Presence and Passage Dependent Loss of Biochemical M3 Muscarinic Receptor Function in Human Detrusor Cultured Smooth Muscle Cells

Boselli, Cinzia; Govoni, Stefano; D, Vicini; Lanni, Cristina; Racchi, Marco; D׳Agostino, Gianluigi

doi:10.1016/s0022-5347(05)64242-5

Cited by 9 publications

(5 citation statements)

References 14 publications

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“…Contrary to what has been observed for muscarinic (Boselli et al, 2002) or tachykinin receptors (unpublished observations), the present data indicate that isolated smooth muscle cells from human detrusor specimens express both B 1 and B 2 kinin receptors which are retained despite subculture conditions. Radioligand binding experiments (saturation curves obtained with tritiated agonists) with membranes prepared from cultured cells, indicate a receptor density comparable to that measured in other human cell types, such as fibroblasts (WI38, Meini et al, 1999;HLF-1, Cucchi et al, 2005), or smooth muscle cells (Schmidlin et al, 1998).…”

Section: Discussioncontrasting

confidence: 99%

“…Smooth muscle cells from human bladder have been previously used to verify the effect of different culture conditions on cell growth (Baskin et al, 1993;Lai et al, 2002) or to characterize muscarinic or histamine receptors (Boselli et al, 2002;Neuhaus et al, 2006). The aim of the present study was to evaluate the pharmacological and coupling characteristics of kinin receptors in human detrusor smooth muscle cells, and to study the effect of long-term culture conditions or of proinflammatory cytokines (IL-1b and TNFa) on the expression of kinin receptors, revealed through radioligand binding of agonists.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of kinin receptors in human cultured detrusor smooth muscle cells

Bellucci¹,

Cucchi²,

Santicioli³

et al. 2007

British J Pharmacology

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Background and purpose: Kinins have an important role in inflammatory cystitis and in animal pathophysiological models, by acting on epithelium, fibroblasts, sensory innervation and smooth muscle. The aim of this study was to characterize the receptors responsible for direct motor responses induced by kinins on human detrusor. ]-Lys-BK was 10% of BK). BK also induced prostaglandin E 2 release (EC 50 2.3 nM), whereas no response was detected with the B 1 receptor agonist. The incubation of detrusor smooth muscle cells with interleukin 1b (IL-1b) or tumour necrosis factor-a (TNFa) (10 ng ml À1 ) induced a time-dependent increase in radioligand-specific binding, which was greater for the B 1 than for the B 2 receptor. Conclusions and Implications: Human detrusor smooth muscle cells in culture retain kinin receptors, and represent a suitable model to investigate the mechanisms and changes that occur under chronic inflammatory conditions.

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Section: Discussioncontrasting

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Characterization of kinin receptors in human cultured detrusor smooth muscle cells

Bellucci¹,

Cucchi²,

Santicioli³

et al. 2007

British J Pharmacology

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“…), as well as a decrease in InsP 3 production (Boselli et al . ). Our results also showed a significant drop in the levels of SMA expressed after 1 week in culture, though clear SMA stress fibres were still apparent in the majority of cells.…”

Section: Discussionmentioning

confidence: 97%

The transition of smooth muscle cells from a contractile to a migratory, phagocytic phenotype: direct demonstration of phenotypic modulation

Sandison

Dempster

McCarron

2016

The Journal of Physiology

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Key points Smooth muscle cell (SMC) phenotypic conversion from a contractile to a migratory phenotype is proposed to underlie cardiovascular disease but its contribution to vascular remodelling and even its existence have recently been questioned.Tracking the fate of individual SMCs is difficult as no specific markers of migratory SMCs exist.This study used a novel, prolonged time‐lapse imaging approach to continuously track the behaviour of unambiguously identified, fully differentiated SMCs.In response to serum, highly‐elongated, contractile SMCs initially rounded up, before spreading and migrating and these migratory cells displayed clear phagocytic activity.This study provides a direct demonstration of the transition of fully contractile SMCs to a non‐contractile, migratory phenotype with phagocytic capacity that may act as a macrophage‐like cell. AbstractAtherosclerotic plaques are populated with smooth muscle cells (SMCs) and macrophages. SMCs are thought to accumulate in plaques because fully differentiated, contractile SMCs reprogramme into a ‘synthetic’ migratory phenotype, so‐called phenotypic modulation, whilst plaque macrophages are thought to derive from blood‐borne myeloid cells. Recently, these views have been challenged, with reports that SMC phenotypic modulation may not occur during vascular remodelling and that plaque macrophages may not be of haematopoietic origin. Following the fate of SMCs is complicated by the lack of specific markers for the migratory phenotype and direct demonstrations of phenotypic modulation are lacking. Therefore, we employed long‐term, high‐resolution, time‐lapse microscopy to track the fate of unambiguously identified, fully‐differentiated, contractile SMCs in response to the growth factors present in serum. Phenotypic modulation was clearly observed. The highly elongated, contractile SMCs initially rounded up, for 1–3 days, before spreading outwards. Once spread, the SMCs became motile and displayed dynamic cell‐cell communication behaviours. Significantly, they also displayed clear evidence of phagocytic activity. This macrophage‐like behaviour was confirmed by their internalisation of 1 μm fluorescent latex beads. However, migratory SMCs did not uptake acetylated low‐density lipoprotein or express the classic macrophage marker CD68. These results directly demonstrate that SMCs may rapidly undergo phenotypic modulation and develop phagocytic capabilities. Resident SMCs may provide a potential source of macrophages in vascular remodelling.

