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Updated: Aug 24, 2026
A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Published on: February 6, 2018
Morphological changes of mouse spermatozoa in uterus as revealed by scanning and transmission electron microscopy
Abstract:
Submicroscopic alterations occurring in mouse spermatozoa during their sojourn into the female reproductive tract--as revealed by Scanning Electron Microscopy and Transmission Electron Microscopy using ruthenium red as marked for glycosaminoglycans--are described in the present investigation. The results show that the surface of the plasma membrane covering the acrosomal region of the spermatozoa found in the uterus--at different intervals of time after copulation (between 10 minutes up to 20 hrs)--are provided with an increasing number of granules and small vesicles. Some of the surface structures appear gradually leading to perforation of discrete areas of plasma membrane in regions corresponding to acrosomal areas covering the head of spermatozoa. Similarly ruthenium red preparations, show a significant decrease of glycosaminoglycans and their sialic acid moieties, which specifically precede further vesiculation and perforation of the sperm plasma membrane covering the same acrosomal regions.
Insights
Mouse sperm undergo significant surface changes in the female reproductive tract. Alterations include increased granules, vesiculation, and plasma membrane perforation, linked to glycosaminoglycan changes.
Area of Science:
- Reproductive Biology
- Cell Biology
- Spermatozoa Ultrastructure
Background:
- Spermatozoa undergo critical transformations after ejaculation to achieve fertilization capability.
- The female reproductive tract environment influences sperm surface modifications.
- Understanding these changes is key to reproductive success.
Purpose of the Study:
- To investigate submicroscopic alterations in mouse spermatozoa within the female reproductive tract.
- To identify changes in sperm plasma membrane and glycosaminoglycans post-copulation.
Main Methods:
- Utilized Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM).
- Employed ruthenium red staining to visualize glycosaminoglycans.
- Examined spermatozoa at various time points after copulation (10 minutes to 20 hours).
Main Results:
- Spermatozoa in the uterus exhibited increasing granules and vesicles on the acrosomal region plasma membrane.
- Progressive vesiculation and perforation of the plasma membrane were observed.
- Ruthenium red staining revealed a significant decrease in glycosaminoglycans and sialic acid moieties preceding membrane alterations.
Conclusions:
- Spermatozoa undergo dynamic ultrastructural changes in the female reproductive tract.
- These modifications involve plasma membrane vesiculation and perforation, influenced by glycosaminoglycan alterations.
- The findings provide insights into sperm capacitation and readiness for fertilization.
