Guía de las actividades de los contenidos del segundo cuatrimestre de la asignatura de Citología e Histología Vegetal y Animal

En este trabajo recogemos las principales pautas y metodología seguida para la elaboración de la Guía de Estudio de parte de los contenidos del segundo cuatrimestre de la asignatura de Citología e Histología Animal y Vegetal de los estudios de Biología. En dicha guía se recoge el desarrollo de las actividades docentes que han sido llevadas a cabo utilizando la plataforma informática Campos Virtual de la Universidad de Alicante. Se trata del ensayo de una nueva manera de abordar el proceso de enseñanza/aprendizaje de nuestros alumnos, tendente a la adaptación de los contenidos de Histología al Espacio Europeo de Educación Superior (EEES). El método empleado ha sido introducir a nuestros alumnos a ser los artífices de su propia formación, mediante la realización tanto de tareas de autoaprendizaje como la exposición de sus resultados al resto de sus compañeros en una serie de seminarios. La metodología empleada se expone en cada uno de los apartados de esta guía.

Metodología y evaluación innovadora en el aprendizaje de dos asignaturas de biología

Introducción de una metodología didáctica diferente a la tradicional, en dos asignaturas troncales de los estudios de Biología: Citología e Histología Vegetal y Animal (primer curso) y Bases Celulares de la Conducta (quinto cuso). Básicamente consiste en proporcionar a los alumnos, previamente y a través de Campus Virtual, los materiales necesarios para su aprendizaje (especialmente tutoriales y ejercicios). La estrategia fundamental es convertir el aula tradicional en un “taller” donde los alumnos, distribuidos en subgrupos de cinco, trabajan con los materiales aportados. Igualmente se les proporciona material de evaluación para que trabajando sobre preguntas, vayan realizando progresivamente una evaluación formativa. Los logros más relevantes fueron: 1) mejores resultados académicos al final del curso; 2) más participación en el aula; 3) mayor aprendizaje autónomo; 4) introducción del alumno en el manejo de enlaces específicos de Internet y en el uso del inglés como lengua científica fundamental, 5) mayor interacción con el profesor a través de las tutorías, tanto virtuales como personales y 6) suscitar una profunda reflexión sobre la excesiva burocratización de la Universidad que interfiere con una gestión ágil e innovadora del conocimiento.

Conocimientos científicos importantes para la conservación y gestión de las tres especies de liebre de la Península Ibérica: deficiencias y retos para el futuro

Los principales avances en el conocimiento de las tres especies de liebre presentes en la Península Ibérica (dos de ellas endémicas) tuvieron lugar a finales del siglo XX con la realización de estudios sobre genética, morfología y ecología descriptiva. Durante los años 90 y 2000, se ha profundizado fundamentalmente en aspectos genéticos y evolutivos, en la definición de factores ambientales determinantes de su distribución, en el estado sanitario y en el comportamiento espacial. En lo que respecta a los estudios sobre dinámica poblacional, se dispone únicamente de trabajos locales con metodologías diversas y series de datos generalmente cortas, lo que impide realizar una interpretación conjunta y sólida de los datos. Esta falta de información conlleva que las bases técnicas para determinar un adecuado aprovechamiento cinegético sean en la mayoría de los casos insuficientes. Además, las tres especies se encuentran fuertemente ligadas al medio agrícola y ganadero, por lo que determinar los factores relativos al manejo de estos medios que afectan a las liebres resulta esencial, especialmente debido al declive general de la mayor parte de las especies silvestres ligadas a estos espacios. ¿Será el sector científico-técnico capaz de avanzar en el conocimiento de la dinámica poblacional y el efecto del manejo agroganadero para una mejor gestión de las liebres ibéricas?

La complejidad de las distrofias hereditarias de la retina: un obstáculo y un reto

ACTO conjunto de FARPE-FUNDALUCE con la SEBBM en el marco de su XXXVII Congreso. Conferencia: La complejidad de las distrofias hereditarias de la retina: Un obstáculo y un reto (José Martín Nieto).

