Changes in Deoxyribonucleoprotein During Spermiogenesis in the Bull: Increased [3h]actinomycin D Binding to Nuclear Chromatin of Morphologically Abnormal Spermatozoa

Abstract

Previously reported work has shown that a reduction in the rate oftritium-labelled actinomycin D ([3H]AMD) binding to spermatids and spermatozoa reflects the extent of the change in nuclear chromatin which occurs during spermiogenesis. Therefore, the kinetics of binding to the deoxyribonucleoprotein (DNP) of live, morphologically abnormal, testicular and ejaculated spermatozoa from five infertile and three control bulls have been studied by the use of quantitative autoradiography in order to examine suspected defects in the nuclear chromatin and particularly in the binding of nuclear protein to DNA.

Quantitative fluorometry of abnormal mouse sperm nuclei

“…In addition to being highly distorted, these nuclei were consistently thicker. This increased thickness may have affected (Gledhill et al, 1971). This was subsequently related to structural defects in the chromatin (Gledhill, 1972).…”

Susceptibility of Sperm Chromatin to Acid Denaturation in Situ: A Study in Endogenous Fsh-Deprived Adult Male Bonnet Monkeys (Macaca radiata)

“…Abnormal alterations in the kinetics of germ cell transformations and defective spermiogenesis have been implicated for the qualitative differences observed in sperm from FSH-deprived monkeys [I, 2, 141. Due to the condensation and compaction that sperm nuclear chromatin undergoes during spermiogenesis and epididymal maturation, the DNA of normal spermatozoa is relatively resistant to denaturation induced either by heat or acid treatment [7,8,211. Susceptibility to such denaturation treatment has been studied by measuring changes in the ratio of emitted red (R) and green (G) fluorescences by sperm nuclei following acridine orange (AO) binding to sperm DNA using either the two-parametric flow cytometric assay [5] or fluorescence microscopy [ 17,241.…”

Distribution of DNA, nuclear micro-heterogeneities and compaction of the chromatin in rabbit epididymal spermatozoa. Ultrastructural evaluation of the Feulgen-like technique using osmium ammine

“…This can be evaluated by measuring the decondensation ability of the chromatin using, for example, swelling of sperm heads by detergents (Rodriguez et al, 1985;Rosenborg ef al, 1990), or quantification of accessible protein side groups such as sulfhydryl groups (Bedford and Calvin, 1974), amino groups , or epitopes of the main sperm nucleoprotein, the protamine (Rodriguez et al, 1990). The accessibility of DNA is also one such criterion (Gledhill et al, 1971 This can be evaluated using, for example, intercalating dyes Kasten, 1960) and has been widely used in light microscopy. In spermatozoa, the amount of Feulgen-positive DNA varies after they are released from the testis, and before they achieve fertilisation; the total DNA content per sperm nucleus remains stable (Esnault and Nicolle, 1976 (Loir and Lanneau, 1984), including HCI which is routinely used during the first step of the Feulgen technique.…”