Linking Acrosome Size and Genetic Divergence in an Inter-Oceanic Mussel from the Pacific and Atlantic Coasts: A Case of Incipient Speciation?

In recent years, advances in analyses of the sperm morphology and genetics of Perumytilus purpuratus have allowed to two evolutionary scenarios for this mussel to be suggested: (1) the scenario of cryptic species and (2) the scenario of incipient or in progress speciation. For a better understanding of the evolutionary history of P. purpuratus, we performed extensive sampling along a latitudinal gradient of ca. 7180 km of coastline-from the Southern Pacific Ocean to the Atlantic Ocean-and we delved deeper into the sperm morphology of P. purpuratus, exploring its association with the phylogeny and population genetics to determine whether the variability in sperm traits between the northern and southern regions was a signal of cryptic or incipient species. Overall, our results showed that sperm sizes were strongly correlated with the genetic structure in males of P. purpuratus. We identified at 37° S on the Pacific coast a coincident break of both sperm size and genetic disruption that can be explained by historical events and postglacial recolonization as causal phenomena for the observed divergences. Furthermore, evidence of genetic admixture between lineages was found at 38° S, suggesting the presence of an introgressive hybridization zone and incomplete reproductive isolation in an in fraganti or incipient speciation process.

Metalloprotease ADAMTS-1 decreases cell migration and invasion modulating the spatiotemporal dynamics of Cdc42 activity.

ADAMTSs (A Disintegrin And Metalloproteinase with ThromboSpondin motifs) are secreted proteases dependent on Zn2+/Ca2+, involved in physiological and pathological processes and are part of the extracellular matrix (ECM). Here, we investigated if ADAMTS-1 is required for invasion and migration of cells and the possible mechanism involved. In order to test ADAMTS-1's role in ovarian cancer cells (CHO, NIH-OVCAR-3 and ES2) and NIH-3 T3 fibroblasts, we modified the levels of ADAMTS-1 and compared those to parental. Cells exposed to ADAMTS-1-enriched medium exhibited a decline in cell migration and invasion when compared to controls with or without a functional metalloproteinase domain. The opposite was observed in cells when ADAMTS-1 was deleted via the CRISPR/Cas9 approach. The decline in ADAMTS-1 levels enhanced the phosphorylated form of Src and FAK. We also evaluated the activities of cellular Rho GTPases from cell lysates using the GLISA® kit. The Cdc42-GTP signal was significantly increased in the CRISPR ADAMTS-1 ES-2 cells. By a Förster resonance energy transfer (FRET) biosensor for Cdc42 activity in ES-2 cells we demonstrated that Cdc42 activity was strongly polarized at the leading edge of migrating cells with ADAMTS-1 deletion, compared to the wild type cells. As conclusion, ADAMTS-1 inhibits proliferation, polarization and migration.

ADAMTS1 Supports Endothelial Plasticity of Glioblastoma Cells with Relevance for Glioma Progression.

Gliomas in general and the more advanced glioblastomas (GBM) in particular are the most usual tumors of the central nervous system with poor prognosis. GBM patients develop resistance to distinct therapies, in part due to the existence of tumor cell subpopulations with stem-like properties that participate in trans-differentiation events. Within the complex tumor microenvironment, the involvement of extracellular proteases remains poorly understood. The extracellular protease ADAMTS1 has already been reported to contribute to the plasticity of cancer cells. Accordingly, this basic knowledge and the current availability of massive sequencing data from human gliomas, reinforced the development of this work. We first performed an in silico study of ADAMTS1 and endothelial markers in human gliomas, providing the basis to further assess these molecules in several primary glioblastoma-initiating cells and established GBM cells with the ability to acquire an endothelial-like phenotype. Using a co-culture approach of endothelial and GBM cells, we noticed a relevant function of ADAMTS1 in GBM cells leading the organization of endothelial-like networks and, even more significantly, we found a blockade of the formation of tumor-spheres and a deficient response to hypoxia in the absence of ADAMTS1. Our data support a chief role of this protease modulating the phenotypic plasticity of GBM.

Notes on Mesocapromys sanfelipensis (Rodentia: Capromyidae) from Cuba.

