Chromosomal rearrangements and the first indication of an ♀X1X1X2X2/♂X1X2Y sex chromosome system in Rineloricaria fishes (Teleostei: Siluriformes)

Rineloricaria is the most diverse genus within the freshwater fish subfamily Loricariinae, and it is widely distributed in the Neotropical region. Despite limited cytogenetic data, records from southern and south-eastern Brazil suggest a high rate of chromosomal rearrangements in this genus, mirrored in remarkable inter- and intraspecific karyotype variability. In the present work, we investigated the karyotype features of Rineloricaria teffeana, an endemic representative from northern Brazil, using both conventional and molecular cytogenetic techniques. We revealed different diploid chromosome numbers (2n) between sexes (33♂/34♀), which suggests the presence of an ♀X₁X₁X₂X₂/♂X₁X₂Y multiple sex chromosome system. The male-limited Y chromosome was the largest and the only biarmed element in the karyotype, implying Y-autosome fusion as the most probable mechanism behind its origination. C-banding revealed low amounts of constitutive heterochromatin, mostly confined to the (peri)centromeric regions of most chromosomes (including the X₂ and the Y) but also occupying the distal regions of a few chromosomal pairs. The chromosomal localization of the 18S ribosomal DNA (rDNA) clusters revealed a single site on chromosome pair 4, which was adjacent to the 5S rDNA cluster. Additional 5S rDNA loci were present on the autosome pair 8, X₁ chromosome, and in the presumed fusion point on the Y chromosome. The probe for telomeric repeat motif (TTAGGG)_n revealed signals of variable intensities at the ends of all chromosomes except for the Y chromosome, where no detectable signals were evidenced. Male-to-female comparative genomic hybridization revealed no sex-specific or sex-biased repetitive DNA accumulations, suggesting a presumably low level of neo-Y chromosome differentiation. We provide evidence that rDNA sites might have played a role in the formation of this putative multiple sex chromosome system and that chromosome fusions originate through different mechanisms among different Rineloricaria species.     

Synchrony,leadership, and association in male Indo-pacific bottlenose dolphins (Tursiops aduncus)

Male Indo-pacific bottlenose dolphins in Shark Bay, Western Australia, have converged with humans in the formation of nested male alliances and the use of synchrony in alliance behavior. Further, the strength of association among allied male dolphins varies and the stability of alliances correlates with the rate that males consort with estrus females (and is thus a possible indicator of dominance). To examine the possibility that synchrony reflects alliance association strength and dominance relationships, we analyzed videotapes from focal follows of two groups of males that reflect the range of alliance size and the strength of association between individuals in the population. We examined two variables: leadership during synchronous behaviors, based on which animal in a synchronously surfacing pair surfaced first, and the degree of synchrony, based on temporal differences in synchronous surfacing. We predicted that closer associates would exhibit a greater degree of synchrony and that one dolphin in a dyad would consistently lead. Contrary to our predictions, the degree of synchrony was inversely related to strength of association within alliances. This surprising result suggests that individuals with less secure bonds may strive more to achieve synchrony. We found no evidence of leadership during synchronous surfacing or between synchrony and other behavioral variables. Proximate mechanisms for synchronous behavior, such as entrainment and mutual motor imitation (“the mirror game” paradigm), may inhibit leadership in this context. Our results show that synchrony during surfacing is not a useful behavior to examine for dominance relationships in wild dolphins but it may be a useful tool to examine variation in alliance relationships.     

L-Arginine restores splenocyte functions after trauma and hemorrhage potentially by improving splenic blood flow

Several studiesindicate that immune responses are markedly depressed early after onsetof hemorrhage. Decreased organ blood flow has been implicated in thepathophysiology of altered immune responses after trauma-hemorrhage. Inthis regard, administration ofL-arginine has been shown torestore depressed intestinal and hepatic blood flow aftertrauma-hemorrhage, probably due to provision of substrate forconstitutive nitric oxide synthase (cNOS). It remains unknown, however,whether administration ofL-arginine also amelioratesdepressed splenic blood flow and whether this agent has any salutaryeffects on depressed splenocyte functions after trauma-hemorrhage. Malerats underwent sham operation or laparotomy and were bled to andmaintained at a mean arterial blood pressure of 40 mmHg until 40% ofmaximum shed blood volume (MBV) was returned as Ringer lactate (RL).Hemorrhaged rats were then resuscitated with RL (4 times MBV over 1 h).During resuscitation, rats received 300 mg/kgL-arginine or saline (vehicle)intravenously; 4 h later, splenic blood flow, splenocyte proliferation,and splenocyte interleukin (IL)-2 and IL-3 were determined.Administration of L-arginineimproved depressed splenic blood flow and restored depressed splenocytefunctions after trauma-hemorrhage. Therefore, provision ofL-arginine during resuscitationafter trauma-hemorrhage should be considered a novel and safe approachfor improving splenic organ blood flow and depressed splenocytefunctions under such conditions.

