Beta 3-adrenoceptors: their role and regulation in the gastrointestinal tract.
Explore the source record for details and available documents.
Adipose Tissue↗
Biomedical subjects
B A Evans
Publications and source records attributed to B A Evans.
Mapping the gene for sex-linked anemia: an inherited defect of intestinal iron absorption in the mouse.
The sex-linked anemic (sla) mouse carries an anemia that results from an inherited defect of intestinal iron absorption and provides an ideal model with which to investigate this poorly understood yet clinically important process. We have precisely mapped the sla locus within the central region of the X chromosome in relation to a panel of microsatellite markers. Analysis of over 500 progeny from an intraspecific intercross and a smaller intraspecific backcross segregating sla established the following locus order in the sla region: DXMit45-sla- (DXMit16, DXMit96)-DXMit41-DXMit169-DXMit170- DXMit148-(DXMit18, DXMit171)-DXMit84-DXMit64. The two microsatellites DXMit16 and DXMit96 are located 0.60 +/- 0.35cM from sla and form the telomeric limit of the sla region. The mapping of the sla locus is an important first step to identifying the gene itself.
Anemia, Iron-Deficiency↗
In vitro patch-clamp studies in skin fibroblasts.
We have conducted single-channel patch-clamp experiments in skin fibroblasts maintained in culture. Two different cell lines, a mouse 3T3-L1 cell line and a human B17 cell line, were selected for these pilot studies. Recordings were made from both cell-attached and excised inside-out patches at room temperature. In the case of the 3T3-L1 cells, the success rate in obtaining good seals (> 1Gomega) was low, and channel openings in either cell-attached or excised patches were rare. We have, however, identified a channel in a cell-attached configuration with a slope conductance of 39 pS in symmetrical K+ solutions. In the case of the human B17 cells, good quality seals were more readily obtained. One principal type of channel opening was identified. In cell-attached patches, the prevalent type of channel in symmetrical K+ solutions had a conductance of 187 pS. This channel was activated by strong depolarization, and there was usually more than one active channel in the patch. It was blocked by extracellular tetraethylammonium (20 mM), and persisted when external Cl- was replaced by aspartate. In excised inside-out patches bathed in symmetrical K+, this channel was activated by an increase in Ca+ applied to the intracellular face. A large conductance channel (175 pS) was also observed in excised inside-out patches, with a reverse physiological K+ gradient. This channel had a reversal potential > 40 mV and appeared not to be voltage-dependent under these recording conditions (2 mM Ca(2+)i). We conclude that the channel we have identified in these cells belongs to the maxi-K+ channel class.
3T3 Cells↗
Differential regulation of beta3-adrenoceptors in gut and adipose tissue of genetically obese (ob/ob) C57BL/6J-mice.
1. Levels of beta3-adrenoceptor (AR) mRNA were compared using reverse transcription-polymerase chain reaction (RT PCR) in white adipose tissue (WAT), brown adipose tissue (BAT), ileum and colon from genetically obese (ob/ob) and lean (+/+) C57BL/6J mice. Functional responses to the beta3-AR agonist CL 316243 were also characterized in ileal longitudinal smooth muscle from obese and lean mice. 2. Beta3-AR mRNA levels were significantly higher in WAT (100+/-16%) and BAT (100+/-13%) from lean compared to WAT (21.0+/-0.9%; n=4; P<0.005) and BAT (14.1+/-2.2%; n=5; P<0.01) from obese mice. In contrast, beta3-mRNA levels were not significantly different in ileum (100+/-15%) and colon (100+/-22%) from lean mice, compared to ileum (78+/-13%; n=4; P= 0.31) or colon (82+/-15%; n =4; P=0.52) from obese mice. 3. Concentration-response curves to CL 316243 did not differ significantly in slope or position in ileal longitudinal smooth muscle from obese or lean mice. pEC50 (+/-s.e.mean) values were not significantly different (P= 0.59) between obese (7.90+/-0.13, n = 7) and lean (7.77+/-0.20, n = 