The San Felipe Hutia, Mesocapromys sanfelipensis, is one of the most endangered species of rodents in the world, and little is known about its ecology, evolution, and ancient distribution. At present, this hutia has been found only in its type locality, Cayo Juan Garcia, a cay in the southwest Cuban insular platform. Here we report for the first time a well preserved fossil skull referred to this species, collected in Cueva del Indio, Mayabeque province, western Cuba. This specimen shows that the modern population of M. sanfelipensis is a marginal relic of its former distribution, a consequence of climatic, eustatic, and neotectonic changes in the last 8 ka years. Also, we reevaluate the cranial characters and measurements that correspond to M. sanfelipensis and found that two of the eight specimens referred to this species and deposited at the Instituto de Ecologia y Sistematica belong to Mesocapromys auritus. Finally, we include six unpublished photos of specimens of M. sanfelipensis captured in 1970 during two expeditions to Cayo Juan Garcia.

Tejido tiroideo y láser infrarrojo: un estudio morfométrico / Thyroid tissue and infrared laser: a morphometric study

La glándula tiroides posee gran importancia debido a la síntesis y secreción de hormonas, las cuales desempeñan funciones fundamentales para la mantención de la fisiología animal. En este contexto, el objetivo del presente trabajo consistió en determinar parámetros morfométricos de estructuras tiroideas sometidas a estimulaciones con láser infrarrojo (LIR). Para ello, 10 ratas Sprague Dawley de 3 meses de vida y peso aproximado de 200 g, fueron divididas en dos grupos de 5 animales cada uno: grupo control y grupo experimental. Estos últimos recibieron estimulaciones infrarrojas en la tiroides con dosis de 16 J/cm2 durante 15 días seguidos. Una vez sacrificadas las ratas, se extrajeron las glándulas tiroides las que fueron procesadas para microscopía óptica obteniéndose placas histológicas y micrografías con aumentos finales de hasta 1000 X. Se efectuaron estudios morfométricos para determinar en 40 placas, variaciones tisulares generadas por las inducciones infrarrojas, con especial énfasis en la disposición coloidal y dimensiones de folículos y células tiroideas. El análisis de las 40 placas histológicas generados por las inducciones del láser infrarrojo comparados con los controles, reveló que existen marcadas diferencias en todos los componentes del tejido tiroideo analizado, lo cual podría otorgar antecedentes de diferentes funcionalidades en el metabolismo de las glándulas.

The thyroid gland is of great importance because of the synthesis and secretion of hormones which play key roles in the maintenance of animal physiology. In this context, the aim of the present study was to determine morphometric parameters of thyroid structures subjected to infrared laser stimulation (ILS) and for this purpose, 10 Sprague Dawley rats, 3 months of age and weighing approximately 200 grams, were divided into two groups of 5 animals each: the control group and the remaining 5 animals constituting the experimental group received infrared stimulation in the thyroid with doses of 16 J/cm2 for 15 consecutive days. After the rats were sacrificed, the respective thyroids were removed and processed for optical microscopy. Histological plates and micrographs were obtained with final increases of up to 1000 X. Morphometric studies were carried out to determine the tissue variations generated by infrared inductions, with special emphasis on the colloidal arrangement and dimensions of follicles and thyroid cells. Our results revealed that there are marked differences in all the components of the analyzed thyroid tissue which could give antecedents of different functionalities in the metabolism of thyroid glands.

Efecto del láser infrarrojo sobre células tiroideas / Infrared laser effect on thyroid cells

Diez ratas Sprague Dawley de 4 meses de vida y peso aproximado de 250 g fueron divididas en dos grupos de 5 animales cada uno, el grupo A se mantuvo como control y los animales del grupo B recibieron estimulaciones con láser infrarrojo en la tiroides con dosis de 16 J/cm2 durante 15 días consecutivos. Posteriormente las ratas fueron sacrificadas, se extrajeron las respectivas tiroides siendo procesadas para microscopía óptica y se obtuvieron placas histológicas y micrografías de tiroides con aumentos finales de hasta 1000X, las cuales fueron sometidas a estudios morfométricos para determinar en 100 células foliculares: número, áreas y perímetro tanto celular como nuclear, además de disposición coloidal y presencia de vasos sanguíneos. El análisis de los resultados entre las 100 células foliculares pertenecientes a tiroides normal y estimulada revela que existen marcadas diferencias en todos los componentes analizados los que se podría traducir en distintas funcionalidades en el metabolismo de las respectivas glándulas.

Ten 4-month-old Sprague Dawley rats weighing approximately 250 g were divided into two groups of 5 animals each. Group A was the control and the animals in group B received thyroid stimulation with infrared laser in a dose of 16 J/cm2 for 15 consecutive days. Subsequently, rats were euthanized and thyroids were removed and processed for optical microscopy. From both cell types thyroid histological slides and micrographs were obtained with final increases of 400 and 1000X. Morphometric analysis determined the number, areas and cell perimeter as well as colloidal dispersion and presence of blood vessels in 100 follicular cells. Analysis of the results among the 100 follicular cells belonging to normal and stimulated thyroids revealed marked differences in all the analyzed components, which could translate into different functionalities in the metabolism of the respective glands.

