Functional heterogeneity of PDGFRα (+) cells in spontaneously active urogenital tissues

Hashitani, Hikaru; Mitsui, Retsu; Lang, Richard J

doi:10.1002/nau.24431

Cited by 10 publications

(9 citation statements)

References 25 publications

(52 reference statements)

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“…ASMCs are most abundant in the proximal RP, at the PKJ ( Figure 1A – C ), where peristaltic contractions and electrical pacemaker activity originates 5 , 7 , 8 , 9 , 12 , 13 , 14 , 16 , 17 , 18 , 19 , 20 , 21 , 22 . A recently published study by this group identified platelet-derived growth factor receptor-alpha (PDGFRα), in combination with smooth muscle myosin heavy chain (smMHC), as a unique biomarker for these interstitial cells (ICs) 24 , a finding that has been corroborated by other groups 25 . Based on their morphology and protein expression pattern, these cells were called PDGFRα + IC type 1 (PIC1) 24 , 26 .…”

Section: Introductionmentioning

confidence: 78%

Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning

Grainger

Sanders

Drumm

2021

JoVE

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The renal pelvis (RP) is a funnel-shaped, smooth muscle structure that facilitates normal urine transport from the kidney to the ureter by regular, propulsive contractions. Regular RP contractions rely on pacemaker activity, which originates from the most proximal region of the RP at the pelvis-kidney junction (PKJ). Due to the difficulty in accessing and preserving intact preparations of the PKJ, most investigations on RP pacemaking have focused on single-cell electrophysiology and Ca 2+ imaging experiments. Although important revelations on RP pacemaking have emerged from such work, these experiments have several intrinsic limitations, including the inability to accurately determine cellular identity in mixed suspensions and the lack of in situ imaging of RP pacemaker activity. These factors have resulted in a limited understanding of the mechanisms that underlie normal rhythmic RP contractions. In this paper, a protocol is described to prepare intact segments of mouse PKJ using a vibratome sectioning technique. By combining this approach with mice expressing cell-specific reporters and genetically encoded Ca 2+ indicators, investigators may be able to more accurately study the specific cell types and mechanisms responsible for peristaltic RP contractions that are vital for normal urine transport.

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Section: Introductionmentioning

confidence: 78%

Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning

Grainger

Sanders

Drumm

2021

JoVE

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“…ASMCs are most abundant in the proximal RP, at the PKJ (Figure 1A-C), where peristaltic contractions and electrical pacemaker activity originates 5 , 7 , 8 , 9 , 12 , 13 , 14 , 16 , 17 , 18 , 19 , 20 , 21 , 22 . A recently published study by this group identified platelet-derived growth factor receptor-alpha (PDGFRα), in combination with smooth muscle myosin heavy chain (smMHC), as a unique biomarker for these interstitial cells (ICs) 24 , a finding that has been corroborated by other groups 25 . Based on their morphology and protein expression pattern, these cells were called PDGFRα + IC type 1 (PIC1) 24 , 26 .…”

Section: Introductionmentioning

confidence: 78%

Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning

Grainger

Sanders

Drumm

2021

JoVE

View full text Add to dashboard Cite

The renal pelvis (RP) is a funnel-shaped, smooth muscle structure that facilitates normal urine transport from the kidney to the ureter by regular, propulsive contractions.Regular RP contractions rely on pacemaker activity, which originates from the most proximal region of the RP at the pelvis-kidney junction (PKJ). Due to the difficulty in accessing and preserving intact preparations of the PKJ, most investigations on RP pacemaking have focused on single-cell electrophysiology and Ca 2+ imaging experiments. Although important revelations on RP pacemaking have emerged from such work, these experiments have several intrinsic limitations, including the inability to accurately determine cellular identity in mixed suspensions and the lack of in situ imaging of RP pacemaker activity. These factors have resulted in a limited understanding of the mechanisms that underlie normal rhythmic RP contractions. In this paper, a protocol is described to prepare intact segments of mouse PKJ using a vibratome sectioning technique. By combining this approach with mice expressing cell-specific reporters and genetically encoded Ca 2+ indicators, investigators may be able to more accurately study the specific cell types and mechanisms responsible for peristaltic RP contractions that are vital for normal urine transport.

