ATP secretion in the male reproductive tract: essential role of CFTR

Ruan, Ye Chun; Shum, Winnie; Belleannée, Clémence; Silva, Nicolas Da; Breton, Sylvie

doi:10.1113/jphysiol.2012.230581

Cited by 47 publications

(45 citation statements)

References 77 publications

(206 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To do so, the epididymis was perfused at the pH of 5.5 with or without the CFTR inhibitor CFTR inh172 (10 μM) [43,49,50]. As shown in Figure 2, on average, under control condition, the pH of the acidic solution changed from 5.8 to 6.11 ± 0.05 after 10 min and 6.35 ± 0.06 after 20 min (n = 7).…”

Section: Cystic Fibrosis Transmembrane Regulator Participates In the mentioning

confidence: 99%

“…The Cd was perfused in vivo through the lumen, as we have described previously [24,42,43]. Briefly, the distal epididymis was exposed by an abdominal incision under a dissecting microscope and the vas deferens was cannulated using a micro cannula (0.61 mm O. D., 0.28 mm I. D., Warner Instruments, Hamden, CT, USA).…”

Section: In Vivo Microperfusion Of the Mouse CD And Determination Of mentioning

confidence: 99%

See 1 more Smart Citation

Relative contribution of clear cells and principal cells to luminal pH in the mouse epididymis†

Park¹,

Battistone²,

Kim³

et al. 2017

Biology of Reproduction

Self Cite

View full text Add to dashboard Cite

While spermatozoa undergo epididymal maturation, they remain quiescent thanks to the establishment of a low luminal pH. This study is aimed at determining how epithelial cells lining the epididymal lumen work together to maintain and regulate this acidic milieu. In particular, we examined the relative contribution of clear cells (CCs) and principal cells (PCs) to this process. Functional analysis in the mouse cauda epididymidis (Cd) perfused in vivo showed that the pH of a control solution remained constant at pH 6.6 after perfusion through the Cd lumen. In contrast, the pH of both an acidic (pH 5.8) and alkaline (pH 7.8) perfusate was progressively restored toward the control acidic pH. Pharmacological studies indicated the contribution of cystic fibrosis transmembrane regulator, previously shown to be present in the apical membrane of PCs, to the recovery from an acidic pH of 5.8. In addition, we found that CCs and PCs equally contribute to the recovery from an alkaline of 7.8, via the H + pumping vacuolar ATPase (V-ATPase) located in CCs, and the Na + /H + exchanger type 3 (NHE3) located in PCs. Immunofluorescence labeling showed apical membrane accumulation of the V-ATPase in CCs at pH 7.8, and its internalization at pH 5.8 compared to pH 6.6. Immunofluorescence showed expression of NHE3, but absence of NHE2, in PCs located in the Cd. RT-PCR and western blotting showed expression of NHE3 in all epididymal regions. Luminal 8-(4-chlorophenylthio)adenosine 3 ,5 -cyclic monophosphate (cpt-cAMP) partially inhibited luminal pH recovery from pH 7.8. However, cpt-cAMP induced an increase in V-ATPase apical membrane accumulation at this pH. Cell fractionation studies showed the apical accumulation of NHE3 from intracellular vesicles at pH 7.8 versus 6.6, and prevention of this effect by cpt-cAMP. These results indicate the participation of both CCs and PCs in the regulation of luminal pH in the epididymis. Our study also shows the dual role of PCs in HCO 3 − and H + secretion, and that this switch from base to acid secretion depends on the luminal environment. Characterization of the respective roles of CCs and PCs in the regulation of the optimal luminal condition for epididymal sperm maturation should provide new frameworks for the evaluation and treatment of male infertility. Summary SentenceThis study shows the equal contribution of clear and principal cells to proton secretion, and the dual role of principal cells in bicarbonate and proton secretion, depending on luminal cues.

show abstract

Section: Cystic Fibrosis Transmembrane Regulator Participates In the mentioning

confidence: 99%

Section: In Vivo Microperfusion Of the Mouse CD And Determination Of mentioning

confidence: 99%

Relative contribution of clear cells and principal cells to luminal pH in the mouse epididymis†

Park¹,

Battistone²,

Kim³

et al. 2017

Biology of Reproduction

Self Cite

View full text Add to dashboard Cite

show abstract

“…Cells for FACS were further passed through a 35-m filter, and isolation of a clear cell population by FACS was performed at the MGH Flow Cytometry Core facility (Boston, MA). The validity and assessment of contamination of our FACS isolation technique have been shown previously (18,46,59).…”

Section: Methodsmentioning

confidence: 99%

“…EGFP-positive clear cells and EGFP-negative cells were isolated by FACS, either from the entire epididymis of 3 B1-EGFP mice or separately from the caput and cauda epididymis of four mice (46). Total RNA was extracted using RNeasy micro kit (Qiagen, Valencia, CA) following the manufacturer's protocol.…”

