Calcium-releasing activity induced by nuclei of mouse fertilized early embryos

At fertilization,repectitive transient rises of intracellular calcium concentration occur in all mammals studied so far .It has been shown that calcium rises could be induced when mouse fertilized 1-,2-cell nuclei were trans-planted into unfertilized eggs and that the reconstituted embryo could be activated .Howerver,whecther the capability of inducing calcium rises occurs in all stages of mammalian embryos remains unknown ,In this study ,by using the nuclear transplantation technique and measurement of intracellular calcium rises in living cells,we showed that only the nuclei from mouse fertilized 1-cell and 2-cell embryos ,neither the nuclei from 4-,8-cell and ethanol activated parthe-nogenetic embryos nor 2 or 3 nuclei of electrofused 4-cell stage syncytium ,have calcium -releasing activity when they were transferred into unfertilized mature oocytes,Our results indicate that the calcium-releasing activity in nuclei of 1-,2-cell embryos is produced during fertilization and exists at the special stage of fertilized early embryos.These sug-gested that the capacity of inducting calcium release activity in fertilized early embryos is important for normal embryonic development.     

金鱼精子头在卵细胞质中转化的超微结构研究

用透射电镜研究了金鱼卵子受精后起始阶段精子头在卵细胞质中的转化过程，结果表明：这种转化同样必须经精子核膜的崩解、原来高度致密的精子染色质的解聚与分散以及新的核膜（雄原核核膜）的形成。     

Exceptional sperm cooperation in the wood mouse

Spermatozoa from a single male will compete for fertilization of ova with spermatozoa from another male when present in the female reproductive tract at the same time. Close genetic relatedness predisposes individuals towards altruism, and as haploid germ cells of an ejaculate will have genotypic similarity of 50%, it is predicted that spermatozoa may display cooperation and altruism to gain an advantage when inter-male sperm competition is intense. We report here the probable altruistic behaviour of spermatozoa in an eutherian mammal. Spermatozoa of the common wood mouse, Apodemus sylvaticus, displayed a unique morphological transformation resulting in cooperation in distinctive aggregations or 'trains' of hundreds or thousands of cells, which significantly increased sperm progressive motility. Eventual dispersal of sperm trains was associated with most of the spermatozoa undergoing a premature acrosome reaction. Cells undergoing an acrosome reaction in aggregations remote from the egg are altruistic in that they help sperm transport to the egg but compromise their own fertilizing ability.     

First cleavage plane of the mouse egg is not predetermined but defined by the topology of the two apposing pronuclei

Studies of experimentally manipulated embryos have led to the long-held conclusion that the polarity of the mouse embryo remains undetermined until the blastocyst stage. However, recent studies reporting that the embryonic-abembryonic axis of the blastocyst arises perpendicular to the first cleavage plane, and hence to the animal-vegetal axis of the zygote, have led to the claim that the axis of the mouse embryo is already specified in the egg. Here we show that there is no specification of the axis in the egg. Time-lapse recordings show that the second polar body does not mark a stationary animal pole, but instead, in half of the embryos, moves towards a first cleavage plane. The first cleavage plane coincides with the plane defined by the two apposing pronuclei once they have moved to the centre of the egg. Pronuclear transfer experiments confirm that the first cleavage plane is not determined in early interphase but rather is specified by the newly formed topology of the two pronuclei. The microtubule networks that allow mixing of parental chromosomes before dividing into two may be involved in these processes.     

酶法制备立体专一的同位素标记化合物

本文讨论了酶法制备立体专一的氢同位素标记化合物的六种方法。同时对碳、氮、氧、磷、硫同位素标记化合物的酶法制备也作了简要叙述。     

The immunoglobulin superfamily protein Izumo is required for sperm to fuse with eggs

Representing the 60 trillion cells that build a human body, a sperm and an egg meet, recognize each other, and fuse to form a new generation of life. The factors involved in this important membrane fusion event, fertilization, have been sought for a long time. Recently, CD9 on the egg membrane was found to be essential for fusion, but sperm-related fusion factors remain unknown. Here, by using a fusion-inhibiting monoclonal antibody and gene cloning, we identify a mouse sperm fusion-related antigen and show that the antigen is a novel immunoglobulin superfamily protein. We have termed the gene Izumo and produced a gene-disrupted mouse line. Izumo-/- mice were healthy but males were sterile. They produced normal-looking sperm that bound to and penetrated the zona pellucida but were incapable of fusing with eggs. Human sperm also contain Izumo and addition of the antibody against human Izumo left the sperm unable to fuse with zona-free hamster eggs.     

