Partial illustration of human sperm DNA via microscopy and quantitative analysis of nucleotides

Abstract

AbstractBackgroundgeneral structure of human sperm has not been profiled yet. Human sperm DNA characterization should progress the medical diagnostic and therapeutic methods rather than developing biological sciences. The aim of the present study was to provide biological insights into the common structure of human sperm. The value of this investigation is establishing an initial basic map of sperm head structure that leads to further advanced standardization of normality in this creature. For this purpose, analytical and microscopic methods were applied.MethodsHigh-performance Liquid Chromatography (HPLC) and flow cytometry were hired to quantify the DNA compositions. As well fluorescent, confocal and advanced light microscopy was applied to identify the stained sperm DNA by chromomycinA3 (CMA3) and 5-methylcytosine antibody (5-mc)ResultsHPLC demonstrated the mean values of nucleotide bases’ percentage in the structure of the sperm DNA regardless of the fraction that sperm was collected from gradient wash, sequenced from 27.6%, 8.92%, 27.05% and 35.36%. Also, quantitative flow cytometry of global 5-methylcytosine showed not a regular fluctuation in individuals with normal sperm while, there is a permanent increase in 50% fraction collected from percoll gradients.CMA3-positivity levels as well, were negatively correlated with sperm quality harvest by percoll gradients (p<0.0001), and positively correlated (P<0.05) with global methylation as determined by flow cytometry. Interestingly, in this text microscopy of immunocytochemistry of sperm cells stained by CMA3, demonstrated a different view from cells’ heads.Conclusionsobviously these explorations suggest some new possibilities in assessment of rough chemical level of nucleotides and cytochemistry of sperm head structure. The chromatin brightness presented with CMA3 by microscopy shows a direct relation with more extensive DNA methylation in sperms collected from low gradients of percoll wash. While, fluctuated 5-methylcytosine levels show personal presentation and even exclusive to individual sperm expression. This study induces further research on new assumptions in nuclear equilibrium in the axiom of DNA ladder in related to 5-mcytosine level in human sperm.