7) mice. 4. pKB values for the beta1-AR and beta2-AR selective antagonist propranolol or the beta3-AR selective antagonist SR 58894 against relaxations to CL 316243 were similar in ileum of genetically obese (propranolol 6.31+/-0.22 and 6.13+/-0.12; SR 58894 8.22+/-0.06) and lean mice (propranolol 6.40+/-0.08 and 6.60+/-0.13; SR 58894 8.27+/-0.12) and were consistent with values previously found at beta3-AR. 5. Treatment of lean C57BL/6J mice with dexamethasone (1 mg kg(-1), i.p.) significantly reduced beta3-AR mRNA levels after 4 h in WAT (100+/-6.1 to 41.4+/-4.3; n= 16 18; P<0.0001) and BAT (100+/-8.0 to 35.1+/-5.8; n= 17; P<0.0001), but caused no change in ileum (100+/-6.1 to 101+/-17; n= 10-11; P=0.95) or colon (100+/-11 to 101+/-11; n= 11; P= 0.94). Beta3-mRNA levels in ileum and colon also did not change significantly when examined over 24 h or after the administration of a higher dose of dexamethasone (5 mg kg(-1)). 6. In summary, beta3-AR mRNA levels were considerably lower in WAT and BAT of obese compared to lean mice whereas the levels in ileum and colon were not significantly different. The similar beta3-mRNA levels in ileum of obese and lean mice were associated with indistinguishable responses of carbachol-contracted ileum to a beta3-agonist and similar affinity for beta-antagonists. Administration of glucocorticoids to lean mice reduced beta3-AR mRNA levels in WAT and BAT but not in ileum or colon. These studies show that in mice, beta3-ARs are differentially regulated in ileum and colon compared to adipose tissues.
Adipose Tissue↗
Racial origin, sexual behaviour, and genital infection among heterosexual men attending a genitourinary medicine clinic in London (1993-4).
OBJECTIVES: To compare variables of sexual behaviour and incidence of genital infections among heterosexual men of different racial origins. DESIGN: A prospective cross sectional study of sexual behaviour reported by a standardised self administered questionnaire in new patients who presented for screening and diagnosis. SETTING: A genitourinary medicine clinic in west London. SUBJECTS: 1212 consecutive heterosexual men newly attending in 1993-4. MAIN OUTCOME MEASURES: Variables relating to sociodemographic status, sexual behaviour, condom use, sexually transmitted diseases, and other genital infections stratified by racial origin. RESULTS: There were 941 evaluable heterosexual men of whom the majority were white (79%) and 17% were black. The black men comprised more teenagers (11% cf 2%; p < 0.00001), were more likely to be unemployed (26% cf 12%; p < 0.00001), to have commenced intercourse much earlier (45% cf 22% before aged 16: p < 0.0001), and to have had intercourse with an African woman (14% cf 6%; p < 0.001). Both fellatio (64% cf 96%; p < 0.00001) and cunnilingus (40% cf 92%; p < 0.00001) were practised less frequently by the black men and so too was anal intercourse (11% cf 27%; p < 0.00001). Similar proportions from both groups were non-smokers (53% cf 57%), but a significantly higher proportion of the black men did not drink alcohol (13% cf 5%; p < 0.001). Gonorrhoea (15% cf 1%; p < 0.00001), chlamydial infection (17% cf 8%; p < 0.001), and non-gonococcal urethritis (37% cf 24%; p = 0.001) were diagnosed more frequently in the black men. These findings remained significant after logistic regression and are therefore independently associated with black race. However, there was no significant difference in numbers of sexual partners in the preceding year (median 2), nor in condom use with regular and non-regular partners. The Asian men had commenced intercourse later (mean 19.1 years) than both the black men (mean 15.9 years) and the white men (mean 17.3 years). CONCLUSIONS: Compared with white men, black men attending a genitourinary medicine clinic were much more likely to be unemployed, to have commenced intercourse earlier and to have urethral infection. They were much less likely to practice fellatio, cunnilingus, or anal intercourse. However, there was no difference between the two racial groups in respect of numbers of sexual partners and condom use.
