Fine structure of the nucleus during spermiogenesis

Abstract

Several investigators have reported microfibrils in chromosomes of various plants and animals. Fibrils about 100 Å thick have been found in all chromosomes studied. Often these are associated in pairs and form fibrils 200–250 Å thick. By dissolving calf thymus chromosomes isolated in saline-versene of p_H 6.5 in water we have obtained a nucleoprotein in the form of fibrils about 100 Å thick and identical in appearance to the fibrils found in sections through osmium fixed chromosomes. We assume therefore that the microfibrils making up the chromosomes represent the nucleoprotein. If calf thymus chromosomes are left in saline-versene of p_H 8 before dissolving in water the non-histone protein autolyzes and is lost from the fibrous nucleoprotein. In electron micrographs the 100 Å fibrils are now seen to consist of two 40 Å fibrils which seem to correspond to the nucleohistone. — During spermiogenesis the non-histone protein generally disappears from the nucleus. Electronmicroscopic studies of spermiogenesis in a large number of species show that at this time the 100 Å fibrils become visibly double, consisting of two 40 Å fibrils. In sperm where dichroism and X-ray diffraction indicate orientation of DNA, these fibrils are oriented in the same way. No other structures are visible in the sperm nucleus and in the mature sperm these fibrils fill the nucleus solidly. We conclude that the microfibrils correspond to the DNA with associated basic protein and that the DNA molecule is arranged in the long axis of the microfibrils. Late in spermiogenesis the 40 A fibrils associate into large units. In some species progressively thicker bundles are formed which may be oriented in the long axis of the sperm or irregularly twisted through the nucleus.