Allosteric interactions between the oxytocin receptor and the β2-adrenergic receptor in the modulation of ERK1/2 activation are mediated by heterodimerization

The oxytocin receptor (OTR) and the β₂-adrenergic receptor (β₂AR) are key regulators of uterine contraction. These two receptors are targets of tocolytic agents used to inhibit pre-term labor. Our recent study on the nature of OTR- and β₂AR-mediated ERK1/2 activation in human hTERT-C3 myometrial cells suggested the presence of an OTR/β₂AR hetero-oligomeric complex (see companion article). The goal of this study was to investigate potential allosteric interactions between OTR and β₂AR and establish the nature of the interactions between these receptors in myometrial cells. We found that OTR-mediated ERK1/2 activation was attenuated significantly when cells were pretreated with the β₂AR agonist isoproterenol or two antagonists, propranolol or timolol. In contrast, pretreatment of cells with a third β₂AR antagonist, atenolol resulted in an increase in OTR-mediated ERK1/2 activation. Similarly, β₂AR-mediated ERK1/2 activation was strongly attenuated by pretreatment with the OTR antagonists, atosiban and OTA. Physical interactions between OTR and β₂AR were demonstrated using co-immunoprecipitation, bioluminescence resonance energy transfer (BRET) and protein-fragment complementation (PCA) assays in HEK 293 cells, the latter experiments indicating the interactions between the two receptors were direct. Our analyses suggest physical interactions between OTR and β₂AR in the context of a new heterodimer pair lie at the heart of the allosteric effects.     

A Mathematical Model of Uterine Dynamics and its Application to Human Parturition

We have developed a simple mathematical model with three physiologically significant states to describe the changes in intrauterine pressure associated with a contraction during human parturition. The myometrium is modelled as a set of smooth muscle cells, each of which is in one of three states (quiescent, contracted, refractory) at a given time. These states are occupied according to a cycle governed by three temporal parameters. The solutions of the equations describing the model show an oscillatory behavior for particular values of these parameters, which is very similar to the time dependant development of intrauterine pressure during labor. Due to its non-linear terms, our model could lead to chaotic oscillations (in the mathematical sense), whose clinical counterpart may occur in cases of dystocia. Despite its simplicity, this model appears to be a useful guide to further investigations of the oscillatory behavior of the myometrium, or other smooth muscles, in normal and pathological situations.     

剖宫产子宫肌层单层锁边缝合术

目的:评价剖宫产子宫下段横切口缝合技术及子宫肌层单层锁边缝合的可行性。方法:选取我院2008年12月至2010年12月剖宫产病例1920例,940例采用连续加褥式包埋缝合子宫,为对照组;980例采用子宫肌层单层锁边缝合术,为试验组。结果:试验组总手术时间为(28.08±4.15)min,对照组总手术时间为(36.25±5.67)min,两组比较,差异有统计学意义(p<0.01);试验组术后排气时间为(16.10±8.29)h,对照组术后排气时间为(24.26±4.28)h,差异有统计学意义(p<0.05);试验组术中出血量为(210±60)ml,显著低于对照组(280±50)ml,差异有统计学意义(p<0.01);试验组拆线时切口感染率为0.2%,对照组拆线时切口感染率为1%,差异有统计学意义(p<0.05)。而切皮至胎儿娩出时间、新生儿Apgar评分、人均输血量、术后体温一次>38℃、术后三天体温<37℃五项指标比较,两组差异无统计学意义(p>0.05)。结论:子宫肌层单层锁边缝合优于连续加褥式包埋缝合术,进一步提高了产科手术操作技术水平,值得临床推广应用。     

剖宫产子宫肌层单层锁边缝合术

目的：评价剖宫产子宫下段横切口缝合技术及子宫肌层单层锁边缝合的可行性。方法：选取我院2008年12月至2010年12月剖宫产病例1920例,940例采用连续加褥式包埋缝合子宫,为对照组;980例采用子宫肌层单层锁边缝合术,为试验组。结果：试验组总手术时间为（28.08±4.15）min,对照组总手术时间为（36.25±5.67）min,两组比较,差异有统计学意义（p〈0.01）;试验组术后排气时间为（16.10±8.29）h,对照组术后排气时间为（24.26±4.28）h,差异有统计学意义（p〈0.05）;试验组术中出血量为（210±60）ml,显著低于对照组（280±50）ml,差异有统计学意义（p〈0.01）;试验组拆线时切口感染率为0.2%,对照组拆线时切口感染率为1%,差异有统计学意义（p〈0.05）。而切皮至胎儿娩出时间、新生儿Apgar评分、人均输血量、术后体温一次〉38℃、术后三天体温〈37℃五项指标比较,两组差异无统计学意义（p〉0.05）。结论：子宫肌层单层锁边缝合优于连续加褥式包埋缝合术,进一步提高了产科手术操作技术水平,值得临床推广应用。     