Adolescent↗
Racial origin, sexual lifestyle, and genital infection among women attending a genitourinary medicine clinic in London (1992).
OBJECTIVES: To compare variables of sexual behaviour and incidence of genital infections among women of different racial origins and lifestyles. DESIGN: A prospective cross sectional study of sexual behaviour reported by a standardised self administered questionnaire in new patients who presented for screening and diagnosis. SETTING: A genitourinary medicine clinic in west London. SUBJECTS: 1084 consecutive women newly attending in 1992. MAIN OUTCOME MEASURES: Variables relating to sociodemographic status, sexual lifestyle, condom use, sexually transmitted diseases, and other genital infections stratified by racial origin. RESULTS: There were 948 evaluable women, of whom 932 (98.3%) were heterosexual and 16 (1.7%) were lesbian. Previous heterosexual intercourse was reported by 69% of lesbian women and their most frequent diagnosis was bacterial vaginosis (38%). The majority of heterosexual women were white (78%) and 16% were black. The black women were more likely to be teenagers (18% cf 8%; p = 0.0004) or students (28% cf 15%; p = 0.0008), and to have had an earlier coitarche (48% cf 38% before aged 17; p < 0.004). They also had a higher proportion of pregnancies (58% cf 38%; p < 0.00001) and births (38% cf 20%; p < 0.00001). The white women showed significantly more sexual partners during the preceding year (p = 0.004) and in total (p < 0.00001) and more reported non-regular partners (48% cf 35%; p = 0.004) with whom they were more likely to use condoms (p = 0.009). However, the black women were more likely to have gonorrhoea (7% cf 2% p < 0.0003), chlamydial infection (12% cf 5% p < 0.002), trichomoniasis (10% cf 2% p < 0.00001), or to sexual contacts of men with non-gonococcal urethritis (19% cf 12% p < 0.02). They were less likely to have genital warts (3% cf 12% p = 0.002). Logistic regression showed that all these variables were independently associated with the black women. The Asian women (2%), none of whom had a sexually transmitted disease, had commenced intercourse later (mean 19.7 years) than both black women (mean 16.8 years) and white women (mean 17.6 years). CONCLUSIONS: Sexual intercourse commenced approximately 1 year earlier in the black women, who were more likely to have become pregnant, had children, and to have acquired a bacterial sexually transmitted infection than were the white women.
Adolescent↗
Disorders of fat distribution in HIV infection.
Alterations in body shape due to fat loss and/or redistribution have been described in HIV-infected individuals and associated with the use of antiretroviral (ARV) combination therapy. Certain of these changes have been referred to as peripheral lipodystrophy (LD) and we describe 12 patients who were recognized with this condition between September 1997 and February 1998. It occurred in 12.5% of patients on ARV combination therapy that included a protease inhibitor (PI). In early descriptions the emphasis was on the abdomen, which may be grossly enlarged. In our patients this feature was much less marked. Patients with LD were significantly older than those on PI therapy who did not develop this condition (P=0.016). Although all had raised triglyceride (TG) levels, the elevations were not severe (maximum=6.3 mmol/l). CD4 lymphocyte and viral load levels suggested an optimal response to ARV therapy at the time LD developed. Appearances may be disfiguring but no serious systemic consequences of LD have been observed. Most individuals have chosen to remain on their present ARV combinations. When LD occurs, it appears to be a marker of effective response to anti-HIV therapy.
Adult↗
Heterosexual behaviour, risk factors and sexually transmitted infections among self-classified homosexual and bisexual men.
