Plasticity of basal cells during postnatal development in the rat epididymis

Shum, Winnie; Hill, Eric; Brown, Dennis; Breton, Sylvie

doi:10.1530/rep-12-0510

Cited by 39 publications

(54 citation statements)

References 70 publications

(125 reference statements)

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“…BCs form a homogeneous network in the basal region of all mouse epididymal segments and extend single intraepithelial processes exclusively in the IS. This contrasts with rat BCs, which showed luminal-reaching projections in more distal segments [35,36,40]. Most MPs are located between epithelial cells and the basement membrane, where they form a basolateral network that is intertwined with the network made by BCs.…”

Section: Discussionmentioning

confidence: 78%

“…The expression of KRT5, F4/80, and CD11c, which are used traditionally as markers of BCs, macrophages, and dendritic cells, respectively, has been previously reported in the epididymis [40,42,43,45,48]. EPIDIDYMAL BASAL CELLS ARE NOT IMMUNE CELLS epididymal segments (the narrowest region of the corpus is occasionally absent from the sections), from the most proximal region, the IS, to the caput, the corpus, and the most distal region, the cauda.…”

Section: Bcs and Mps Are Present In All Mouse Epididymal Segmentsmentioning

confidence: 93%

“…For BC identification, we used a polyclonal rabbit anti-KRT5 antibody (Abcam), as we have previously reported that the protein is specific for BCs [40]. A polyclonal rabbit anti-laminin antibody (Sigma-Aldrich) was used to label the basal lamina, which represents a major component of the basement membrane.…”

Section: Antibodies Immunofluorescence and Microscopy Analysesmentioning

confidence: 99%

“…They exhibit segment-specific morphological characteristics, varying from a dome-shaped cell located beneath epithelial cells [34,35] to cells that extend a long and narrow body process projecting into the lumen between adjacent epithelial cells [36]. Epididymal BCs specifically display high expression of cyclooxygenase-1 [36][37][38], claudin-1 [36,39], and keratin (KRT5) [40].…”

Section: Introductionmentioning

confidence: 99%

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Epithelial Basal Cells Are Distinct from Dendritic Cells and Macrophages in the Mouse Epididymis1

Shum

Smith

Cortez-Retamozo

et al. 2014

Biology of Reproduction

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The epithelium that lines the epididymal duct establishes the optimal milieu in which spermatozoa mature, acquire motility, and are stored. This finely tuned environment also protects antigenic sperm against pathogens and autoimmunity, which are potential causes of transient or permanent infertility. The epididymal epithelium is pseudostratified and contains basal cells (BCs) that are located beneath other epithelial cells. Previous studies showed that in the mouse epididymis, BCs possess macrophage-like characteristics. However, we previously identified a dense population of cells belonging to the mononuclear phagocyte (MP) system (comprised of macrophages and dendritic cells) in the basal compartment of the mouse epididymis and showed that a subset of MPs express the macrophage marker F4/80. In the present study, we evaluate the distribution of BCs and MPs in the epididymis of transgenic CD11c-EYFP mice, in which EYFP is expressed exclusively in MPs, using antibodies against the BC marker keratin 5 (KRT5) and the macrophage marker F4/80. Immunofluorescence labeling for laminin, a basement membrane marker, showed that BCs and most MPs are located in the basal region of the epithelium. Confocal microscopy showed that in the initial segment, both BCs and MPs project intraepithelial extensions and establish a very intricate network. Flow cytometry experiments demonstrated that epididymal MPs and BCs are phenotypically distinct. BCs do not express F4/80, and MPs do not express KRT5. Therefore, despite their proximity and some morphological similarities with peritubular macrophages and dendritic cells, BCs do not belong to the MP system.

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Section: Discussionmentioning

confidence: 78%

Section: Bcs and Mps Are Present In All Mouse Epididymal Segmentsmentioning

confidence: 93%

Section: Antibodies Immunofluorescence and Microscopy Analysesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Epithelial Basal Cells Are Distinct from Dendritic Cells and Macrophages in the Mouse Epididymis1

Shum

Smith

Cortez-Retamozo

et al. 2014

Biology of Reproduction

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“…Fibroblasts in the primary cultures were limited by adding 0.1 mg/ml collagenase IV in the culture medium during the first 24 h of culture. The culture medium was changed every day for EEC proliferation for 4 d. The purities of major cell types in the EECs were determined using immunostaining of molecular markers, including aquaporin 9 (for principal cells) (25), keratin 5 (for basal cells) (26), and F4/80 (for macrophages) (27). Triplicate stains were performed for each cell isolation, and at least 200 total cells were spontaneously counted for each staining.…”

