Culture and purification of SD rat corpus cavernosum endothelial cells by enzymatic digestion combined with mechanical extrusion and fixed‐point digestion

Andrologia ◽  
2021 ◽  
Author(s):  
Ying Chen ◽  
Tao Qi ◽  
Shu‐Guang Zhu ◽  
Hao Li ◽  
Jia‐Xin Feng ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Fixed Point ◽  
Corpus Cavernosum ◽  
Enzymatic Digestion ◽  
Sd Rat

scholarly journals Transcriptional profiling of human cavernosal endothelial cells reveals distinctive cell adhesion phenotype and role for claudin 11 in vascular barrier function

2009 ◽  
Vol 39 (2) ◽  
pp. 100-108 ◽  
Author(s):  
Hunter Wessells ◽  
Chris J. Sullivan ◽  
Yoshiaki Tsubota ◽  
Karen L. Engel ◽  
Bryan Kim ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Barrier Function ◽  
Umbilical Vein ◽  
Transcriptional Profiling ◽  
Corpus Cavernosum ◽  
Focal Adhesions ◽  
Receptor Interaction ◽  
Molecular Features ◽  
Junction Protein

To determine specific molecular features of endothelial cells (ECs) relevant to the physiological process of penile erection we compared gene expression of human EC derived from corpus cavernosum of men with and without erectile dysfunction (HCCEC) to coronary artery (HCAEC) and umbilical vein (HUVEC) using Affymetrix GeneChip microarrays and GeneSifter software. Genes differentially expressed across samples were partitioned around medoids to identify genes with highest expression in HCCEC. A total of 190 genes/transcripts were highly expressed only in HCCEC. Gene Ontology classification indicated cavernosal enrichment in genes related to cell adhesion, extracellular matrix, pattern specification and organogenesis. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed high expression of genes relating to ECM-receptor interaction, focal adhesions, and cytokine-cytokine receptor interaction. Real-time PCR confirmed expression differences in cadherins 2 and 11, claudin 11 (CLDN11), desmoplakin, and versican. CLDN11, a component of tight junctions not previously described in ECs, was highly expressed only in HCCEC and its knockdown by siRNA significantly reduced transendothelial electrical resistance in HCCEC. Overall, cavernosal ECs exhibited a transcriptional profile encoding matrix and adhesion proteins that regulate structural and functional characteristics of blood vessels. Contribution of the tight junction protein CLDN11 to barrier function in endothelial cells is novel and may reflect hemodynamic requirements of the corpus cavernosum.

Separation, culture and identification of SD rat corpus cavernosal endothelial cells

Andrologia ◽  
2012 ◽  
Vol 44 (4) ◽  
pp. 250-255 ◽  
Author(s):  
J. Chen ◽  
C. L. Sun ◽  
Z. Chen ◽  
H. J. Xiao ◽  
T. Qi ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Sd Rat

Cytotoxicity of different intracavernous vasoactive drugs on cultured endothelial cells of human corpus cavernosum penis

Urology ◽  
2004 ◽  
Vol 64 (3) ◽  
pp. 598-602 ◽  
Author(s):  
Dirk Schultheiss ◽  
Adrian Pilatz ◽  
Alexander I. Gabouev ◽  
Norbert Schlote ◽  
Jörg Wefer ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Corpus Cavernosum ◽  
Vasoactive Drugs ◽  
Cultured Endothelial Cells

scholarly journals Efficient isolation and identification of primary endothelial cells from bovine aorta by collagenase P

2020 ◽  
Vol 6 (2) ◽  
pp. e15-e15
Author(s):  
Faezeh Seif ◽  
Alireza Kheirollah ◽  
Hossein Babaahmadi-Rezaei
Keyword(s):  
Endothelial Cells ◽  
Enzymatic Digestion ◽  
Gene Expressions ◽  
Isolation And Identification ◽  
Digestion Method ◽  
Aortic Endothelial Cells ◽  
Endothelin 1 ◽  
Carboxy Terminal ◽  
Endothelial Nitric Oxide ◽  
Aortic Endothelial

Introduction: Endothelial cells are widely used among researchers for investigation of cardiovascular diseases, particularly atherosclerosis. Since aortic endothelial cells are able to be cultured in high passages, these cells are suitable for physiological and pathological studies of blood vessels. Objectives: The aim of this study was employing a digestion method to isolate the endothelial cells from bovine aorta by utilizing collagenase P and, establishing and characterizing isolated primary endothelial cells (IPECs) cultures. Materials and Methods: IPECs were isolated from fresh bovine aorta via enzymatic digestion method using collagenase P. Cell morphology and its functions were assessed by measuring the gene expression of endothelial nitric-oxide synthase (eNOS) and endothelin-1. In order to validate them as IPECs, they were compared with bovine aortic endothelial cells (BAECs) and vascular smooth muscle cells (VSMCs). Effects of synthetic endothelin-1 (100 nm) were assessed on the phosphorylation of Smad2 transcription factor via western blotting in IPECs for periods of one to four hours. Results: In this study, the morphology of IPECs from bovine aorta was identical to that of the BAECs. The gene expressions of endothelin-1 and eNOS were higher than those of BAECs and VSMCs. In addition, synthetic endothelin-1 resulted in the time-dependent boost of phosphorylation of carboxy-terminal Smad2 in the IPECs for periods of two and four hours. Conclusion: The results of this study confirm the efficacy of the enzymatic digestion method in isolating a large number of endothelial cells with morphological and functional characteristics.

