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Transillumination-Assisted Dissection of Specific Stages of the Mouse Seminiferous Epithelial Cycle for Downstream Immunostaining Analyses
Published on: October 7, 2020
Morphological analysis of interstitial cells in murine epididymis using light microscopy and transmission electron
Tasuku Hiroshige1, Kei-Ichiro Uemura2, Shingo Hirashima3
1Division of Microscopic and Developmental Anatomy, Department of Anatomy, Kurume University School of Medicine, Kurume, 830-0011, Japan; Department of Urology, Kurume University School of Medicine, Kurume, 830-0011, Japan.
Abstract:
Smooth muscle contraction of the epididymis plays an important role in sperm transport. Although PDGFRα-positive interstitial cells (PDGFRα (+) ICs) are thought to be involved in controlling smooth muscle movement via intercellular signaling, they have not yet been reported to date in the epididymis. Therefore, we aimed to investigate the morphological characteristics of PDGFRα (+) ICs in the interstitial space of the murine epididymis. Immunohistochemistry showed that PDGFRα (+) ICs co-labeled with CD34 (PDGFRα (+) CD34 (+) ICs were distributed in the interstitial space of the murine epididymis from the initial segment (IS) to the cauda of the epididymis. PDGFRα (+) ICs that were not co-labeled with CD34 (PDGFRα (+) CD34 (-) ICs) were observed just beneath the epithelium from the corpus to the cauda but not in the IS. Both types of PDGFRα (+) ICs were in close proximity to each other as well as the surrounding nerves and macrophages. In addition, PDGFRα (+) CD34 (-) ICs beneath the epithelium were also in close proximity to the basal cells. Using transmission electron microscopy, we identified ICs that possessed elongated and woven cellular processes and were in close proximity to each other, surrounding the cells in the interstitial space. In the murine epididymis, it is suggested that there are two subtypes of ICs that show different distribution patterns depending on the segment, which may reflect segmental differences in mechanisms of sperm transport, forming a cellular network by physical interactions in the murine epididymis.
Insights
This study identifies two subtypes of platelet-derived growth factor receptor alpha-positive interstitial cells (PDGFRα (+) ICs) in the murine epididymis. These cells form a network, potentially influencing sperm transport through segment-specific distribution and interactions.
Area of Science:
- Reproductive biology
- Cell biology
- Histology
Background:
- Smooth muscle contraction in the epididymis is crucial for sperm transport.
- Platelet-derived growth factor receptor alpha-positive interstitial cells (PDGFRα (+) ICs) are implicated in smooth muscle regulation but uncharacterized in the epididymis.
Purpose of the Study:
- To investigate the morphological characteristics and distribution of PDGFRα (+) ICs in the murine epididymis.
- To determine the relationship of PDGFRα (+) ICs with other cell types and structures in the epididymal interstitium.
Main Methods:
- Immunohistochemistry was used to identify and localize PDGFRα (+) ICs, co-labeling with CD34.
- Transmission electron microscopy was employed to examine the ultrastructure of interstitial cells.
Main Results:
- Two subtypes of PDGFRα (+) ICs were identified: PDGFRα (+) CD34 (+) ICs throughout the epididymis and PDGFRα (+) CD34 (-) ICs beneath the epithelium (corpus to cauda).
- Both IC subtypes were found in close proximity to nerves, macrophages, and, in the case of PDGFRα (+) CD34 (-) ICs, basal cells.
- Ultrastructural analysis revealed interstitial cells with elongated, interwoven processes forming a cellular network.
Conclusions:
- The murine epididymis harbors two distinct subtypes of PDGFRα (+) ICs with segment-specific distributions.
- These IC subtypes may contribute to segment-specific sperm transport mechanisms through their network formation and interactions with surrounding cells and nerves.