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“…In agreement with reports on human BSMCs (Howard et al, 2005), this inhibition of DNA synthesis in rat BSMC was not reflected by reductions in cell number. Although a discrepancy between reduced DNA synthesis and unaltered cell number may be surprising, this is most likely related to the very slow doubling of BSMCs (Malkowicz et al, 1995;Kropp et al, 1999;Boselli et al, 2002), which makes it technically difficult to detect reduced cell numbers unless cells are observed for very long time courses. The inhibition of DNA synthesis may reflect a TGF␤-induced cell cycle arrest, which has been observed in many other cell types (Ewen et al, 1993;Reddy and Howe, 1993;Reynisdóttir et al, 1995;Wiesmann et al, 2000;Seay et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Role of Transforming Growth Factor β in Rat Bladder Smooth Muscle Cell Proliferation

Barendrecht

Mulders²,

Poel³

et al. 2007

J Pharmacol Exp Ther

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Conditions associated with hypertrophy of the urinary bladder have repeatedly been associated with an increased urinary excretion of transforming growth factor (TGF) ␤ in both rats and patients. Because TGF␤ can have both growth-promoting and -inhibiting effects, we have studied its effects on cell growth and death in primary cultures of rat bladder smooth muscle cells. TGF␤1, TGF␤2, or TGF␤3 did not cause apoptosis, but all three isoforms inhibited DNA synthesis with similar potency (EC 50 of approximately 0.1 ng/ml) and efficacy. Such inhibition was antagonized by a specific TGF␤ receptor antagonist and independent of the presence of serum. Mitogen-activated protein kinases (MAPKs) are involved in the control of cell growth, and all three TGF␤ isoforms inhibited activation of the extracellular signal-regulated kinase, c-Jun NH 2 -terminal kinase, and p38 MAPK subfamilies. Nevertheless, the inhibitory effects of the TGF␤ isoforms on DNA synthesis were not affected by presence of inhibitors of the three MAPK pathways. TGF␤ did not alter cell size as measured by flow cytometry or mitochondrial activity, an integrated measure of cell size and number. We conclude that our data do not support the hypothesis that TGF␤ is a mediator of rat bladder hypertrophy.Urinary bladder outlet obstruction (BOO) is a frequent consequence of benign prostatic enlargement or urethral strictures. It leads to an increased bladder size and micturition disorders (Andersson, 2003), the former involving both hypertrophy and hyperplasia at the cellular level. However, the mediators and molecular mechanisms leading to bladder hypertrophy have remained elusive. Studies in rats (Chul Kim et al., 2001) and patients with BOO (MacRae Dell et al., 2000;Monga et al., 2001) have found an increased urinary excretion of transforming growth factor (TGF) ␤. Hence, TGF␤ has been proposed to play a role in the pathophysiology of BOO and bladder hypertrophy.TGF␤ is a pluripotent growth factor that has been implicated in a variety of physiological processes such as proliferation, apoptosis, phenotypic switching, differentiation, and specification of developmental fate (Massagué, 2000). Interestingly, TGF␤ can have differential or even opposite effects depending on the tissue or cell type under investigation (Roberts and Sporn, 1993). For example, TGF␤ can increase the proliferation of airway smooth muscle cells (Chen and Khalil, 2006), but it potently inhibits proliferation in vascular smooth muscle cells due to a G 0 /G 1 arrest via the p38 pathway (Seay et al., 2005). In the heart, TGF␤ can induce cardiomyocyte hypertrophy, whereas cardiac fibroblasts respond with differentiation into myofibroblasts and synthesis of collagen Watkins et al., 2006). Therefore, it remains largely unclear whether increased TGF␤ excretion in experimental and clinical BOO reflects a role as mediator of cell growth, as part of a negative feedback loop limiting cell growth, or as a by-product that at best can be used as a biomarker.The heterogeneity of TGF␤ responses may...

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Presence and Passage Dependent Loss of Biochemical M3 Muscarinic Receptor Function in Human Detrusor Cultured Smooth Muscle Cells

Cited by 9 publications

References 14 publications

Characterization of kinin receptors in human cultured detrusor smooth muscle cells

Characterization of kinin receptors in human cultured detrusor smooth muscle cells

The transition of smooth muscle cells from a contractile to a migratory, phagocytic phenotype: direct demonstration of phenotypic modulation

Role of Transforming Growth Factor β in Rat Bladder Smooth Muscle Cell Proliferation

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