Penile erection requires association of soluble guanylyl cyclase with endothelial caveolin-1 in rat corpus cavernosum

2006 ◽  
Vol 290 (5) ◽  
pp. R1302-R1308 ◽  
Author(s):  
A. Elizabeth Linder ◽  
Romulo Leite ◽  
Kimberly Lauria ◽  
Thomas M. Mills ◽  
R. Clinton Webb
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Guanylyl Cyclase ◽  
Penile Erection ◽  
Muscle Relaxation ◽  
Corpus Cavernosum ◽  
Soluble Guanylyl Cyclase ◽  
Smooth Muscle Relaxation ◽  
Membrane Cholesterol ◽  
Caveolin 1

Erectile dysfunction is caused by a variety of pathogenic factors, particularly impaired formation and action of nitric oxide (NO). NO released from nerve endings and corpus cavernosum endothelial cells plays a crucial role in initiating and maintaining increased intracavernous pressure, penile vasodilatation, and penile erection. Classically, these effects are dependent on cGMP synthesized during activation of soluble guanylyl cyclase (sGC) by NO in smooth muscle cells. The enzyme NO synthase in endothelial cells has been localized to caveolae, small invaginations of the plasma membrane rich in cholesterol. Membrane cholesterol depletion impairs acetylcholine-induced relaxation in arteries attributed to an alteration in caveolar structure. It has been shown that sGC may be activated in endothelial caveolae contributing to vasodilation. We hypothesized that caveolae are the platform for sGC/cGMP signaling in cavernosum smooth muscle eliciting erection. Methyl-β-cyclodextrin, a pharmacological tool to deplete membrane cholesterol and disassemble caveolae, impaired rat erectile responses in vivo and cavernosum smooth muscle relaxation induced by the NO donor sodium nitroprusside and the sGC activator 3-(5′-hydroxymethyl-2′-furyl)-1-benzyl indazole in vitro. Methyl-β-cyclodextrin had no effect on cavernosum smooth muscle relaxation induced by NO released upon nerve stimulation or by exogenous cGMP. Furthermore, sGC and caveolin-1, the major coat protein of caveolae, were colocalized in rat corpus cavernosum sinusoidal endothelium. Electron microscopy indicated caveolae disruption in corpus cavernosum treated with methyl-β-cyclodextrin. In summary, our results provide evidence of compartmentalization of sGC in the caveolae of cavernosal endothelial cells contributing to NO signaling mediating smooth muscle relaxation and erection.

scholarly journals Immunohistochemical detection of tyrosine hydroxylase in human corpus cavernosum endothelial cells

2020 ◽  
Vol 19 ◽  
pp. e1071-e1072
Author(s):  
L.E Ximenes ◽  
V. Barbosa De Souza ◽  
J. Britto-Junior ◽  
A. Fregonesi ◽  
R. Destro Saade ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Tyrosine Hydroxylase ◽  
Corpus Cavernosum ◽  
Immunohistochemical Detection

Effects of androgen on extracellular vesicles from endothelial cells in rat penile corpus cavernosum

Andrology ◽  
2021 ◽  
Author(s):  
Ling‐Tao Yan ◽  
Zhi‐Hui Yang ◽  
Haocheng Lin ◽  
Jun Jiang ◽  
Rui Jiang
Keyword(s):  
Endothelial Cells ◽  
Extracellular Vesicles ◽  
Corpus Cavernosum

Endothelin: localization, synthesis, activity, and receptor types in human penile corpus cavernosum

1991 ◽  
Vol 261 (4) ◽  
pp. H1078-H1085 ◽  
Author(s):  
I. Saenz de Tejada ◽  
M. P. Carson ◽  
A. de las Morenas ◽  
I. Goldstein ◽  
A. M. Traish
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Cultured Cells ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Corpus Cavernosum ◽  
High Affinity ◽  
Cells In Culture ◽  
Receptor Sites ◽  
Competition Binding

The localization, synthesis, and activity of endothelin and the receptor types mediating its effects in penile corpus cavernosum were investigated in whole tissue and in cultured cells derived from this tissue. With immunocytochemistry, utilizing an antiendothelin 1 (ET-1) monoclonal antibody, endothelin-like immunoreactivity was localized intensely in the endothelium and to a lesser degree in the trabecular smooth muscle. Human corpus cavernosum endothelial cells in culture expressed preproendothelin 1 mRNA, as determined by Northern blot analysis. Significant amounts of endothelin-like immunoreactivity were measured by radioimmunoassay in the supernatants of corpus cavernosum endothelial cells in culture. Endothelins are potent constrictors and caused long-lasting contractions of corporeal strips in organ chambers. Equilibrium binding analysis of endothelins to their receptor sites revealed high-affinity, specific, and saturable binding of labeled endothelins to corporeal membranes. Competition binding experiments demonstrated receptors with high affinity for ET-1 and -2 and low affinity for ET-3 and another, less abundant, set of receptors with high affinity for ET-1, -2, and -3. Affinity labeling of endothelins to corporeal membranes, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, revealed that ET-1 and -2 cross-linked specifically to three different molecular mass components (75, 52, and 34 kDa). ET-3 bound only to the 34-kDa component. It is concluded that human corpus cavernosum endothelium has the ability to synthesize and release endothelin, that endothelins contract corporeal smooth muscle, and that at least two distinct endothelin receptors may exist and are differentiated by their affinity for ET-3.

scholarly journals A Simple and Nonenzymatic Method to Isolate Human Corpus Cavernosum Endothelial Cells and Pericytes for the Study of Erectile Dysfunction

2020 ◽  
Vol 38 (1) ◽  
pp. 123 ◽  
Author(s):  
Guo Nan Yin ◽  
Jiyeon Ock ◽  
Min-Ji Choi ◽  
Kang-Moon Song ◽  
Kalyan Ghatak ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Erectile Dysfunction ◽  
Corpus Cavernosum
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