The first multi-flagellate spermatozoa in the animal kingdom, discovered in Mastotermes darwiniensis.

In this paper the spermatozoa of Mastotermes darwiniensis is described. It is the first example of multiflagellate animal sperm cell, consisting of a conical head and about hundred flagella. Other remarkable features are the absence of acrosome, the centrioles made up by doublets instead of triplets and the axonemes devoid of central tubules and with doublets bearing only one arm. The flagella are feebly motile.     

A sperm GPI-anchored protein elicits sperm-cumulus cross-talk leading to the acrosome reaction

The acrosome reaction has long been thought to be induced by the zona pellucida. Here we report the identification and function of a novel human sperm glycosylphosphatidylinositol (GPI)-anchored membrane protein, NYD-SP8. The release of the protein during sperm-egg interaction and its binding to the cumulus, the first layer of egg investment, elicits cross-talk between the gametes and produces calcium dependant release of progesterone, which lead to the acrosome reaction. An in vivo mouse model of NYD-SP8 immunization is also established showing a reduced fertility rate. Thus, contrary to accepted dogma, our study demonstrates for the first time that, prior to reaching the zona pellucida, sperm may release a surface protein that acts on the cumulus cells leading to the acrosome reaction, which may be important for determining the outcome of fertilization. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Received 11 August 2008; received after revision 18 December 2008; accepted 22 December 2008     

Localization of vitelline-coat lysin purified from testis of a top shell,Turbo cornutus

Summary The vitelline-coat lysin purified from the testis ofTurbo cornutus was found, by an immunofluorescence technique, to be located in the acrosome of the sperm, which suggested that the lysin reacts with the vitelline-coat in an early phase of fertilization to allow the sperm to penetrate through the coat.We are grateful to Professor M. Akino (Tokyo Metropolitan University) for his helpful advice.     

The viability of pollen grains of a lily (Lilium auratum) and the eggs of the brine-shrimp (Artemia salina) soaked in organic solvents for 10 years

Summary Pollen grains ofLilium auratum which had been stored in n-pentanol, n-butanol and n-propanol for 10 years at –10°C germinated, and the generative nucleus in the pollen tube divided into 2 sperm nuclei. The resting eggs ofArtemia salina soaked in these solvents for 10 years hatched at a high rate.     

Acrosomal hydrolases in buffalo spermatozoa.

Buffalo sperm acrosome resembles its counterpart in other species, being rich in hydrolytic enzymes. Of the enzyme activities estimated, acid phosphatase, beta-N-acetylglucosaminidase and hyaluronidase were low compared to those of ram semen. However, the aryl sulphatase activity was high. GOT activity estimated in sperm preparation may not be of acrosomal origin.     

Gene expression in spermiogenesis

Germ cells convey parental genes to the next generation, and only germ cells perform meiosis, which is a mechanism that preserves the parental genes. The fusion of the products of germ cell meiosis, the haploid sperm and egg, creates the next generation. Sperm are the haploid germ cells that contribute genes to the egg. In preparation for this, the haploid round spermatids produced by meiosis undergo drastic morphological changes to become sperm. During this process of spermiogenesis, the nuclear form of the haploid germ cell takes shape, the mitochondria are rearranged in a specific manner, the flagellum develops and the acrosome forms. Spermatogenesis is supported by precise and orderly regulation of gene expression during the changes in chromatin structure, when protamine replaces histone. In this report, we summarize the molecular mechanisms involved in spermiogenesis.Received 2 September 2004; received after revision 7 October 2004; accepted 7 October 2004     

Two-step acquisition of motility by insect spermatozoa

High motility of eupyrene sperm of a grasshopper (Omocestus ventralis) was induced by cAMP. Both trypsin and cAMP were necessary for high motility of eupyrene sperm of three other species of grasshoppers. The same was found for apyrene sperm of the silkmoth,Bombyx mori, when they had been wahsed free from seminal plasma. On the other hand, in locusts (Locusta migratoria), in which the yellow gland has high Arg-C endopeptidase activity, sperm motility was induced by trypsin, like that of apyrene sperm of Lepidoptera. Thus in these two orders of Insecta, sperm motility appears to be induced by the same two-step process: the first step by Arg-C endopeptidase, and the second by cAMP.     

Hormonal facilitation in the release of sperm from the spermatheca of the red-spotted newt

Several neurotransmitters and hormones with potential to trigger a simultaneous contraction of the oviducts and the spermathecal myoepithelium were examined. Saline (0.05 ml), or 0.05 ml saline plus acetylcholine (9 mg), norepinephrine (50 micrograms), arginine-vasotocin (25 units), prostaglandin F2 alpha (3 micrograms) were injected into the spermathecal region of female newts (n = 24 per group). The numbers of sperm present in the cloacae of prostaglandin-injected animals (107 +/- 30 SEM) were significantly greater than the numbers detected in saline (27 +/- 5 SEM) and in uninjected (14 +/- 3 SEM) controls. Smaller and less consistent increases in the numbers of sperm were detected in the vasotocin- and norepinephrine-injected groups. Study of sections from ovulating female newts failed to produce evidence that pressure from the passage of ova through the posterior portion of the oviduct forced sperm from the spermatheca. Observations indicate an active role for the spermathecal myoepithelium in the discharge of stored sperm and of a role for prostaglandin F2 alpha in triggering that discharge.     