Endophytic colonization of barley (Hordeum vulgare) roots by the nematophagous fungus Pochonia chlamydosporia reveals plant growth promotion and a general defense and stress transcriptomic response

Plant crop yields are negatively conditioned by a large set of biotic and abiotic factors. An alternative to mitigate these adverse effects is the use of fungal biological control agents and endophytes. The egg-parasitic fungus Pochonia chlamydosporia has been traditionally studied because of its potential as a biological control agent of plant-parasitic nematodes. This fungus can also act as an endophyte in monocot and dicot plants, and has been shown to promote plant growth in different agronomic crops. An Affymetrix 22K Barley GeneChip was used in this work to analyze the barley root transcriptomic response to P. chlamydosporia root colonization. Functional gene ontology (GO) and gene set enrichment analyses showed that genes involved in stress response were enriched in the barley transcriptome under endophytism. An 87.5 % of the probesets identified within the abiotic stress response group encoded heat shock proteins. Additionally, we found in our transcriptomic analysis an up-regulation of genes implicated in the biosynthesis of plant hormones, such as auxin, ethylene and jasmonic acid. Along with these, we detected induction of brassinosteroid insensitive 1-associated receptor kinase 1 (BR1) and other genes related to effector-triggered immunity (ETI) and pattern-triggered immunity (PTI). Our study supports at the molecular level the growth-promoting effect observed in plants endophytically colonized by P. chlamydosporia, which opens the door to further studies addressing the capacity of this fungus to mitigate the negative effects of biotic and abiotic factors on plant crops.

Efectos de la erosión del suelo sobre los patrones de la vegetación y su composición florística. Una revisión bibliográfica

En este artículo se revisan los estudios realizados sobre la influencia que tienen los procesos erosivos sobre la vegetación. Fundamentalmente se revisan las tendencias generales en la composición florística de las comunidades vegetales sometidas a fuertes procesos erosivos, así como las tendencias de los patrones de la vegetación y, en menor medida, de los atributos y tipos de plantas. Este campo de trabajo presenta pocos precedentes, siendo escasos los estudios de la influencia de la erosión sobre la vegetación desde un punto de vista ecológico-botánico. Por otro lado, algunos de los resultados parecen a primera vista contradictorios, por lo que es difícil extraer tendencias generales y más o menos universales. Algunas de las generalidades observadas son que el incremento de la erosión del suelo produce un descenso muy claro y mantenido en la cobertura vegetal y en el número de especies. El proceso erosivo no suele acarrear una sustitución de especies vegetales y comunidades, sino solamente la pérdida paulatina de especies, al menos en los estadios más degradados. Por otro lado, se ha observado en ocasiones que la flora de los terrenos más erosionados depende muy fuertemente de las características de la roca madre, variando más entre litologías que la flora de terrenos menos erosionados. Los hemicriptófitos y los caméfitos son las formas vitales de Raunkiaer más frecuentes en estos ambientes. Se discute el papel que pueden tener las diferencias de clima, procesos y tasas erosivas para explicar la gran diversidad de tendencias observadas.

INVES: Docencia en Iniciación a la Investigación en Biología

Un equipo multidisciplinar de profesores y profesoras que imparten docencia en la asignatura Iniciación a la Investigación en Biología, constituyen la Red Docente INVES con el fin de desarrollar una metodología propia de trabajo en equipo, en coordinación con el profesorado de la asignatura Estadística, con la que se comparten objetivos de aprendizaje comunes. Durante el desarrollo de la asignatura, el alumnado diseña y ejecuta un proyecto de investigación bibliométrico de temática biológica sobre un tema actual y de interés. Con ello se favorece la adquisición de competencias transversales del módulo básico del título de grado. La dinámica de trabajo en grupo culmina en la edición de unas Jornadas Científicas, donde los estudiantes exponen los trabajos realizados. Se han consensuado modificaciones en las metodologías y actividades de aprendizaje, mejorando en la eficiencia de la experiencia de enseñanza-aprendizaje.

Tauroursodeoxycholic acid prevents retinal degeneration in transgenic P23H rats

Purpose. To evaluate the preventive effect of tauroursodeoxycholic acid (TUDCA) on photoreceptor degeneration, synaptic connectivity and functional activity of the retina in the transgenic P23H rat, an animal model of autosomal dominant retinitis pigmentosa (RP). Methods. P23H line-3 rats were injected with TUDCA once a week from postnatal day (P)21 to P120, in parallel with vehicle-administered controls. At P120, functional activity of the retina was evaluated by electroretinographic (ERG) recording. The effects of TUDCA on the number, morphology, integrity, and synaptic connectivity of retinal cells were characterized by immunofluorescence confocal microscopy. Results. The amplitude of ERG a- and b-waves was significantly higher in TUDCA-treated animals under both scotopic and photopic conditions than in control animals. In the central area of the retina, TUDCA-treated P23H rats showed threefold more photoreceptors than control animals. The number of TUNEL-positive cells was significantly smaller in TUDCA-treated rats, in which photoreceptor morphology was preserved. Presynaptic and postsynaptic elements, as well as the synaptic contacts between photoreceptors and bipolar or horizontal cells, were preserved in TUDCA-treated P23H rats. Furthermore, in TUDCA-treated rat retinas, the number of both rod bipolar and horizontal cell bodies, as well as the density of their synaptic terminals in the outer plexiform layer, was greater than in control rats. Conclusions. TUDCA treatment was capable of preserving cone and rod structure and function, together with their contacts with their postsynaptic neurons. The neuroprotective effects of TUDCA make this compound potentially useful for delaying retinal degeneration in RP.