Our study of men presenting at a genitourinary medicine clinic shows that self-classification into homosexual or bisexual does not accurately define behaviour. We found that 8.5% of self-defined homosexual men had had heterosexual intercourse in the past year and that 26% of self-defined bisexual men had not. Overall, 19% of homosexual/bisexual men reported vaginal intercourse in the past year and a further 42% in their lifetime. Compared with heterosexual men attending our clinic, the practising bisexual men were significantly more likely to come from a white ethnic group (P < 0.003) and to use condoms invariably with regular female partners (P = 0.0001). There was no significant difference in consent for HIV testing between homosexual (43%), practising bisexual (49%) and heterosexual (42%) men despite significantly different perceptions of risk. None of the practising bisexual men was seropositive for HIV infection (P = 0.06) or for syphilis (P = 0.02), or had chlamydial infection, which was found infrequently among homosexual men in general (P = 0.00001). HIV infection found in 19.4% of the exclusively homosexual men was associated with more frequent alcohol consumption (P=0.06).
Adult↗
Central levodopa metabolism in Parkinson's disease after administration of stable isotope-labeled levodopa.
We report the use of a new stable isotope-labeled form of levodopa (LD) to examine in vivo central LD metabolism in Parkinson's disease (PD). Eight patients representing a wide spectrum of disease severity were administered 50 mg of carbidopa orally followed in 1 hour by an intravenous bolus of 150 mg of stable isotope-labeled LD (ring-1',2',3',4',5',6'-(13)C6). Serial blood samples were taken every 30 to 60 minutes and a lumbar puncture was performed 6 hours after the infusion. The average percentage of labeled homovanillic acid (HVA) in lumbar cerebrospinal fluid (CSF) was 54% (SD, 9%; range, 34-67%). The mean CSF labeled HVA concentration was 34.7 ng/ml (SD, 20.2 ng/ml; range, 11.3-67.9 ng/ml). Area under the curve for labeled serum LD closely predicted CSF labeled HVA concentrations (r = 0.747, p = 0.033). Labeled CSF HVA levels did not significantly correlate with either quality or duration of response to the labeled LD dose. In a similar manner, labeled CSF HVA concentrations were not influenced by duration of disease or previous daily LD dosage. These findings support the hypothesis that levodopa-induced benefit in PD is not severely limited by a defect in central levodopa metabolism.
Adult↗
Functional and molecular evidence for beta 1-, beta 2- and beta 3- adrenoceptors in human colon.
1. Relaxation of carbachol pre-contracted human colonic muscle to (-)-isoprenaline was examined in circular, longitudinal and taenia coli preparations to determine the beta-adrenoceptor subtypes involved. beta 1-, beta 2- and beta 3-Adrenoceptor mRNAs were also measured in colonic muscle and mucosa. 2. (-)-isoprenaline caused relaxation of longitudinal smooth muscle preparations with pEC50-7.39 +/- 0.12, and this response was inhibited by both propranolol (0.1 microM, pKB 8.55 +/- 0.12) and the selective beta 1-antagonist, CGP 20712A (0.1 microM, pKB 8.80 +/- 0.20), while the selective beta 2-antagonist, ICI 118551 (0.1 microM) failed to inhibit isoprenaline relaxation consistently. 3. (-)-Isoprenaline caused relaxation of taenia coli with a pEC50 of 6.70 +/- 0.17. Propranolol (0.1 microM). CGP 20712A (0.1 microM) and ICI 118551 (0.1 microM) inhibited the isoprenaline response with similar low affinities (pKB values 7.93, 7.71 and 7.54, respectively). Carbachol pre-contracted circular smooth muscle preparations failed to relax consistently to isoprenaline and these responses were not characterized. 4. beta 1- and beta 2-Adrenoceptor mRNAs were present in circular longitudinal muscle samples and taenia coli samples, and lower levels were detected in mucosa. beta 3-mRNA was also present in both muscle preparations but was not detected in human colonic mucosa. 5. In summary, beta 1-adrenoceptors are the predominant subtype mediating isoprenaline-induced relaxation of the thin longitudinal smooth muscle of human colon, while beta 1-receptors do not appear to be involved in these responses. However, beta 3-adrenoceptors may play a role in relaxation of the taenia coli as conventional antagonist affinities are low. beta 3-Adrenoceptor mRNA was present in taenia coli and circular/longitudinal smooth muscle but absent from human colonic mucosa.