Primary visual centers in the frog (Rana esculenta L.) and the problem of ipsilateral projections. Radioautographic study]

The topographic organization of the frog's primary visual system has been reexamined in Rana esculenta using autoradiographic technique. 10 to 24 hrs following an intraocular injection of tritiated proline, an intense labelling was observed contralaterally in the various retinal projection centres (preoptic area); neuropils of Bellonci; corpus geniculatum of the thalamus; posterior thalamus; posterodorsal preoptic area; magnocellular pretectal nucleus and basal optic nucleus; tectal laminae A to G of Potter), as already described. Furthermore, the presence of a ipsilateral projection within the hypothalamus (preoptic area), the thalamus (neuropils of Bellonci and corpus geniculatum) and the pretectium (posterior thalami neuropil, pretectal optic area) is confirmed. On the other hand, the existence of ipsilateral retinotectal (to Potter's lamina F) and retino-tegmentomescencephalic projections (to the basal optic nucleus), which were thusfar questioned, has been demonstrated.     

Acrosomal hydrolases in buffalo spermatozoa

Summary Buffalo sperm acrosome resembles its counterpart in other species, being rich in hydrolytic enzymes. Of the enzyme activities estimated, acid phosphatase, -N-acetylglucosaminidase and hyaluronidase were low compared to those of ram semen. However, the aryl sulphatase activity was high. GOT activity estimated in sperm preparation may not be of acrosomal origin.NDRI publication No. 75-129.We should like to thank Dr.D. Sundaresan, the Director for encouragement and the Indian Council of Agricultural Research, New Delhi for partially funding this work. One of the authors (PSC) was recipient of a UGC junior fellowship.     

Centrioles: active players or passengers during mitosis?

Centrioles are cylinders made of nine microtubule (MT) triplets present in many eukaryotes. Early studies, where centrosomes were seen at the poles of the mitotic spindle led to their coining as “the organ for cell division”. However, a variety of subsequent observational and functional studies showed that centrosomes might not always be essential for mitosis. Here we review the arguments in this debate. We describe the centriole structure and its distribution in the eukaryotic tree of life and clarify its role in the organization of the centrosome and cilia, with an historical perspective. An important aspect of the debate addressed in this review is how centrioles are inherited and the role of the spindle in this process. In particular, germline inheritance of centrosomes, such as their de novo formation in parthenogenetic species, poses many interesting questions. We finish by discussing the most likely functions of centrioles and laying out new research avenues.     

Sex-peptides: seminal peptides of the <Emphasis Type="Italic">Drosophila</Emphasis> male

Mating affects the reproductive behaviour of insect females: the egg-laying rate increases and courting males are rejected. These post-mating responses are induced mainly by seminal fluid. In Drosophila melanogaster, males transfer two peptides (sex-peptides, = Sps) that reduce receptivity and elicit increased egg laying in their mating partners. Similarities in the open reading frames of the genes suggest that they have arisen by gene duplication. In females, Sps bind to specific sites in the central and peripheral nervous system, and to the genital tract. The binding proteins of the nervous system and genital tract are membrane proteins, but they differ molecularly. The former protein is proposed to be a receptor located at the top of a signalling cascade leading to the two post-mating responses, whereas the latter is a carrier protein moving Sps from the genital tract into the haemolymph. Sps bind to sperm. Together with sperm they are responsible for the persistence of the two post-mating responses. But Sps are the molecular basis of the sperm effect; sperm is merely the carrier.Received 10 February 2003; received after revision 25 April 2003; accepted 1 May 2003This article is dedicated to the 85th birthday of the discover of the sex-peptide, Prof. Dr. Pei Shen Chen, Zoological Institute, University of Zürich, Switzerland. P. S. Chen has served on the Editorial Board of Experientia (now CMLS) from 1974 to 1988.     

Cyclic CMP (cytidine 3′,5′-monophosphate) suppresses changes in human sperm amplitude of lateral head displacement and hyperactivation

Summary The dibutyryl analog of cCMP suppressed sperm amplitude of lateral head displacement and hyperactivation. Sperm motility was inhibited by dibutyryl cCMP with a shift toward less linear trajectory sperm movements. The results suggest a role of cCMP as an inhibitory signal on sperm motility patterns related to sperm capacitation.     