Rotenone induces degeneration of photoreceptors and impairs the dopaminergic system in the rat retina

Rotenone is a widely used pesticide and a potent inhibitor of mitochondrial complex I (NADH-quinone reductase) that elicits the degeneration of dopaminergic neurons and thereby the appearance of a parkinsonian syndrome. Here we have addressed the alterations induced by rotenone at the functional, morphological and molecular levels in the retina, including those involving both dopaminergic and non-dopaminergic retinal neurons. Rotenone-treated rats showed abnormalities in equilibrium, postural instability and involuntary movements. In their outer retina we observed a loss of photoreceptors, and a reduced synaptic connectivity between those remaining and their postsynaptic neurons. A dramatic loss of mitochondria was observed in the inner segments, as well as in the axon terminals of photoreceptors. In the inner retina we observed a decrease in the expression of dopaminergic cell molecular markers, including loss of tyrosine hydroxylase immunoreactivity, associated with a reduction of the dopaminergic plexus and cell bodies. An increase in immunoreactivity of AII amacrine cells for parvalbumin, a Ca2+-scavenging protein, was also detected. These abnormalities were accompanied by a decrease in the amplitude of scotopic and photopic a- and b-waves and an increase in the b-wave implicit time, as well as by a lower amplitude and greater latency in oscillatory potentials. These results indicate that rotenone induces loss of vision by promoting photoreceptor cell death and impairment of the dopaminergic retinal system.

Safranal, a saffron constituent, attenuates retinal degeneration in P23H rats

Saffron, an extract from Crocus sativus, has been largely used in traditional medicine for its antiapoptotic and anticarcinogenic properties. In this work, we investigate the effects of safranal, a component of saffron stigmas, in attenuating retinal degeneration in the P23H rat model of autosomal dominant retinitis pigmentosa. We demonstrate that administration of safranal to homozygous P23H line-3 rats preserves both photoreceptor morphology and number. Electroretinographic recordings showed higher a- and b-wave amplitudes under both photopic and scotopic conditions in safranal-treated versus non-treated animals. Furthermore, the capillary network in safranal-treated animals was preserved, unlike that found in untreated animals. Our findings indicate that dietary supplementation with safranal slows photoreceptor cell degeneration and ameliorates the loss of retinal function and vascular network disruption in P23H rats. This work also suggests that safranal could be potentially useful to retard retinal degeneration in patients with retinitis pigmentosa.

Gene cloning, molecular modeling, and phylogenetics of serine protease P32 and serine carboxypeptidase SCP1 from nematophagous fungi Pochonia rubescens and Pochonia chlamydosporia

The fungi Pochonia chlamydosporia and Pochonia rubescens are parasites of nematode eggs and thus are biocontrol agents of nematodes. Proteolytic enzymes such as the S8 proteases VCP1 and P32, secreted during the pathogenesis of nematode eggs, are major virulence factors in these fungi. Recently, expression of these enzymes and of SCP1, a new putative S10 carboxypeptidase, was detected during endophytic colonization of barley roots by these fungi. In our study, we cloned the genomic and mRNA sequences encoding P32 from P. rubescens and SCP1 from P. chlamydosporia. P32 showed a high homology with the serine proteases Pr1A from the entomopathogenic fungus Metarhizium anisopliae and VCP1 from P. chlamydosporia (86% and 76% identity, respectively). However, the catalytic pocket of P32 showed differences in the amino acids of the substrate-recognition sites compared with the catalytic pockets of Pr1A and VCP1 proteases. Phylogenetic analysis of P32 suggests a common ancestor with protease Pr1A. SCP1 displays the characteristic features of a member of the S10 family of serine proteases. Phylogenetic comparisons show that SCP1 and other carboxypeptidases from filamentous fungi have an origin different from that of yeast vacuolar serine carboxypeptidases. Understanding protease genes from nematophagous fungi is crucial for enhancing the biocontrol potential of these organisms.