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“…Indeed, functional heterogeneity of PDGFRα + cells is becoming evident in various urogenital organs. In the mouse bladder, stimulation of P2Y 1 receptors induces Ca 2+ transients in PDGFRα + interstitial cells, which did not suppress the LVDCC-dependent spontaneous Ca 2+ transients in the neighbouring detrusor SMCs (Hayashi et al 2019;Hashitani et al 2020). In the renal pelvis, spontaneously active PDGFRα + ASMCs depolarise the membrane of adjacent typical SMCs to induce action potential generation suggesting that PDGFRα + ASMCs act as the pacemaker cells for pyeloureteric peristalsis (Grainger et al 2020;Hashitani et al 2020).…”

Section: Asynchronous Spontaneous Ca 2+ Transients and Stds In Becs O...mentioning

confidence: 89%

“…2020; Hashitani et al . 2020). Therefore, it is of great interest to know if PDGFRα + cells in the SV mucosa may also develop depolarising signals and function as pacemaker cells for SPCs of SV smooth muscle.…”

Section: Introductionmentioning

confidence: 99%

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PDGFRα⁺ subepithelial interstitial cells act as a pacemaker to drive smooth muscle of the guinea pig seminal vesicle

Takeya

Higashi

Hashitani

et al. 2022

The Journal of Physiology

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Smooth muscle cells (SMCs) of the guinea pig seminal vesicle (SV) develop spontaneous phasic contractions, Ca2+ flashes and electrical slow waves in a mucosa‐dependent manner, and thus it was envisaged that pacemaker cells reside in the mucosa. Here, we aimed to identify the pacemaker cells in SV mucosa using intracellular microelectrode and fluorescence Ca2+ imaging techniques. Morphological characteristics of the mucosal pacemaker cells were also investigated using focused ion beam/scanning electron microscopy tomography and fluorescence immunohistochemistry. Two populations of mucosal cells developed spontaneous Ca2+ transients and electrical activity, namely basal epithelial cells (BECs) and subepithelial interstitial cells (SICs). Pancytokeratin‐immunoreactive BECs were located on the apical side of the basement membrane (BM) and generated asynchronous, irregular spontaneous Ca2+ transients and spontaneous transient depolarisations (STDs). The spontaneous Ca2+ transients and STDs were not diminished by 10 μM nifedipine but abolished by 10 μM cyclopiazonic acid (CPA). Platelet‐derived growth factor receptor α (PDGFRα)‐immunoreactive SICs were distributed just beneath the basal side of the BM and developed synchronous Ca2+ oscillations and electrical slow waves, which were suppressed by 3 μM nifedipine and abolished by 10 μM CPA. In SV mucosal preparations in which some smooth muscle bundles remained attached, SICs and residual SMCs developed temporally correlated spontaneous Ca2+ transients. Neurobiotin injected into SICs spread not only to neighbouring SICs but also to neighbouring SMCs or vice versa. These results suggest that PDGFRα+ SICs electrotonically drive the spontaneous contractions of SV smooth muscle. Key points In many visceral smooth muscle organs, spontaneous contractions are electrically driven by non‐muscular pacemaker cells. In guinea pig seminal vesicles (SVs), as yet unidentified mucosal cells appear to drive neighbouring smooth muscle cells (SMCs). Two populations of spontaneously active cells are distributed in the SV mucosa. Basal epithelial cells (BECs) generate asynchronous, irregular spontaneous Ca2+ transients and spontaneous transient depolarisations (STDs). In contrast, subepithelial interstitial cells (SICs) develop synchronous Ca2+ oscillations and electrical slow waves. Pancytokeratin‐immunoreactive (IR) BECs are located on the apical side of the basement membrane (BM), while platelet‐derived growth factor receptor α (PDGFRα)‐IR SICs are located on the basal side of the BM. Spontaneous Ca2+ transients in SICs are synchronised with those in SV SMCs. Dye‐coupling between SICs and SMCs suggests that SICs act as pacemaker cells to drive the spontaneous contractions of SV smooth muscle.

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Functional heterogeneity of PDGFRα (+) cells in spontaneously active urogenital tissues

Cited by 10 publications

References 25 publications

Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning

Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning

Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning

PDGFRα⁺ subepithelial interstitial cells act as a pacemaker to drive smooth muscle of the guinea pig seminal vesicle

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Functional heterogeneity of PDGFRα (+) cells in spontaneously active urogenital tissues

Cited by 10 publications

References 25 publications

Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning

Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning

Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning

PDGFRα+ subepithelial interstitial cells act as a pacemaker to drive smooth muscle of the guinea pig seminal vesicle

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PDGFRα⁺ subepithelial interstitial cells act as a pacemaker to drive smooth muscle of the guinea pig seminal vesicle