Cyclic CMP (cytidine 3',5'-monophosphate) suppresses changes in human sperm amplitude of lateral head displacement and hyperactivation

The dibutyryl analog of cCMP suppressed sperm amplitude of lateral head displacement and hyperactivation. Sperm motility was inhibited by dibutyryl cCMP with a shift toward less linear trajectory sperm movements. The results suggest a role of cCMP as an inhibitory signal on sperm motility patterns related to sperm capacitation.     

Hormonal facilitation in the release of sperm from the spermatheca of the red-spotted newt

Summary Several neurotransmitters and hormones with potential to trigger a simultaneous contraction of the oviducts and the spermathecal myoepithelium were examined. Saline (0.05 ml), or 0.05 ml saline plus acetylcholine (9 mg), norepinephrine (50 g), arginine-vasotocin (25 units), prostaglandin F₂(3g) were injected into the spermathecal region of female newts (n=24 per group). The numbers of sperm present in the cloacae of prostaglandin-injected animals (107±30 SEM) were significantly greater than the numbers detected in saline (27±5 SEM) and in uninjected (14±3 SEM) controls. Smaller and less consistent increases in the numbers of sperm were detected in the vasotocin- and norepinephrine-injected groups. Study of sections from ovulating female newts failed to produce evidence that pressure from the passage of ova through the posterior portion of the oviduct forced sperm from the spermatheca. Observations indicate an active role for the spermathecal myoepithelium in the discharge of stored sperm and of a role for prostaglandin F₂ in triggering that discharge.     

Association of Golgi vesicles containing acid phosphatase with the chromatoid body of rat spermatids

The relationship between the Golgi apparatus and the small vesicles associated with the chromatoid body was investigated using a cytochemical technique. It was observed that in early spermatids, when the chromatoid body appears in close contact with the Golgi complex, and all through its migration to the caudal pole of the nucleus, some of the vesicles that accompany the organelle display acid phosphatase activity. It is concluded that these smooth vesicles originate in the Golgi apparatus.     

Free amino acid composition of the intestinal contents,intestinal cells and hemolymph ofPhilosamia cynthia larvae

Summary Free amino acid composition of the intestinal contents, intestinal cells and hemolymph has been determined in larvae of the mothPhilosamia cynthia. From the hemolymph/lumen concentration ratio, an active transport could be inferred for neutral and basic amino acids. The values of cell/lumen and hemolymph/cell ratios suggested that the active step in the transport mechanism could be localized at the luminal pole of the enterocyte for neutral amino acids (except aromatic amino acids) and at the basolateral pole of the enterocyte for basic amino acids (except arginine).This work was supported by grants from Italian Consiglio Nazionale delle Ricerche and from Ministero della Pubblica Istruzione, Rome. The authors are indebted to Prof. V. Capraro for helpful discussion.     

Observations on the dynamics of argyrophilic nucleolar material in the nuclei of mice spermatids

Applying a new silver staining technique it could be shown that in very early spermatids strong argyrophilia in nucleoli is confined to their granular and fibrillar components; fibrillar centers are devoid of silver. During subsequent developmental stages remnants of these nucleolar components are present in the form of intensively silver stained clusters of coiled fibers. As chromatin condensation proceeds, these fibrous structures decrease in size and density and are finally completely absent. The nucleus of the mature sperm contains only the space in which they formerly existed, now silver negative, as the so-called 'nuclear vacuole'.     

Sperm predence in a hermaphroditic nematode (Caenorhabditis elegans) is due to competitive superiority of male sperm

When male and hermaphroditeCaenorhabditis elegans mate, the male's sperm outcompete the hermaphrodite's own sperm and fertilize a majority of the offspring. Here, we investigate the mechanism of male sperm precedence. We rule out the possibility that male sperm are stronger and more competitive because they are activated later than hermaphrodite sperm. We also find that a previously known gender difference in sperm activation does not influence sperm competition. Male sperm, rinsed free of seminal fluid, retained the capacity to take precedence after artificial insemination. Therefore, we conclude that male sperm themselves are competitively superior to hermaphrodite sperm. This trait maximizes outcrossing after mating and may increase both genetic diversity and heterozygosity of offspring whose parents, due to self-fertilization, may be highly homozygous.     

Nuclear blebs in oocytes of the fishClarias batrachus

Summary Large nuclear blebs have been observed in oocytes of the fish,Clarias batrachus. The bleb, which contains nucleoplasm, is finally extruded from the nucleus. The extruded body is DNase-sensitive, and it implies that some amount of DNA is extruded from the nucleus of these oocytes.Acknowledgments. We are thankful to Dr. R.K. Sharan, Department of Zoology, Patna University, for providing laboratory facilities, and to Dr V.G. Jhingran and Dr P.V. Dehadrai of ICAR for their keen interest in the work.