Rat myometrial smooth muscle cells show high levels of gap junctional communication under a variety of culture conditions

Summary Gap junctional communciation was examined in rat myometrial smooth muscle cells cultured under a variety of conditions. As a functional measure of gap junctional communication, donor cells were microinjected with the fluorescent dye, Lucifer yellow, and the transfer of dye from donor cells to primary neighbor cells was monitored by fluorescence microscopy. In a myometrial smooth muscle cell line established from midgestation (Day 10) rats, high levels of dye transfer, in excess of 90%, were observed in primary cultures and at Passages 1 and 10. A slight decrease in dye transfer to 75% was observed at Passage 5. Similarly, high levels of dye transfer were observed in a smooth muscle cell line established from the myometrium of a late-gestation (Day 19) rat under subconfluent as well as confluent culture conditions. Myometrial smooth muscle cell cultures established from sexually immature 19-day-old rats also exhibited high levels of dye transfer in primary cultures and at Passage 10. Treatment of primary myometrial smooth muscle cell cultures derived from immature 19-day-old rats with 17β-estradiol (50 ng/ml) and 4-pregnen-3,20-dione (150 ng/ml) for 48 h in vitro had no significant effect on the high levels of dye transfer. Thus, extensive dye transfer was observed in the rat myometrial smooth muscle cells under all culture conditions examined, regardless of sexual maturity or gestational stage of the animal, in vitro hormone treatment, or cell density.     

Active metabolism of arachidonic acid by Kaposi sarcoma cells cultured from lung biopsies (KS-3); identification by HPLC and MS/MS of the predominant metaboilte secreted as the 11,12-epoxy-eicosatrienoic acid

The development of long-term culture of AIDS-KS cells has allowed us to investigate further a possible vascular origin of Kaposi sarcoma. Taking into account the relative specificity of arachidonic acid (AA) metabolism according to cell type, the AA ‘cascade’ was analyzed in cultured KS-3 cells established from lung biopsies and compared to human umbilical venous endothelial (H-UVE) cells and human myometrial smooth muscle (H-MSM) cells, under basal conditions and after stimulation with vasoactive agents such as histamine or thrombin. Considering strictly the ‘prostaglandin’ profile given by RIAs, the metabolism of AA was closer, whilst not identical, to H-UVE than to H-MSM cells. However, evaluation of all the eicosanoids released from [³H]AA labeled KS-3 cells revealed that the predominant metabolite was not prostacyclin (PGI₂), as suggested from PG RIAs, but an epoxy-eicosatrienoic acid (EET), identified as the 11, 12 isomer by HPLC and MS/MS. The synthesis of this EET is probably cytochrome P-450 monooxygenase dependent. Its potential role in the development of the KS tumor cells is under investigation.     

2-Methoxyethanol inhibits gap junctional communication in rat myometrial myocytes

The glycol ethers 2-methoxyethanol (2-ME) and 2-ethoxyethanol (2-EE) prolong gestation in rodents. Because gap junctions in the myometrium likely facilitate parturition, the present study examined inhibition of gap junctional communication by 2-ME and 2-EE in myometrial smooth-muscle cell cultures. To measure gap junctional communication, the fluorescent dye Lucifer yellow was injected into cultured cells and the transfer of the dye to adjacent cells was scored with epifluorescence microscopy. The data are presented as the percentage of cells adjacent to the microinjected cell that exhibited dye following microinjection. A 30 min treatment with 32 or 63 mmol/L 2-ME decreased dye transfer to 71% and 63%, respectively (p0.05; control 90%). Similarly, 2-EE inhibited dye transfer, although myometrial cells were less sensitive to 2-EE compared to 2-ME. Dye transfer returned to control levels after 2 h in the continued presence of 2-ME. The primary metabolite of 2-ME, methoxyacetic acid (MAA), had no effect on dye transfer at concentrations equimolar to 2-ME. Because 2-ME and 2-EE inhibited gap junctional communication only at high concentrations and because the inhibition reversed in the continued presence of the compounds, it is suggested that glycol ethers delay parturition by a mechanism independent of a direct action on myometrial gap junctions.