On spermiogenesis, sperm cell morphology and accompanying secretions of Copidognathus (Acari: Halacaridae)

The genus Copidognathus includes one-third of the species of Halacaridae described to date. This article describes spermiogenesis, sperm cell morphology and accompanying secretions from three species of Copidognathus. Initial spermatids have electron-dense cytoplasm with scattered mitochondria, a well-developed Golgi body, and nuclei with patches of heterochromatin. The cytoplasm and nuclei of these cells undergo intense swelling. The second spermatids are large electron-translucent cells, with small mitochondria in row along the remains of the endoplasmatic reticulum. In the succeeding stage, most of the cytoplasmatic structures and mitochondria have disappeared or have undergone profound transformations. Nuclei and cells elongate and chromatin begins to condense near the nuclear envelope. An acrosomal complex appears at the tip of the nucleus. The acrosomal filament is thick and runs the entire length of the nucleus. Plasmalemmal invaginations at the cell surface give rise to tubules filled with an electron-dense material. Sperm cell maturation is completed in the ventral portion of the germinal part, near the testicular lumen. As a final step in spermiogenesis, cytoplasm of the last spermatid undergoes a moderate condensation and the cariotheca disappears. Mature sperm cells were found in a matrix of ""simple"" and ""complex"" corpuscles, the latter consisting of flattened, spindle-shaped secreted bodies. Rather than in individual sperm aggregates, spermatozoa were contained in a single droplet inside the vas deferens, on a large secretion mass, structured as rows of platelets sunk in a fine grained matrix. Each mature sperm cell is covered by a thick secreted coat. In contrast to the genera Rhombognathus and other Actinotrichida, Copidognathus displays a set of features that must be regarded as apomorphic. The absence of usual mitochondria, the presence of electro-dense tubules and secretions similar to those present in Thalassarachna and Halacarellus, and the pattern of nuclear condensation are possibly shared apomorphies with these latter genera. (C) 2010 Elsevier GmbH. All rights reserved.

Production of Bovine Hand-Made Cloned Embryos by Zygote-Oocyte Cytoplasmic Hemi-complementation

The aim of this study was to evaluate the effect of the cytoplast type and activation process on development of cloned embryos. Bovine oocytes (MII) or zygotes at the one-cell stage (IVF) were manually bisected and segregated in MII or IVF hemi-cytoplasts or hemi-karyoplasts. Adult skin cells from a bovine female were used as nucleus donors (SC). Experimental groups were composed of IVF embryos; parthenogenetic embryos; handmade cloned (HMC) embryos; and reconstructed HMC embryos using IVF hemi-cytoplast + MII hemi-cytoplast + SC (G-I); IVF hemi-cytoplast + IVF hemi-cytoplast + SC (G-II); MII hemi-cytoplast + IVF hemi-karyoplast (G-III); and IVF hemi-cytoplast + IVF hemi-karyoplast (G-IV). Embryos from G-I to G-IV were allocated to subgroups as sperm-activated (SA) or were further chemically activated (SA + CA). Embryos from all groups and subgroups were in vitro cultured in the WOW system. Blastocyst development in subgroup G-I SA (28.2%) was similar to IVF (27.0%) and HMC (31.4%) controls, perhaps due to a to a more suitable activation process and/or better complementation of cytoplasmic reprogramming factors, with the other groups and subgroups having lower levels of development. No blastocyst development was observed when using IVF hemi-karyoplasts (G-III and G-IV), possibly due to the manipulation process during a sensitive biological period. In summary, the presence of cytoplasmic factors from MII hemi-oocytes and the sperm activation process from hemi-zygotes appear to be necessary for adequate in vitro development, as only the zygote-oocyte hemi-complementation was as efficient as controls for the generation of bovine cloned blastocysts.

