reduced in obese subjects with body fat % above 40% but not in the other groups. Weight reduction normalized both forearm venous plasma NA and thrombocyte NA and A. b b 2 -adrenoceptor mRNA correlated positively to the frequency of CD3 7 CD56 and CD3 CD8 cells in the blood and negatively to plasma A, but there was no difference in the mRNA level, cyclic AMP and lymphocyte subset composition between obese and controls. CONCLUSIONS: Sympathetic activity at rest as evaluated by forearm venous plasma NA was increased in the obese male subjects and this abnormality was related to the increase in body mass index and the arterial blood pressure. Thrombocyte NA and A, which is likely to re¯ect 24 h plasma catecholamine concentrations were reduced in obese subjects with a body fat % above 40%, probably due to a reduced physical activity in these subjects. Both catecholamine parameters were normalized by weight reduction.

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Quantification of α1-Adrenoceptor Subtypes in Human Tissues by Competitive RT-PCR Analysis

Cited by 42 publications

References 0 publications

Desensitization of beta2-adrenoceptor function in non-pregnant rat myometrium is modulated by sex steroids

Desensitization of beta2-adrenoceptor function in non-pregnant rat myometrium is modulated by sex steroids

Lymphocyte glucocorticoid receptor mRNA correlates negatively to serum leptin in normal weight subjects

Obese male subjects show increased resting forearm venous plasma noradrenaline concentration but decreased 24-hour sympathetic activity as evaluated by thrombocyte noradrenaline measurements

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