     

Tissue distribution of a plasmid DNA encoding Hsp65 gene is dependent on the dose administered through intramuscular delivery

In order to assess a new strategy of DNA vaccine for a more complete understanding of its action in immune response, it is important to determine the in vivo biodistribution fate and antigen expression. In previous studies, our group focused on the prophylactic and therapeutic use of a plasmid DNA encoding the Mycobacterium leprae 65-kDa heat shock protein (Hsp65) and achieved an efficient immune response induction as well as protection against virulent M. tuberculosis challenge. In the present study, we examined in vivo tissue distribution of naked DNA-Hsp65 vaccine, the Hsp65 message, genome integration and methylation status of plasmid DNA. The DNA-Hsp65 was detectable in several tissue types, indicating that DNA-Hsp65 disseminates widely throughout the body. The biodistribution was dose-dependent. In contrast, RT-PCR detected the Hsp65 message for at least 15 days in muscle or liver tissue from immunized mice. We also analyzed the methylation status and integration of the injected plasmid DNA into the host cellular genome. The bacterial methylation pattern persisted for at least 6 months, indicating that the plasmid DNA-Hsp65 does not replicate in mammalian tissue, and Southern blot analysis showed that plasmid DNA was not integrated. These results have important implications for the use of DNA-Hsp65 vaccine in a clinical setting and open new perspectives for DNA vaccines and new considerations about the inoculation site and delivery system.     

Improved synthesis of the sodium salt of dl-α,β-dihydroxyisovaleric acid

A one-step synthesis of sodium dl-α,β-dihydroxyisovalerate involving the hydroxylation of 3,3-dimethylacrylic acid is described. This procedure is easier and more economical than those published previously. The material synthesized is shown to be a substrate for Salmonella typhimurium dihydroxyacid dehydratase (the product of the ilvD gene).     

Adult male Cuvier's beaked whales (Ziphius cavirostris) engage in prolonged bouts of synchronous diving

Studies of the social behavior of Cuvier's beaked whales (Ziphius cavirostris) are challenging because of their deep-water habitat usually far from shore and the limited time they spend at the surface. The sociality of these deepest diving mammals is of interest, however, especially for our understanding of how social systems evolve in extreme habitats. High levels of scarring suggest that males compete agonistically for access to females and so we predicted that associations among adult males would be unstable due to competitive exclusion. We tested this prediction by evaluating the diving behavior of animals within social groups off Cape Hatteras, North Carolina, considering diving synchrony a proxy for group membership. Using data from satellite-linked depth-recording tags, we found that adult male–male pairs showed extended periods of synchrony in diving behavior, while all pairs that included an adult male with an individual of another age and/or sex dove synchronously for less than a day. We assessed three hypotheses to explain these surprising results: sexual segregation; extended bouts of male–male competition; and the presence of male alliances. Finally, we considered testable predictions to distinguish among these explanations.     

Molecular mechanisms of regulation of neuronal muscarinic receptor sensitivity

Like other neurotransmitter receptors, muscarinic acetylcholine receptors are subject to regulation by the state of receptor activation. Prolonged increases in the concentration of muscarinic agonists result in a decrease in receptor density and loss of receptor sensitivity, both in vivo and in vitro. On the other hand, when the receptor is deprived of acetylcholine for a long duration in vivo, the receptor becomes more sensitive in responding to muscarinic agonists. However, it has been more difficult to demonstrate increases in receptor concentration that accompany this supersensitive state. The purpose of this review is to provide current information related to the characteristics of muscarinic receptor regulation and the molecular mechanisms underlying this phenomenon, regarding both the density of receptors and their transduction mechanisms. Furthermore, possible feedback regulatory roles of different second messenger signals are discussed. Particular emphasis is dedicated to molecular mechanisms of regulation of neuronal muscarinic receptors.