Testicular traits as selection criteria for young Nellore bulls

The objective of this study was to evaluate the possible use of biometric testicular traits as selection criteria for young Nellore bulls using Bayesian inference to estimate heritability coefficients and genetic correlations. Multitrait analysis was performed including 17,211 records of scrotal circumference obtained during andrological assessment (SCAND) and 15,313 records of testicular volume and shape. In addition, 50,809 records of scrotal circumference at 18 mo (SC18), used as an anchor trait, were analyzed. The (co) variance components and breeding values were estimated by Gibbs sampling using the Gibbs2F90 program under an animal model that included contemporary groups as fixed effects, age of the animal as a linear covariate, and direct additive genetic effects as random effects. Heritabilities of 0.42, 0.43, 0.31, 0.20, 0.04, 0.16, 0.15, and 0.10 were obtained for SC18, SCAND, testicular volume, testicular shape, minor defects, major defects, total defects, and satisfactory andrological evaluation, respectively. The genetic correlations between SC18 and the other traits were 0.84 (SCAND), 0.75 (testicular shape), 0.44 (testicular volume), -0.23 (minor defects), -0.16 (major defects), -0.24 (total defects), and 0.56 (satisfactory andrological evaluation). Genetic correlations of 0.94 and 0.52 were obtained between SCAND and testicular volume and shape, respectively, and of 0.52 between testicular volume and testicular shape. In addition to favorable genetic parameter estimates, SC18 was found to be the most advantageous testicular trait due to its easy measurement before andrological assessment of the animals, even though the utilization of biometric testicular traits as selection criteria was also found to be possible. In conclusion, SC18 and biometric testicular traits can be adopted as a selection criterion to improve the fertility of young Nellore bulls.

Chronic colitis associated with HIV infection can be related to intraepithelial infiltration of the colon by CD8+T lymphocytes

Gastrointestinal complications in AIDS patients with diarrhoea are common clinical manifestations, frequently diagnosed by colonoscopy as non-specific colitis. We retrospectively study colon biopsies diagnosed as chronic colitis associated with HIV (CCH). Biopsies were sorted as patients with AIDS (serum CD4 < 200 cell/mm(3)) but without any clear infectious process (n = 12) and patients without HIV infection (n = 24). There are low numbers of CD4+ T lymphocytes in lamina propria of AIDS patients, but CD8+ T populations in this area appear to be similar in all studied groups, regardless of HIV infection or laboratory evidence of a specific agent. We found the clear evidence of CD8+ T cells infiltration in colonic mucosa in HIV patients with microscopic colitis. An imbalance of lymphocyte subpopulations in the colon, both in the lamina propria and epithelium, could result in an intraepithelial CD8 infiltration, involved in the pathogenesis of CCH in AIDS patients.

Alternative mating tactics in dimorphic males of the harvestman Longiperna concolor (Arachnida: Opiliones)

Intense male-male competition for females may drive the evolution of male morphological dimorphism, which is frequently associated with alternative mating tactics. Using modern techniques for the detection of discontinuous allometries, we describe male dimorphism in the Neotropical harvestman Longiperna concolor, the males of which use their elongated, sexually dimorphic legs IV in fights for the possession of territories where females lay eggs. We also tested three predictions related to the existence of alternative mating tactics: (1) if individuals with relatively longer legs IV (majors) are more likely to monopolize access to reproductive resources, they are expected to remain close to stable groups of females more than individuals with relatively shorter legs IV (minors) do; (2) if minors achieve fertilization by moving between territories, they are expected to be less faithful to specific sites; and (3) majors should be observed in aggressive interactions more often. We individually marked all the individuals from a population of Longiperna during the reproductive season and recorded the location of each sighting for males and females as well as the identity of males involved in fights. Majors were more likely to have harems, and large majors were even more likely to do so. Majors were more philopatric and all males involved in fights belonged to this morph. These results strongly suggest that the mating tactic of the majors is based on resource defense whereas that of the minors probably relies on sneaking into the territories of the majors and furtively copulating with females.

Reproductive cycle of Ophionereis reticulata (Ophiuroidea, Echinodermata) on the southeast coast of Brazil

The reproductive cycle of Ophionereis reticulata, a common sediment-rocky shore-interface ophiuroid, was examined monthly from January 2002 to January 2003 at Praia Grande beach (Sao Sesbatiao, state of Sao Paulo, Brazil). Mature individuals were found from January 2002 to April 2002 and from November 2002 to January 2003. Spawning was regcorded from January 2002 to March 2002 and from November 2002 to January 2003. Mature sperm is still present in April which could be involved in the fertilization of the last oocytes spawned in March and April. November and December marked the final point in the maturation process, with a high concentration of yolk and lipid nutrients in the oocytes. Ophionereis reticulata showed, for the period analyzed, a single spawning period, during spring and summer.

