Cryopreservation of sperm from the coral Acropora humilis

Corals are sensitive to minute changes in their environments, and their continued existence is substantially threatened by the increasing number of destructive anthropogenic activities and unprecedented rates of climate change. Although cryopreservation has been successfully to preserve mammalian gametes for decades, coral cryopreservation was attempted for the first time less than 15 years ago, and freezing protocols exist for only a handful of coral species. The present study developed a cryopreservation protocol for the sperm of the common Indo-Pacific reef-builder Acropora humilis. Colonies of reefs of Sattahip Bay, Chonburi Province, Thailand were collected from 3 m depth with a mesh net during a spawning event. Immediately after collection, the sperm were isolated and subjected to a two-step freezing method featuring DMSO, polyethylene glycol, or methanol as the cryoprotectant. Viability and motility were assessed via a bioluminescence technique and a “computer-assisted semen analysis, and it was found that a 15-min equilibration with 2 M DMSO followed by cooling at 41.7 °C was the optimum cryopreservation protocol for A. humilis sperm. The post-thaw sperm achieved 45% fertilization success, and 35% of the fertilized eggs developed into blastopore larvae. The present optimized protocol can therefore facilitate the preservation of sperm for future propagation efforts of this species and provide an experimental platform for optimizing cryopreservation protocols for gametes of other scleractinian coral species.     

Rat Sertoli and spermatogenic cells express a similar gene, and its product is antigenically related to an outer dense fiber-associated protein.

We have previously reported that a heterodimeric protein secreted by rat Sertoli cells is antigenically related to a protein associated with outer dense fibers of the sperm tail. Therefore, we have explored the possibility that Sertoli and spermatogenic cells express a similar gene encoding a homologous protein. A Sertoli cell heterodimeric protein cDNA probe recognizes specific mRNA in pachytene and round spermatids fractionated by centrifugal elutriation; however, this specific mRNA was less prominent than in cultured Sertoli cells. In agreement with these observations, in situ hybridization experiments show that Sertoli cells are predominantly engaged in active heterodimeric protein mRNA synthesis, while meiotic prophase spermatocytes and spermatids also show significant but less abundant specific mRNA. Immunoblotting experiments demonstrate that, while Sertoli cells synthesize a heterodimeric protein consisting of two disulfide-linked components with molecular masses of 45 and 35 kD, both primary spermatocytes and round spermatids synthesize single 30 kD monomers not associated by disulfide linkage but recognized by antisera to Sertoli cell heterodimeric protein. Immunoblotting and immunogold electron microscopic studies show that antisera to Sertoli cell heterodimeric protein recognize a protein associated with outer dense fibers. This immunoreactivity was abolished by a 5-min pronase treatment, without affecting the integrity of outer dense fibers. Results of this study and previous studies demonstrate that both Sertoli and spermatogenic cells express a similar gene and that an antigenically related product encoded by this gene becomes associated with outer dense fibers during their assembly at spermiogenesis.     

Sperm cell surface characteristics of Plumbago zeylanica L. in relation to transport in the embryo sac

Myosin associated with the male germ cells of angiosperms interacts with actin, promoting transport of the non-motile generative and later sperm cells in the pollen tube. Myosin localizing on the sperm cell plasma membrane seems negligible in Plumbago, as reflected by the absence of: (i) anti-myosin labeling using immunoelectron microscopy, (ii) sperm motility on actin matrices, and (iii) electrophoretic movement changes after addition of antibody. Sperm cells injected directly into actively streaming Nitella internodal cells, however, follow actin bundles and their movement is sensitive to ATP and Mg²⁺. This may be based on simple charge binding since negatively charged latex beads also migrate on actin, whereas neutral or positively-charged latex beads do not. Sperm cells are negatively charged according to capillary microelectrophoresis, whereas killed sperm cells, which are positively charged do not migrate. The sperm cell that normally fertilizes the egg has a higher calculated charge (8.277 × 10³ esu/cm²) compared with the sperm cell that fuses with the central cell (6.120 × 10³ esu/cm²). Received: 15 December 1998 / Accepted: 21 January 1999     

Petrosal bones of metatherian mammals from the Late Palaeocene of Itaboraí (Brazil), and a cladistic analysis of petrosal features in metatherians

