Stimulating Effects of Dopamine on Chloride Transport Across the Rat Caudal Epididymal Epithelium in Culture1

Du, Jianyang; Zuo, Wu-Lin; Ruan, Ye Chun; Yang, Zihuan; Chen, Minhui; Chen, Siliang; Li, Sheng; Wu, Zhuangzhi; Xiang, Hui; Zhou, Wen‐Liang

doi:10.1095/biolreprod.108.068346

Cited by 10 publications

(11 citation statements)

References 39 publications

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“…The procedures of tracheal epithelium primary culture have been described previously [44]. In brief, Sprague-Dawley rats weighing 180 g –200 g were killed by CO 2 inhalation.…”

Section: Methodsmentioning

confidence: 99%

Cellular Mechanism Underlying Formaldehyde-Stimulated Cl− Secretion in Rat Airway Epithelium

et al. 2013

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BackgroundRecent studies suggest that formaldehyde (FA) could be synthesized endogeneously and transient receptor potential (TRP) channel might be the sensor of FA. However, the physiological significance is still unclear.Methodology/Principal FindingsThe present study investigated the FA induced epithelial Cl- secretion by activation of TRPV-1 channel located in the nerve ending fiber. Exogenously applied FA induced an increase of I SC in intact rat trachea tissue but not in the primary cultured epithelial cells. Western blot and immunofluorescence analysis identified TRPV-1 expression in rat tracheal nerve ending. Capsazepine (CAZ), a TRPV-1 specific antagonist significantly blocked the I SC induced by FA. The TRPV-1 agonist capsaicin (Cap) induced an increase of I SC, which was similar to the I SC induced by FA. L-703606, an NK-1 specific inhibitor and propranolol, an adrenalin β receptor inhibitor significantly abolished the I SC induced by FA or Cap. In the ion substitute analysis, FA could not induce I SC in the absence of extracelluar Cl-. The I SC induced by FA could be blocked by the non-specific Cl- channel inhibitor DPC and the CFTR specific inhibitor CFTRi-172, but not by the Ca2+-activated Cl- channel inhibitor DIDS. Furthermore, both forskolin, an agonist of adenylate cyclase (AC) and MDL-12330A, an antagonist of AC could block FA-induced I SC.ConclusionOur results suggest that FA-induced epithelial I SC response is mediated by nerve, involving the activation of TRPV-1 and release of adrenalin as well as substance P.

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“…The procedures of tracheal epithelium primary culture have been described previously [44]. In brief, Sprague-Dawley rats weighing 180 g –200 g were killed by CO 2 inhalation.…”

Section: Methodsmentioning

confidence: 99%

Cellular Mechanism Underlying Formaldehyde-Stimulated Cl− Secretion in Rat Airway Epithelium

et al. 2013

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“…As an in vitro model, primary cell culture was utilized for epididymal function studies, such as investigations on the interaction between epididymal epithelial cells and sperm in vitro [2]–[5]. However, primary culture of epididymal epithelial cells generally display limited proliferation potential, and the cells easily lose the functional characteristics during the extended culture [6], [7]. For an in vivo approach, many methods of DNA transfer for tissue could be employed, due to the short experiment period and convenience.…”

Section: Introductionmentioning

confidence: 99%

Trehalose Maintains Vitality of Mouse Epididymal Epithelial Cells and Mediates Gene Transfer

Shen

et al. 2014

PLoS ONE

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In the present study, trehalose was utilized to improve primary culture of mouse epididymal epithelial cells in vitro, and to enhance naked DNA delivery in epididymis in vivo. During the six-day culture, the proliferation activity of the cells in the medium with addition of trehalose was higher than that of those cells cultured in absence of trehalose (p<0.01). To determine the optimal concentration for cell proliferation, a series of trehalose concentrations (0, 60, 120, 180 mM) were tested, and the result indicated that the cell in the medium with 120 mM trehalose showed the highest proliferation potential. The epididymis epithelial cells were cultured in the medium containing 120 mM trehalose upon 16th passage, and they continued expressing markers of epididymal epithelial cell, such as rE-RABP, AR and ER-beta. Our study also indicated that trehalose concentrations of 120–240 mM, especially 180 mM, could effectively enhance DNA delivery into the mouse epididymis epithelial cell in vitro. Moreover, trehalose could induce in vivo expression of exogenous DNA in epididymal epithelial cells and help to internalize plasmid into sperm,which did not influence motility of sperm when the mixture of trehalose (180 mM) and DNA was injected into epididymal lumen through efferent tubule. This study suggested that trehalose, as an effective and safer reagent, could be employed potentially to maintain vitality of mouse epididymal epthetial cells during long-term culture in vitro and to mediate in vitro and in vivo gene transfer.

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“…In addition, pre‐treating the cells with the chloride channel inhibitor DPC at 1 mM (Figure C), which was usually used as nonselective chloride channel blocker (Du et al, ) or with the CFTR channel inhibitor, CFTRinh‐ 172 at 20 µM (Figure D), the Isc induced by G1 was largely inhibited. In contrast the Ca 2+ ‐dependent Cl − channel (CaCC) inhibitor, DIDS (100 µM) (Du et al, ), had no effect on G1 induced short current reaction. These data suggested that the CaCC was not involved and the CFTR was mainly responsible for the anion secretion.…”

Section: Resultsmentioning

confidence: 99%

Functional expression of G protein‐coupled receptor 30 in immature rat epididymal epithelium

Cao

Huang

Zhang

et al. 2016

Cell Biology International

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The aim of this study is to investigate the functional role of G protein-coupled receptor 30 (GPR30) in the epididymis. We found that GPR30 is expressed in the epithelium of the immature rat epididymis and is involved in chloride secretion into the caudal epididymis lumen. The short-circuit current (Isc) experiments showed that in primary cultured caudal epididymis epithelium, activation of GPR30 by its specific agonist G1 induced a mono-phasic current increase, and G15, the specific antagonist of GPR30, could completely inhibit the current induced by G1. The G1-induced Isc was largely blocked by application of the non-specific chloride channel inhibitor diphenylamine-dicarboxylic acid (DPC), or by the cystic fibrosis transmembrane conductance regulator (CFTR) inhibitor CFTR , suggesting that the current was mainly mediated through CFTR. In addition, after stimulating GPR30 by G1, the intracellular concentration of cAMP in the epithelium was significantly increased, indicating that the cAMP signal pathway is involved and could be responsible for the CFTR activation. Finally, to further investigate the function of GPR30 in vivo, G15 was administrated into rats subcutaneously. The osmotic pressure of the micro perfusion solution from epididymis was measured and the sperms were collected. Results showed that there was an osmotic pressure increase of the perfusion solution from G15 treated rats. When the GPR30 was inhibited by G15 endogenously, the motility of sperms decreased. Our data demonstrated that GPR30 is involved in the formation of caudal epididymis fluid micro-environment thus affecting sperm motility.

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Stimulating Effects of Dopamine on Chloride Transport Across the Rat Caudal Epididymal Epithelium in Culture1

Cited by 10 publications

References 39 publications

Cellular Mechanism Underlying Formaldehyde-Stimulated Cl− Secretion in Rat Airway Epithelium

Cellular Mechanism Underlying Formaldehyde-Stimulated Cl− Secretion in Rat Airway Epithelium

Trehalose Maintains Vitality of Mouse Epididymal Epithelial Cells and Mediates Gene Transfer

Functional expression of G protein‐coupled receptor 30 in immature rat epididymal epithelium

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