Calcineurin expression and activity is regulated by the intracellular redox status and under hypertension in human neutrophils

Calcineurin (protein phosphatase 2B) (CN) comprises a family of serine/threonine phosphatases that play a pivotal role in signal transduction cascades in a variety of cells, including neutrophils. Angiotensin II (Ang II) increases both activity and de novo synthesis of CN in human neutrophils. This study focuses on the role that intracellular redox status plays in the induction of CN activity by Ang II. Both de novo synthesis of CN and activity increase promoted by Ang II were downregulated when cells were treated with l-buthionine-(S,R)-sulfoximine, an inhibitor of synthesis of the antioxidant glutathione. We have also investigated the effect of pyrrolidine dithiocarbamate and phenazine methosulfate, which are antioxidant and oxidant compounds, respectively, and concluded that the intracellular redox status of neutrophils is highly critical for Ang II-induced increase of CN expression and activity. Results obtained in neutrophils from hypertensive patients were very similar to those obtained in these cells on treatment with Ang II. We have also addressed the possible functional implication of CN activation in the development of hypertension. Present findings indicate that downregulation of hemoxygenase-1 expression in neutrophils from hypertensive subjects is likely mediated by CN, which acts by hindering translocation to the nucleus of the transcription factor NRF2. These data support and extend our previous results and those from other authors on modulation of CN expression and activity levels by the intracellular redox status.

Transcription of liver X receptor is down-regulated by 15-deoxy-Δ12,14-prostaglandin J2 through oxidative stress in human neutrophils

Liver X receptors (LXRs) are ligand-activated transcription factors of the nuclear receptor superfamily. They play important roles in controlling cholesterol homeostasis and as regulators of inflammatory gene expression and innate immunity, by blunting the induction of classical pro-inflammatory genes. However, opposite data have also been reported on the consequences of LXR activation by oxysterols, resulting in the specific production of potent pro-inflammatory cytokines and reactive oxygen species (ROS). The effect of the inflammatory state on the expression of LXRs has not been studied in human cells, and constitutes the main aim of the present work. Our data show that when human neutrophils are triggered with synthetic ligands, the synthesis of LXRα mRNA became activated together with transcription of the LXR target genes ABCA1, ABCG1 and SREBP1c. An inflammatory mediator, 15-deoxy-Δ12,14-prostaglandin J2 (15dPGJ2), hindered T0901317-promoted induction of LXRα mRNA expression together with transcription of its target genes in both neutrophils and human macrophages. This down-regulatory effect was dependent on the release of reactive oxygen species elicited by 15dPGJ2, since it was enhanced by pro-oxidant treatment and reversed by antioxidants, and was also mediated by ERK1/2 activation. Present data also support that the 15dPGJ2-induced serine phosphorylation of the LXRα molecule is mediated by ERK1/2. These results allow to postulate that down-regulation of LXR cellular levels by pro-inflammatory stimuli might be involved in the development of different vascular diseases, such as atherosclerosis.

The ubiquitin-proteasome system in retinal health and disease

The ubiquitin–proteasome system (UPS) is the main intracellular pathway for modulated protein turnover, playing an important role in the maintenance of cellular homeostasis. It also exerts a protein quality control through degradation of oxidized, mutant, denatured, or misfolded proteins and is involved in many biological processes where protein level regulation is necessary. This system allows the cell to modulate its protein expression pattern in response to changing physiological conditions and provides a critical protective role in health and disease. Impairments of UPS function in the central nervous system (CNS) underlie an increasing number of genetic and idiopathic diseases, many of which affect the retina. Current knowledge on the UPS composition and function in this tissue, however, is scarce and dispersed. This review focuses on UPS elements reported in the retina, including ubiquitinating and deubiquitinating enzymes (DUBs), and alternative proteasome assemblies. Known and inferred roles of protein ubiquitination, and of the related, SUMO conjugation (SUMOylation) process, in normal retinal development and adult homeostasis are addressed, including modulation of the visual cycle and response to retinal stress and injury. Additionally, the relationship between UPS dysfunction and human neurodegenerative disorders affecting the retina, including Alzheimer's, Parkinson's, and Huntington's diseases, are dealt with, together with numerous instances of retina-specific illnesses with UPS involvement, such as retinitis pigmentosa, macular degenerations, glaucoma, diabetic retinopathy (DR), and aging-related impairments. This information, though still basic and limited, constitutes a suitable framework to be expanded in incoming years and should prove orientative toward future therapy design targeting sight-affecting diseases with a UPS underlying basis.