Adrenergic beta-Agonists↗
Evaluation of sorivudine (BV-araU) versus acyclovir in the treatment of acute localized herpes zoster in human immunodeficiency virus-infected adults. The Multinational Sorivudine Study Group.
The clinical efficacy and safety of sorivudine as treatment for acute cutaneous zoster in human immunodeficiency virus-infected adults was compared with that of acyclovir in a double-blinded randomized study. A total of 125 patients with laboratory-confirmed zoster rash present for < or =72 h were assigned treatment with either 40 mg of sorivudine once daily or 800 mg of acyclovir five times daily, both taken orally for 7 days. Patients were assessed daily until all lesions crusted and then monthly for 6 months for postherpetic neuralgia (PHN) and for 12 months for recurrent or new episodes of zoster. Sorivudine significantly shortened the median period of new vesicle formation from 3.0 to 4.0 days (log rank P = .0001). Sorivudine was effective regardless of duration of rash before treatment. Zoster recurrences and new episodes were experienced by fewer patients assigned sorivudine (11%) than acyclovir (26%, P = .037). No differences were seen in incidence, severity, or duration of either acute neuritis or PHN. Both treatments were well tolerated.
Acute Disease↗
Phenotypic diversity in siblings with partial androgen insensitivity syndrome.
The androgen insensitivity syndrome is a heterogeneous disorder with a wide spectrum of phenotypic abnormalities, ranging from complete female to ambiguous forms that more closely resemble males. The primary abnormality is a defective androgen receptor protein due to a mutation of the androgen receptor gene. This prevents normal androgen action and thus leads to impaired virilisation. A point mutation of the androgen receptor gene affecting two siblings with partial androgen insensitivity syndrome is described. One had cliteromegaly and labial fusion and was raised as a girl, whereas the other sibling had micropenis and penoscrotal hypospadias and was raised as a boy. Both were shown to have the arginine 840 to cysteine mutation. The phenotypic variation in this family is thus dependent on factors other than abnormalities of the androgen receptor gene alone.
Female↗
Sexual relationships, risk behaviour, and condom use in the spread of sexually transmitted infections to heterosexual men.
OBJECTIVE: To examine the effect of patient defined non-regular sexual relationships and other risk behaviours on the incidence of sexually transmitted infections in heterosexual men and the role of condom use in the prevention of their spread. DESIGN: A prospective cross sectional study of sexual behaviour reported by a standardised self administered questionnaire in new patients who presented for screening and diagnosis. SETTING: A genitourinary medicine clinic in west London. SUBJECTS: 957 consecutive newly attending heterosexual men who completed a sexual behaviour questionnaire in 1993/94. MAIN OUTCOME MEASURES: Variables relating to sociodemographic status, sexual behaviour, condom use, sexually transmitted infections and testing for HIV infection, stratified by the reporting of non-regular partners. RESULTS: We found that the 65% of men who reported non-regular sexual partners were more likely to be white collar class (d = 7.5%, 95% CI = 1.3, 13.7) and to have had sexual intercourse with non-United Kingdom born women (d = 7.8%, 95% CI = 3.5, 12.2). They also reported coitarche before 16 years of age (d = 13.4%, 95% CI = 8.0, 18.8) and many more sexual partners both in the last year (d = 13.1%, 95% CI = 10.2, 16.0) and in their lifetime (d = 27.9%, 95% CI = 21.6, 34.2). They were significantly more likely to practise anal intercourse (d = 8.7%, 95% CI = 3.3, 14.1), to smoke (d = 16.3%, 95% CI = 9.8, 22.6), to drink alcohol (d = 4.9%, 95% CI = 1.2, 8.6), and to have chlamydial infection (d = 5.7%, 95% CI = 2.2, 9.2), of which 30% was subclinical. Increasing condom use with regular partners correlated with decreasing incidence of urethral infection (gonorrhoeal and/or chlamydial infection) (p < 0.03) and candidal balanitis (p < 0.03) and a greater likelihood of no infection being detected (p = 0.0002). Use of condoms with non-regular partners was much more frequent than with regular partners (d = 21.4%, 95% CI = 16.7, 26.1). However, we found evidence of oral transmission of urethral gonorrhoea and chlamydial infection among men who reported always using condoms. HIV infection was found in only two men (0.2%), both of whom reported intercourse with non-United Kingdom born women. CONCLUSIONS: Heterosexual men who reported non-regular sexual relationships compensated for their increased risk lifestyle by using condoms more frequently and showed only an increased incidence of chlamydial infection. More consistent condom use with regular partners was significantly associated with the absence of sexually transmitted infection. These findings suggest that transmission between regular partners has been underestimated.