Section: Cell Isolationmentioning

confidence: 99%

Pattern Recognition Receptor–Initiated Innate Antiviral Responses in Mouse Epididymal Epithelial Cells

Zhu

Zhao

Liu

et al. 2015

The Journal of Immunology

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Viral infections of the epididymis may impair male fertility and spread sexually transmitted pathogens. The innate antiviral immune responses in the epididymis have yet to be intensively investigated. This study found that mouse epididymal epithelial cells (EECs) constitutively express several viral sensors, including TLR3, retinoic acid–inducible gene I, and DNA-dependent activator of IFN regulatory factors. Other DNA sensors, including p204 and cGMP-AMP synthase, can be induced by transfection of synthetic HSV genomic DNA (HSV60). TLR3 and retinoic acid–inducible gene I in EECs can be activated by their common agonist, polyinosinic-polycytidylic acid [poly(I:C)]. The signaling pathway of DNA sensors can be initiated by HSV60. Both poly(I:C) and HSV60 induced the expression of type 1 IFNs and various antiviral proteins, including IFN-stimulated gene 15, 2′,5′-oligoadenylate synthetase, and myxovirus resistance 1. Poly(I:C), but not HSV60, also dramatically induced the expression of major proinflammatory cytokines, including TNF-α and MCP-1, in EECs. In vivo assay confirmed that the local injection of poly(I:C) or HSV60 induced the innate antiviral responses in EECs. This study provided novel insights into the mechanisms underlying the innate antiviral responses in the mouse epididymis.

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Three‐Dimensional Cell Culture of Epididymal Basal Cells and Organoids: A Novel Tool for Toxicology

Dufresne,

Gregory,

Pinel

et al. 2024

Current Protocols

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Spermatozoa are formed in the testis but must transit through the epididymis to acquire motility and the ability to fertilize. The epididymis is a single convoluted tubule comprising several anatomically and physiologically distinct regions. The pseudostratified epithelium consists of multiple cell types, including principal cells, clear cells, narrow cells, and apical cells, that line the lumen of the epididymis. Basal cells are present at the base of the epithelium, and halo cells, which includes macrophages/monocytes, mononuclear phagocytes, and T lymphocytes, are also present in the epithelium. Several aspects of this complex spermatozoan maturation process are well established, but a great deal remains poorly understood. Given that dysfunction of the epididymis has been associated with male infertility, in vitro tools to study epididymal function and epididymal sperm maturation are required. Our lab and others have previously developed human, rat, and mouse epithelial principal cell lines, which have been used to address certain questions, such as about the regulation of junctional proteins in the epididymis, as well as the toxicity of nonylphenols. Given that the epididymal epithelium comprises multiple cell types, however, a 3D in vitro model provides a more comprehensive and realistic tool that can be used to study and elucidate the multiple aspects of epididymal function. The purpose of this article is to provide detailed information regarding the preparation, maintenance, passaging, and immunofluorescent staining of rat epididymal organoids derived from adult basal cells, which we have demonstrated to be a type of adult stem cell in the rat epididymis. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC.Basic Protocol 1: Isolation of epididymal cellsBasic Protocol 2: Magnetic activated cell sorting and isolation of basal cellsBasic Protocol 3: Preparation and culture of epididymal basal cell organoidsBasic Protocol 4: Passage of epididymal basal cell organoidsBasic Protocol 5: Freezing and thawing of epididymal basal cell organoidsBasic Protocol 6: Immunofluorescent staining of epididymal basal cell organoids

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Plasticity of basal cells during postnatal development in the rat epididymis

Cited by 39 publications

References 70 publications

Epithelial Basal Cells Are Distinct from Dendritic Cells and Macrophages in the Mouse Epididymis1

Epithelial Basal Cells Are Distinct from Dendritic Cells and Macrophages in the Mouse Epididymis1

Pattern Recognition Receptor–Initiated Innate Antiviral Responses in Mouse Epididymal Epithelial Cells

Three‐Dimensional Cell Culture of Epididymal Basal Cells and Organoids: A Novel Tool for Toxicology

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