Anat Cell Biol.  2017 Dec;50(4):301-305. 10.5115/acb.2017.50.4.301.

Application of stereological methods for unbiased estimation of sperm morphology in the mice induced by busulfan

• Panahi S
• Abdollahifar MA
• Aliaghaei
• Nazarian H
• Paktinat S
• Abdi S
• Farahani R

• KMID: 2399899
• DOI: http://doi.org/10.5115/acb.2017.50.4.301

Abstract

Busulfan is an anticancer drug, which causes the apoptosis germ cells and azoospermia in humans and animals. Abnormal morphology of spermatozoa related to the male infertility. The sperm morphology is evaluation of sperm size, shape and appearance characteristics should be assessed by carefully observing a stained sperm sample under the microscope. Evaluation of sperm morphology has been considered as one of the most important factors for a successful fertilization and determining sperm quality. The mice were assigned to tow experimental groups: control and busulfan. Each group included six mice that were housed under standard conditions. The volume was estimated using the nucleator method. The sperm's flegellum and mid-piece length was estimated by counting the number of intersections between the tails and Merz grid test line in an unbiased counting frame, superimposed on live images of sperms. Our results demonstrated a significant different in the volume and surface of the sperm's head and the length of the sperm's flagellum in the control and busulfan groups. Busulfan can effect on the volume of the sperm's head and the length of the sperm's flagellum in rat.

Keyword

Busulfan; Sperm morphology; Stereology

MeSH Terms

Animals
Apoptosis
Azoospermia
Busulfan*
Fertilization
Flagella
Germ Cells
Head
Humans
Infertility, Male
Male
Methods*
Mice*
Rats
Spermatozoa*
Tail
Busulfan

Figure

• 

  Fig. 1 Microphotograph illustrating morphology of sperm in control (A) and busulfan (B) group in mice (Diff-Quik staining, ×40).

• 

  Fig. 2 Estimation of the sperms mid-piece and flagellum length. (A) Four cycloids were located at a rectangle. The length of each cycloid was equal to twice the length of its minor axis (r). The area associated with the cycloids was calculated by multiplying the X by Y and divided by the length of the four cycloids to achieve the area per length. The total number of the intersections between the sperms mid-piece and flagellum axes and the cycloid were counted. The cycloid was positioned parallel to the vertical axis (×100). (B) Schematic representation of the structure of a spermatozoon (×100).

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