Adolescent↗
Low incidence of androgen receptor gene mutations in human prostatic tumors using single strand conformation polymorphism analysis.
It is possible that structural changes of the androgen receptor (AR) contribute to the insensitivity of prostatic carcinomas to endocrine therapy. We have isolated DNA from 58 human prostate tumor specimens (31 carcinomas pretreatment, 13 carcinomas after relapse to hormonal therapy, and 14 benign prostatic hyperplasia), three established human prostate carcinoma cell lines and two transplantable human prostatic carcinoma xenografts. Twelve pairs of oligonucleotide primers were used to amplify the majority of the coding region of the AR gene and the products screened for mutations using single-strand conformation polymorphism (SSCP) techniques. In one tumor sample a cytosine to guanine transition in exon F which leads to substitution of glutamic acid for the wild type glutamine at position 798 of the ligand binding domain was detected. The same mutation was also found in the patient's genomic DNA and as been described in a patient with partial androgen insensitivity syndrome. Intronic mutations were detected in two of the benign prostatic hyperplasia samples, and a silent mutation at nucleotide 995 was found to be present in eight poorly differentiated carcinomas, one BPH specimen, as well as in the cell line DU145 (18% of the samples studied). In agreement with most of the literature, these studies indicate that AR mutations are rare both prior to therapy and even in androgen relapsed tumors.
Humans↗
Functional analysis of six androgen receptor mutations identified in patients with partial androgen insensitivity syndrome.
Partial androgen insensitivity syndrome (PAIS) is caused by defects in the androgen receptor gene and presents with a wide range of undervirilization phenotypes. We studied the consequences of six androgen receptor ligand-binding domain mutations on receptor function in transfected cells. The mutations, Met742Ile, Met780Ile, Gln798Glu, Arg840Cys, Arg855His and Ile869Met, were identified in PAIS patients with phenotypes representing the full spectrum seen in this condition. In all cases the androgen receptor was found to be defective, suggesting that the mutation is the cause of the clinical phenotype. The Gln798Glu mutation is exceptional in that it did not cause an androgen-binding defect in our system, although the mutant receptor was defective in transactivation assays. This mutation may affect an aspect of binding not tested, or may be part of a functional subdomain of the ligand-binding domain involved in transactivation. Overall we found milder mutations to be associated with milder clinical phenotypes. There is also clear evidence that phenotype is not solely dependent on androgen receptor function. Some of the mutant receptors were able to respond to high doses of androgen in vitro, suggesting that patients carrying these mutations may be the best candidates for androgen therapy. One such mutation is Ile869Met. A patient carrying this mutation has virilized spontaneously at puberty, so in vivo evidence agrees with the experimental result. Thus a more complete understanding of the functional consequences of androgen receptor mutations may provide a more rational basis for gender assignment in PAIS.
Amino Acid Sequence↗
Expression of beta 3-adrenoceptor mRNA in rat tissues.