Gametogenesis and nucleotypic effects in the tetraploid red vizcacha rat, Tympanoctomys barrerae (Rodentia, Octodontidae).

Nucleotypic effects link DNA content with nuclear size and cell dimensions of reproductive cells in polyploid organisms. We studied the gametogenesis of the allotetraploid rodent Tympanoctomys barrerae, aiming to determine these effects in reproductive cells. The species' cofamily members, Octodon degus and Spalacopus cyanus were used as control. Spermatogenesis and oogenesis in T. barrerae follows the pattern of differentiation and sequence of events of the control species, but varied nucleotypic effects were observed. Exceedingly large, spatulated spermatozoa with a submedially attached flagellum are characteristic of male T. barrerae. The diameter of the nuclei of primordial and growing follicles as well as those of the Graaff follicles, of the granulose, and of luteal cells are significantly larger and heavily heterochromatic. Moreover, the width of the pellucid zone is 108% thicker in T. barrerae than in S. cyanus. Binucleation was recorded in 26% of luteal bodies examined whereas no binucleated cells are detected in the diploid control. Likewise, large heterochromatic nucleoli were observed in the follicle cells but not in S. cyanus. This finding and the high heterochromatin content of reproductive cells in the red vizcacha rat is probably associated with its genome complexity so that redundant genetic information is silenced through heterochromatinization.

Gametogenesis and nucleotypic effects in the tetraploid red vizcacha rat, Tympanoctomys barrerae (Rodentia, Octodontidae)

Nucleotypic effects link DNA content with nuclear size and cell dimensions of reproductive cells in polyploid organisms. We studied the gametogenesis of the allotetraploid rodent Tympanoctomys barrerae, aiming to determine these effects in reproductive cells. The species' cofamily members, Octodon degus and Spalacopus cyanus were used as control. Spermatogenesis and oogenesis in T. barrerae follows the pattern of differentiation and sequence of events of the control species, but varied nucleotypic effects were observed. Exceedingly large, spatulated spermatozoa with a submedially attached flagellum are characteristic of male T. barrerae. The diameter of the nuclei of primordial and growing follicles as well as those of the Graaff follicles, of the granulose, and of luteal cells are significantly larger and heavily heterochromatic. Moreover, the width of the pellucid zone is 108 percent thicker in T. barrerae than in S. cyanus. Binucleation was recorded in 26 percent of luteal bodies examined whereas no binucleated cells are detected in the diploid control. Likewise, large heterochromatic nucleoli were observed in the follicle cells but not in S. cyanus. This finding and the high heterochromatin content of reproductive cells in the red vizcacha rat is probably associated with its genome complexity so that redundant genetic information is silenced through heterochromatinization.

Primer registro de camallanus corderoi Torres, Teuber y Miranda, 1990 (nematoda: camallanidae) en salmónidos cultivados en el lago Puyehue, Chile / First report of camallanus corderoi Torres, Teuber and Miranda, 1990 (nematoda: camallanidae) in cultured salmonids from Puyehue lake, Chile

First report of camallanus corderoi Torres, Teuber and Miranda, 1990 (Nematoda: Camallanidae) in cultured salmonids from Puyehue lake, Chile

Histopatología de kingiella chilenica (bivalvia, cyamiidae) parasitada por redias y cercarias de la familia gymnophallidae / Histopathology of kingiella chilenica (bivalvia, cyamiidae) parasitized by rediae and cercariae of the gymnophallidae family

Se hace un estudio histológico (gónada y hepatopáncreas) en especímenes del microbivalvo kingiella chilenica infectados con larvas de tremátodo digenea, con el fin de definir los órganos en los cuales se localizan las larvas de este parásito. Redias conteniendo cercarias de este tremátodo digenético, aparecen restringidas al tejido hepatopancreático del molusco (estando ausentes en gónadas u otros órganos contiguos) pudiendo mostrar infección masiva de dicho tejido. Por tanto, las gónadas presentan desarrollo gametogénico normal; esto último, unido al hallazgo de hembras parasitadas que igualmente incuban embriones, nos permiten concluir que la infección señalada no afecta, al menos directamente, la actividad reproductiva de este bivalvo