Mating behavior of Sickius longibulbi (Araneae, Theraphosidae, Ischnocolinae), a spider that lacks spermathecae

We describe the mating behavior in the spermatheca-lacking theraphosid species Sickius longibulbi Soares & Camargo 1948. The behavior in captivity of nine pairs of S. longibulbi was videotaped and analyzed. The matting of this species presented an uncommon theraphosid pattern. There is little in the way of overt courtship by the male, the primary behavior seen being the male`s use of legs I and II to touch the female`s first pairs of legs and her chelicerae. Sometimes the male clasped one of the female`s first pairs of legs, bringing her close to him. While the female raised her body, the male clasped her fangs and held her tightly with his legs III wrapped around her prosoma. The male seemed to try to knock the female down, pushing her entire body until she lay on her dorsum. In this phase we observed the male biting the female on the sternum or on the leg joints. When the female fell, the male attempted to position himself at an angle of 90 degrees from the female. These movements appear to demand a lot of energy, particularly because the female is not passive during the mating. Our findings suggest that copulating in this position is, for the male, more successful than adopting other positions because it allows his extremely long palpal bulbs to deposit more sperm in the female oviduct where - since she lacks spermathecae - she retains the sperm. We suggest that the further he reaches into the oviduct, the greater the chance that he will fertilize the female`s eggs.

Immunohistochemical analysis of neuron types in the mouse small intestine

The definition of the nerve cell types of the myenteric plexus of the mouse small intestine has become important, as more researchers turn to the use of mice with genetic mutations to analyze roles of specific genes and their products in enteric nervous system function and to investigate animal models of disease. We have used a suite of antibodies to define neurons by their shapes, sizes, and neurochemistry in the myenteric plexus. Anti-Hu antibodies were used to reveal all nerve cells, and the major subpopulations were defined in relation to the Hu-positive neurons. Morphological Type II neurons, revealed by anti-neurofilament and anti-calcitonin gene-related peptide antibodies, represented 26% of neurons. The axons of the Type II neurons projected through the circular muscle and submucosa to the mucosa. The cell bodies were immunoreactive for choline acetyltransferase (ChAT), and their terminals were immunoreactive for vesicular acetylcholine transporter (VAChT). Nitric oxide synthase (NOS) occurred in 29% of nerve cells. Most were also immunoreactive for vasoactive intestinal peptide, but they were not tachykinin (TK)-immunoreactive, and only 10% were ChAT-immunoreactive. Numerous NOS terminals occurred in the circular muscle. We deduced that 90% of NOS neurons were inhibitory motor neurons to the muscle (26% of all neurons) and 10% (3% of all neurons) were interneurons. Calretinin immunoreactivity was found in a high proportion of neurons (52%). Many of these had TK immunoreactivity. Small calretinin neurons were identified as excitatory neurons to the longitudinal muscle (about 20% of neurons, with ChAT/calretinin/+/- TK chemical coding). Excitatory neurons to the circular muscle (about 10% of neurons) had the same coding. Calretinin immunoreactivity also occurred in a proportion of Type II neurons. Thus, over 90% of neurons in the myenteric plexus of the mouse small intestine can be currently identified by their neurochemistry and shape.