1. This study examines the expression of beta 3-adrenoceptor messenger RNA (beta 3-AR mRNA) in rat tissues to allow comparison with atypical beta-adrenoceptors determined by functional and radioligand binding techniques. 2. A reverse transcription/polymerase chain reaction protocol has been developed for determining the relative amounts of beta 3-AR mRNA in rat tissues. 3. Measurement of adipsin and uncoupling protein (UCP) mRNA was used to examine all tissues for the presence of white and brown adipose tissue which may contribute beta 3-AR mRNA. 4. The beta 3-AR mRNA is expressed at high levels in brown and white adipose tissue, stomach fundus, the longitudinal/circular smooth muscle of both colon and ileum, and colon submucosa. There was substantial expression of adipsin in colon submucosa and moderate expression in fundus, suggesting that in these regions at least some of the beta 3-AR signal may be contributed by fat. Pylorus and colon mucosa showed moderate levels of beta 3-AR mRNA with lower levels of adipsin. Ileum mucosa and submucosa showed low but readily detectable levels of beta 3-AR. 5. Expression of adipsin in rat skeletal muscles coupled to very low levels of beta 3-AR mRNA indicates that the observed beta 3-AR may be due to the presence of intrinsic fat. beta 3-AR mRNA was virtually undetectable in heart, lung and liver. These results raise the possibility that the atypical beta-AR demonstrated by functional and/or binding studies in muscle and in heart is not the beta 3-AR. 6. By use of two different sets of primers for amplification of beta 3-AR cDNA, no evidence was found for differential splicing of the mRNA in any of the tissues examined. 7. The detection of beta 3-AR mRNA in the gut mucosa and submucosa suggests that in addition to its established roles in lipolysis, thermogenesis and regulation of gut motility beta 3-AR may subserve other functions in the gastrointestinal tract. The absence of beta 3-AR mRNA in rat heart or its presence with adipsin in skeletal muscle suggests that atypical beta-adrenoceptor responses in heart and skeletal muscle are unlikely to be mediated by beta 3-AR.
Animals↗
A new point mutation in the luteinising hormone receptor gene in familial and sporadic male limited precocious puberty: genotype does not always correlate with phenotype.
Genomic DNA from two families with male limited precocious puberty was examined for mutations of the LH receptor gene. In family 1, several members of the pedigree have FMPP, whereas in family 2 there is only one affected subject. A point mutation (T --> C at nucleotide 1192) resulting in substitution of threonine for methionine 398 in the second transmembrane domain of the LH receptor protein was found in both families. In addition, one member of family 1 has the mutation, but no evidence of precocious puberty. All obligate carriers within this family were shown to have the mutation, and it was not detected in 94 chromosomes from unaffected and unrelated white subjects. In family 2, the index case was the only one to have the mutation. A previously unreported neutral dimorphism (C --> T at nucleotide 1065) is also described.
Base Sequence↗
Erythropoietin gene expression in fetal and adult sheep kidney.
The exact location of the cells which express erythropoietin (Epo) in the kidney is still controversial, with conflicting reports suggesting that both peritubular interstitial cells or proximal tubular epithelial cells are possible sites of Epo expression. In the present study we have examined the location of Epo-expressing cells in fetal and adult sheep kidneys after Epo expression was stimulated by anaemia. In situ hybridization histochemistry was performed using synthetic oligonucleotide probes complementary to part of the sheep Epo cDNA sequence or a riboprobe (cRNA) for ovine Epo of 520 bases. Epo expression was confined to peritubular cells of the kidney cortex, in general in the area close to the cortico-medullary junction. In some severely anaemic adult sheep kidneys, Epo-expressing cells were also found in the outer cortex. In addition we located Epo-expressing cells in the kidneys not only of anaemic fetuses (89-140 d of gestation, term = 150d) but also in kidneys from normal fetuses 60-110d of gestation. Again, Epo expression was seen only in peritubular cells of the kidney cortex. These findings confirm that the kidney is an important site of Epo production, in the sheep, from at least 0.4 gestation, but also show that there is no ontogenic change in the cellular site of production within the kidney.
Animals↗