Section: Methodsmentioning

confidence: 99%

Circulating aldosterone induces the apical accumulation of the proton pumping V-ATPase and increases proton secretion in clear cells in the caput epididymis

Roy

Hill

Ruan

et al. 2013

American Journal of Physiology-Cell Physiology

Self Cite

View full text Add to dashboard Cite

ϩ -ATPase (V-ATPase) and acidify the lumen of the epididymis, a process that is essential for male fertility. The renin-angiotensin-aldosterone system (RAAS) regulates fluid and electrolyte balance in the epididymis, and a previous study showed binding of aldosterone exclusively to epididymal clear cells (Hinton BT, Keefer DA. Steroid Biochem 23: 231-233, 1985). We examined here the role of aldosterone in the regulation of V-ATPase in the epididymis. RT-PCR showed expression of the mineralocorticoid receptor [MR; nuclear receptor subfamily 3, group C member 2 (NR3C2)] and 11-␤-dehydrogenase isozyme 2 (HSD11␤2) mRNAs specifically in clear cells, isolated by fluorescence-activated cell sorting from B1-enhanced green fluorescent protein (EGFP) mice. Tail vein injection of adult rats with aldosterone, 1,2-dioctanoyl-snglycerol (DOG), or 8-(4-chlorophenylthio)-cAMP (cpt-cAMP) induced V-ATPase apical membrane accumulation and extension of V-ATPaselabeled microvilli in clear cells in the caput epididymis but not in the cauda. V-ATPase activity was measured in EGFP-expressing clear cells using the intracellular pH (pH i)-sensing dye seminaphthorhodafluor-5F-5-(and 6)-carboxylic acid, acetoxymethyl ester acetate (SNARF-5F). Aldosterone induced a rapid increase in the rate of Na ϩ -and bicarbonate-independent pH i recovery following an NH4Cl-induced acid load in clear cells isolated from the caput but not the cauda. This effect was abolished by concanamycin A, spironolactone, and chelerythrine but not myristoylated-protein kinase inhibitor (mPKI) or mifepristone. Thus aldosterone increases V-ATPasedependent proton secretion in clear cells in the caput epididymis via MR/NR3C2 and PKC activation. This study, therefore, identifies aldosterone as an active member of the RAAS for the regulation of luminal acidification in the proximal epididymis. aldosterone; nongenomic; epididymis; clear cell; V-ATPase pump THE EPIDIDYMIS IS AN IMPORTANT male reproductive organ involved in the maturation and storage of spermatozoa. The pseudostratified epithelium of the epididymis is composed of numerous cell types, including principal, basal, and clear cells that function in concert to tightly regulate the luminal environment in which sperm transit (51). One major function of clear cells in the epididymis is to acidify the luminal fluid via the vacuolar proton-pumping H ϩ -ATPase (V-ATPase), which is located on the apical membrane (6,7,13,14). Under resting conditions, a large fraction of V-ATPase resides in a subapical pool of vesicles, and upon activation the V-ATPase is trafficked to the apical membrane. This is accompanied by an increase in apical membrane surface area resulting in the formation and elongation of apical microvillar projections. Thus microvilli elongation directly correlates with V-ATPase activity and has been used as a measure of clear cell activation (6,7,19,39,49). The importance of clear cells to male fertility has been demonstrated, as impairment of V-ATPase activity by genetic manipulation or by environmental factor...

show abstract

“…ATP activates both Ca 2+ and cAMP-dependent Cl − conductances in rat epididymal cells [75]. ATP is released from principal cells in the cauda epididymis of mice and evidence presented that the CFTR is involved in the release mechanism [354]. Since mutations of CFTR are a leading cause of infertility, the authors propose that defective ATP signalling in the epididymis might contribute to the dysfunction.…”

Section: Epididymismentioning

confidence: 99%

Purinergic signalling in the reproductive system in health and disease

Burnstock

2013

Purinergic Signalling

View full text Add to dashboard Cite

There are multiple roles for purinergic signalling in both male and female reproductive organs. ATP, released as a cotransmitter with noradrenaline from sympathetic nerves, contracts smooth muscle via P2X1 receptors in vas deferens, seminal vesicles, prostate and uterus, as well as in blood vessels. Male infertility occurs in P2X1 receptor knockout mice. Both short-and long-term trophic purinergic signalling occurs in reproductive organs. Purinergic signalling is involved in hormone secretion, penile erection, sperm motility and capacitation, and mucous production. Changes in purinoceptor expression occur in pathophysiological conditions, including pre-eclampsia, cancer and pain.

show abstract

ATP secretion in the male reproductive tract: essential role of CFTR

Cited by 47 publications

References 77 publications

Relative contribution of clear cells and principal cells to luminal pH in the mouse epididymis†

Relative contribution of clear cells and principal cells to luminal pH in the mouse epididymis†

Circulating aldosterone induces the apical accumulation of the proton pumping V-ATPase and increases proton secretion in clear cells in the caput epididymis

Purinergic signalling in the reproductive system in health and disease

Contact Info

Product

Resources

About