CCR5 Mediates Pro-osteoclastic and Osteoclastogenic Leukocyte Chemoattraction

Periodontal disease (PD) progression involves the selective leukocyte infiltration into periodontium, supposedly mediated by the chemokine/chemokine receptor system. In this study, we investigated the role of chemokine receptor CCR5 in the immunoregulation of experimental PD in C57BL/6 (WT) and CCR5KO mice. Aggregatibacter actinomycetem comitans infection triggered the chemoattraction of distinct CCR5+ leukocyte subpopulations (determined by flow cytometry): CCR5+F4/80+ leukocytes, which co-express CD14, CCR2, TNF-alpha, and IL-1 beta, indicative of activated macrophages; and CCR5+CD4+ cells, which co-express CXCR3, IFN-gamma, and RANKL, indicative of Th1 lymphocytes, therefore comprising pro-osteoclastic and osteoclastogenic cell subsets, respectively. CCR5KO mice presented a lower PD severity (lower inflammation and alveolar bone loss) when compared with the WT strain, since the migration of F4/80+, TNF-alpha+, CD4+, and RANKL+ cells specifically decreased due to the lack of CCR5. Also, ELISA analysis demonstrated that the production of TNF-alpha, IL-1 beta, IL-6, IFN-gamma, and RANKL in periodontal tissues was significantly decreased in the CCR5KO strain. The periodontal bacterial load and antimicrobial patterns were unaltered in CCR5KO mice. Our results demonstrate that the chemokine receptor is involved in the migration of distinct leukocyte subpopulations throughout experimental PD, being a potential target for therapeutic intervention in PD.

The Estrous Cycle Modulates Small Leucine-Rich Proteoglycans Expression in Mouse Uterine Tissues

In the pregnant mouse uterus, small leucine-rich proteoglycans (SLRPs) are drastically remodeled within a few hours after fertilization, suggesting that ovarian hormone levels modulate their synthesis and degradation. In this study, we followed by immunoperoxidase approach, the presence of four members of the SLRP family (decorin, lumican, biglycan, and fibromodulin) in the uterine tissues along the estrous cycle of the mouse. All molecules except fibromodulin, which predominates in the myometrium, showed a striking modulation in their distribution in the endometrial stroma, following the rise in the level of estrogen. Moreover, notable differences in the distribution of SLRPs were observed between superficial and deep stroma, as well as between the internal and external layers of the myometrium. Only biglycan and fibromodulin were expressed in the luminal and glandular epithelia. All four SLRPs were found in cytoplasmic granules of mononucleated cells. The pattern of distribution of the immunoreaction for these molecules in the uterine tissues was found to be estrous cycle-stage dependent, suggesting that these molecules undergo ovarian hormonal control and probably participate in the preparation of the uterus for decidualization and embryo implantation. In addition, this and previous results from our laboratory suggest the existence of two subpopulations of endometrial fibroblasts that may be related to the centrifugal development of the decidua. Anat Rec, 292:138-153, 2009. (c) 2008 Wiley-Liss, Inc.

Differential expression of new LTA splice variants upon lymphocyte activation

Lymphotoxin alpha (LTA) is a member of the TNF cytokine superfamily, produced principally by lymphocytes. It plays an important role in immune and inflammatory responses. Many TNF superfamily members have functionally important isoforms generated by alternative splicing but alternative splicing of LTA has never been studied. The known LTA protein is encoded by a transcript containing four exons. Here we report seven new LTA splice variants, three of them evolutionary conserved. We demonstrate their presence in cytoplasmic RNA suggesting that they could be translated into new LTA isoforms. We observed that their expression is differentially regulated upon activation of peripheral blood mononuclear cells and lymphocyte subpopulations (CD4+, CD8+, and CD19+). Our data suggest that the new LTA splice variants might play a role in the regulation of the immune response. (C) 2007 Elsevier Ltd. All rights reserved.

Potassium Current in Mature Bovine Spermatozoa

Ion channels have been assigned a pivotal importance in various sperm functions and are therefore promising targets for contraceptive development. The lack of data on channel functionality and pharmacology has hampered this goal. This is a consequence of technical problems of applying electrophysiological techniques to spermatozoa due to their small size and form. By using a laminin coating to increase adherence of spermatozoa and nystatin in the patch pipette for pore formation, we have adapted the whole-cell recording technique to study currents in mature uncapacitated bovine spermatozoa. Employing these conditions, in the head region, patched spermatozoa could be transferred into the whole-cell configuration. For the first time we document an outward rectifying current in mature bovine spermatozoa was blocked by tetraethyl ammonium (TEA) chloride. The observation of a shift in the reversal potential as a response to changes in the extracellular concentration of K+ ions allowed us to identify this current as K+ selective. This result shows that K+ channels in the head region of mature uncapacitated bovine spermatozoa can be suitably investigated using the whole-cell recording patch-clamp technique.