Metatherian petrosal bones were recovered from the early Late Palaeocene Itaboraí, Brazil, and are formally described. A cladistic analysis of the distribution of 56 petrosal and basicranial characters among extant and fossil metatherians was conducted, resulting in seven parsimonious trees. Relationships among metatherian ingroup taxa are congruent with current understanding of metatherian phylogeny. Metatheria is diagnosed by eight petrosal synapomorphies: stapedial artery absent in adults; reduced, intramural prootic canal; extrabullar internal carotid artery; inferior petrosal sinus between petrosal, basisphenoid, and basioccipital; cava supracochleare and epiptericum completely separated; reduction of the lateral flange; reduction of the anterior lamina; separation of the jugular foramen from the opening for the inferior petrosal sinus. The Palaeocene taxa Mayulestes , Pucadelphy s, and Andinodelphys from Tiupampa, and Petrosal Type II from Itaboraí are the sister groups of all other South American and Australian metatherians. This analysis confirms previous results showing the South American 'monito del monte' Dromiciops nested within the Australasian radiation. Within this australidelphian clade, Dromiciops is closely related to the dasyurids. The South American Caenolestes appears more closely related to the Australidelphia than to the South American didelphids. The Petrosal Types I, III, IV and V from Itaboraí are the stem taxa of the clade Australidelphia plus Caenolestes . The significant synapomorphies supporting this relationship are: enlargement of the fossa subarcuata that produces a bulbous ventral aspect of the mastoid and loss of post-temporal canal.  Journal compilation © 2007 The Linnean Society of London, Zoological Journal of the Linnean Society , 2007, 150 , 85–115. No claim to original French government works.     

Morphometric analysis of interspecific and microgeographic variation of crayfish from a Mexican cave

Two undescribed species of Procambarus crayfish (Crustacea, Decapoda) inhabit the subterranean stream running in the recently explored Cueva de Los Camarones, Chiapas, Mexico. Individuals ranging from dark, robust, large-eyed forms with an epigean mien to pale, comparatively slender, microphthalmic ones resembling related troglobites were found, but only the extremes could be distinguished visually. Evolutionary relationships among individuals were investigated morphometrically by means of multivariate analysis'on 14 body dimensions. These analyses revealed a significant discontinuity between the two species, mainly determined by the shape of the rostrum, chelae and telson. The occurrence of a few aberrant phenotypes may indicate the existence of some gene flow between the two species.
Microgeographic variation was demonstrated within the less cavernicolous species. The characters involved were the same as those that discriminate between the two species. These results are compared with allozymic data presented in a parallel paper and the role of genie introgression and environmental induction are discussed.     

Geographic variation in jungle cat (Felis chaus Schreber, 1777) (Mammalia,Carnivora, Felidae) body size: is competition responsible?

There is a striking difference in body size of jungle cats ( Felis chaus ) in the west and the east of their distribution, with Israeli cats being 43% heavier than Indian cats. We tested the hypothesis that increasing competition from other small felids towards the east is responsible for the difference in body size. We measured jungle cat skulls for eight cranial and dental variables and related these to independent variables such as species richness (local and regional), latitude, longitude, temperature, and precipitation. Data from a narrow band between latitudes 24.0°N and 33.9°N, where Bergmann's rule was largely not observed, showed that the western population (≤ 50.0°E longitude) of jungle cats is larger than the eastern (> 60.0°E longitude) population with the size difference being most evident in the upper carnassials (P⁴L). Species richness at the regional level showed a significant negative relation to P⁴L. An even spacing in condylobasal length for a small-cat guild from India through null model analysis indicated the occurrence of character displacement. The results support the hypothesis that competition is responsible for geographical variation in jungle cat body size in the region where Bergmann's rule does not apply. © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 163–172.     

三尖杉属受精作用的研究

三尖杉属的精原细胞有一类似银杏生毛体的星状体结构,它分裂产生的2个精子,在大小与形态上都基本相同,而且在精子细胞质中具有拟核仁颗粒存在。上述结构是三尖杉属的重要特点之一。本属植物的成熟卵细胞特别长,细胞质中有丰富的拟核仁结构,卵核下方具2—3团浓稠的细胞质团,这些结构很像穗花杉的卵细胞。三尖杉属的受精作用,属于有丝分裂后类型,这种类型只在松科和三尖杉科中发现。受精后,卵细胞发生强烈的极性分化,上部细胞质变成高度液泡化;相反,下部细胞质则聚集大量蛋白泡和拟核仁颗粒。     

A multivariate comparison between three NW. European populations ofArtemisia norvegica (Asteraceae) by means of chemometric and morphometric data

Specimens from Scotland, S. and C. Norway were grown in the botanical garden of Bergen, Norway. Some of the Scottish specimens came from a meristem tissue culture. The specimens were compared by a principal component analysis of lipids and related compounds, and of morphological characters from leaves and flowers. The populations differed from each other, but some overlap was found in leaf characters. The results are discussed in relation to distribution and immigration history, and it is argued that the differences among the populations may have evolved in postglacial time.     

The spatial association of the sperm cells and vegetative nucleus in the pollen grain ofBrassica

Summary In mature viable pollen ofBrassica oleracea, the pair of sperm cells and the nucleus of the vegetative cell are linked to form a structured unit we term the male germ unit. The sperm cells are held within a common periplasm and have no cell walls. Each sperm cell has a central globular body containing the nucleus surrounded by several evaginations which provide the means for linkage between them. One sperm cell, usually that closest to the nucleus of the vegetative cell contains most of mitochondria profiles (plastids are absent). This sperm cell appears to be linked by its protoplasmic evaginations to the envelope of the vegetative nucleus. The role of this unit in interactions with the female gametic complex is considered.