Abstract

Motile cilia, also called flagella, are found across a broad range of species; some cilia propel prokaryotes and eukaryotic cells like sperm, while cilia on epithelial surfaces create complex fluid patterns e.g., in the brain or lung. For sperm, the picture has emerged that the flagellum is not only a motor but also a sensor that detects stimuli from the environment, computing the beat pattern according to the sensory input. Thereby, the flagellum navigates sperm through the complex environment in the female genital tract. However, we know very little about how environmental signals change the flagellar beat and, thereby, the swimming behavior of sperm. It has been proposed that distinct signaling domains in the flagellum control the flagellar beat. However, a detailed analysis has been mainly hampered by the fact that current comprehensive analysis approaches rely on complex microscopy and analysis systems. Thus, knowledge on sperm signaling regulating the flagellar beat is based on custom quantification approaches that are limited to only a few aspects of the beat pattern, do not resolve the kinetics of the entire flagellum, rely on manual, qualitative descriptions, and are only a little comparable among each other. Here, we present SpermQ, a ready-to-use and comprehensive analysis software to quantify sperm motility. SpermQ provides a detailed quantification of the flagellar beat based on common time-lapse images acquired by dark-field or epi-fluorescence microscopy, making SpermQ widely applicable. We envision SpermQ becoming a standard tool in flagellar and motile cilia research that allows to readily link studies on individual signaling components in sperm and distinct flagellar beat patterns.

Highlights

  • Reproduction starts with fertilization, the fusion of sperm and egg

  • We have developed SpermQ, an analysis software that allows to comprehensively study the flagellar beat pattern and sperm steering using microscopy techniques that can be readily applied by almost any lab, i.e., dark-field and epifluorescence microscopy

  • The detected flagellar trace is adjusted to the center of the Gaussian curve, as this defines the precise location of the flagellum in the image (Figure 2D)

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Summary

Introduction

Reproduction starts with fertilization, the fusion of sperm and egg. In mammals, sperm need to pass a complex environment, the female genital tract, to reach the site of fertilization. Sperm need to pass a complex environment, the female genital tract, to reach the site of fertilization On their way through the female genital tract, sperm cells are guided by different physical and chemical cues. Several physical and chemical cues guide the sperm cell through the complex environment in the female genital tract. Chemotaxis, thermotaxis, haptotaxis, and rheotaxis have been described to control the sperm’s swimming path [1]. The flagellum transduces this sensory input from the environment into a distinct flagellar beat pattern [2]. A symmetrical flagellar beat results in a straight swimming path of the sperm cell, whereas asymmetries in the beat pattern lead to a

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