Role for sperm in spatial patterning of the early mouse embryo

Despite an apparent lack of determinants that specify cell fate, spatial patterning of the mouse embryo is evident early in development. The axis of the post-implantation egg cylinder can be traced back to organization of the pre-implantation blastocyst. This in turn reflects the organization of the cleavage-stage embryo and the animal-vegetal axis of the zygote. These findings suggest that the cleavage pattern of normal development may be involved in specifying the future embryonic axis; however, how and when this pattern becomes established is unclear. In many animal eggs, the sperm entry position provides a cue for embryonic patterning, but until now no such role has been found in mammals. Here we show that the sperm entry position predicts the plane of initial cleavage of the mouse egg and can define embryonic and abembryonic halves of the future blastocyst. In addition, the cell inheriting the sperm entry position acquires a division advantage and tends to cleave ahead of its sister. As cell identity reflects the timing of the early cleavages, these events together shape the blastocyst whose organization will become translated into axial patterning after implantation. We present a model for axial development that accommodates these findings with the regulative nature of mouse embryos.     

In vitro production of functional sperm in cultured neonatal mouse testes

Spermatogenesis is one of the most complex and longest processes of sequential cell proliferation and differentiation in the body, taking more than a month from spermatogonial stem cells, through meiosis, to sperm formation. The whole process, therefore, has never been reproduced in vitro in mammals, nor in any other species with a very few exceptions in some particular types of fish. Here we show that neonatal mouse testes which contain only gonocytes or primitive spermatogonia as germ cells can produce spermatids and sperm in vitro with serum-free culture media. Spermatogenesis was maintained over 2?months in tissue fragments positioned at the gas-liquid interphase. The obtained spermatids and sperm resulted in healthy and reproductively competent offspring through microinsemination. In addition, neonatal testis tissues were cryopreserved and, after thawing, showed complete spermatogenesis in vitro. Our organ culture method could be applicable through further refinements to a variety of mammalian species, which will serve as a platform for future clinical application as well as mechanistic understanding of spermatogenesis.     

Development of reconstituted mouse eggs suggests imprinting of the genome during gametogenesis

It has been suggested that the failure of parthenogenetic mouse embryos to develop to term is primarily due to their aberrant cytoplasm and homozygosity leading to the expression of recessive lethal genes. The reported birth of homozygous gynogenetic (male pronucleus removed from egg after fertilization) mice and of animals following transplantation of nuclei from parthenogenetic embryos to enucleated fertilized eggs, is indicative of abnormal cytoplasm and not an abnormal genotype of the activated eggs. However, we and others have been unable to obtain such homozygous mice. We investigated this problem further by using reconstituted heterozygous eggs, with haploid parthenogenetic eggs as recipients for a male or female pronucleus. We report here that the eggs which receive a male pronucleus develop to term but those with two female pronuclei develop only poorly after implantation. Therefore, the cytoplasm of activated eggs is fully competent to support development to term but not if the genome is entirely of maternal origin. We propose that specific imprinting of the genome occurs during gametogenesis so that the presence of both a male and a female pronucleus is essential in an egg for full-term development. The paternal imprinting of the genome appears necessary for the normal development of the extraembryonic membranes and the trophoblast.     

地高辛标记cRNA探针原位杂交检测大鼠脑NMDAR—L mRNA

采用地高辛（Ｄｉｇｏｘｉｎ Ｄｉｇ）标记神经递质受体ＮＭＤＡＲ－Ｌ ｃＲＮＡ探针技术进行原位杂交,研究了Ｗｉｓｔａｒ新生大鼠脑神经细胞ＮＭＤＡＲ－Ｌ的基因表达。光学显微镜观察结果显示,大脑皮层、海马等区有明显的ＮＭＤＡＲ－Ｌ ｍＲＮＡ表达,胞浆着蓝色,胞核不着色,背景浅谈,反差显著。实验结果表明地高辛标记ｃＲＮＡ探针能快速、准确地检测出ＮＭＤＡＲ－Ｌ ｍＲＮＡ,为进一步研究这一拟受体在脑中表达和分