Análise citogenética clássica e molecular em Chelonoidis carbonaria e Phrynops geoffroanus (Testudines)

Os répteis sofreram redução do número de espécies desde a época em que dominavam a Terra até os dias atuais. Os quelônios são poucos estudados, principalmente quanto à sua caracterização citogenética. O presente trabalho objetivou estudar, por meio das técnicas de citogenética clássica e molecular, os cromossomos de Chelonoidis carbonaria, espécie terrestre e Phrynops geoffroanus, espécie ameaçada de extinção que vive nas margens dos rios no continente Sul Americano. As amostras de sangue foram coletadas de animais do criatório, “Reginaldo Uvo Leone”, em Tabapuã-SP. Todos os procedimentos foram aprovados pelo Comitê de Ética Animal (nº 50/07-CEEA – Botucatu – SP) e pelo IBAMA/RAN (nº 14729-1). Para obter as metáfases, o sangue foi inoculado em meio de cultura de linfócitos que foram estimulados a se dividirem pela adição de fito-hemaglutina. Após esse procedimento, a colchicina foi adicionada para inibir a formação das fibras do fuso, mantendo as células em metáfase. Foram avaliados machos e fêmeas de Chelonoidis carbonaria, que apresentaram número de cromossomos igual a 2n = 52, e de Phrynops geoffroanus, descrito pela primeira vez, cujas fêmeas apresentaram 2n = 58 e os machos com 2n = 57 cromossomos. Pela metodologia de bandamento G, foi possível identificar os pares de cromossomos homólogos, e pela técnica de bandamento C, a presença de regiões de heterocromatina constitutiva em macrocromossomos e em microcromossomos. Pela impregnação com íons prata, foram detectadas Regiões Organizadoras Nucleolares (RONs) em dois microcromossomos nos machos e em apenas um microcromossomo nas fêmeas de Chelonoidis carbonaria. O mesmo padrão foi encontrado em Phrynops geoffroanus. A técnica de FISH revelou a presença dos sítios de RNAr marcados na Região Organizadora Nucleolar. O presente estudo permitiu, pelas técnicas de citogenética...The reptiles were reduced in number of species since the time that they ruled the Earth until the present day. The turtles have been little studied, particularly regarding its cytogenetics. The present study investigated, using the techniques of classical cytogenetics and molecular chromosomes of Chelonoidis carbonaria terrestrial species and Phrynops geoffroanus, endangered species that lives along the rivers in the South American continent. Blood samples were collected from animals in the breeding Reginaldo Uvo Leone in Tabapuã-SP. All procedures were approved by the Animal Ethics Committee (nº 50/07-CEEA - Botucatu – SP) and IBAMA/RAN (nº. 14729-1). For the metaphases, the blood was inoculated into the culture medium of lymphocytes that were stimulated to divide by the addition of phyto-hemagglutinin. After this procedure, colchicine was added to inhibit the formation of spindle fibers, maintaining the cells in metaphase. We assessed male and female Chelonoidis carbonaria, which had the same number of chromosomes 2n = 52 and specimens of Phrynops geoffroanus, first described, whose females had 2n = 58 and males with 2n = 57 chromosomes. By G-banding methodology, it was possible to identify pairs of homologous chromosomes and the C banding technique, the presence of regions of constitutive heterochromatic in macrochromosomes and microchromosomes. By impregnation with silver ions were detected Nucleolar Organizer Regions (NORs) in two microchromosomes in males and females in only one of microchromosomes Chelonoidis carbonaria.The same pattern was found in Phrynops geoffroanus. The FISH technique revealed the presence of sites of rRNA marked Nucleolar Organizing Region. This study allowed for the techniques of classical and molecular cytogenetics to identify the chromosomes of males and females of Chelonoidis carbonaria and Phrynops geoffroanus, terrestrial and aquatic... (Complete abstract click electronic access below

Anatomy of the male reproductive system of Phrynops geoffroanus (Testudines: Chelidae) = Anatomia do aparelho reprodutor masculino de Phrynops geoffroanus (Testudines: Chelidae)

The reproductive system of male Phrynops geoffroanus adults ismacroscopically described and the variation in testicular biometry is evaluated. A pair of oval testes is connected by the efferent ductules to the epididymis, which continue as deferent ducts, which emerge in the penis. The volume of the gonads showed the highestaverages during spring and summer months. GSI varied significantly throughout the year, with the highest averages observed in the months that correspond to the end of spring and the beginning of summer, when the reproduction of the species takes place, and the lowest averages were seen in winter, suggesting a cyclical testicular activity.O sistema reprodutor de machos adultos de Phrynops geoffroanus édescrito macroscopicamente e a variação da biometria testicular é avaliada. Um par de testículos ovais está conectado pelos dúctulos eferentes aos epidídimos, que se continuam como ductos deferentes, e que por sua vez desembocam no pênis. O volume das gônadas apresentou as maiores médias durante os meses de primavera e verão. O IGS variousignificativamente ao longo do ano, com maiores médias observadas nos meses que correspondem ao final da primavera e início do verão, quando ocorre a reprodução da espécie, e as menores médias durante o inverno, sugerindo uma atividade testicular cíclica

<b>Anatomy of the male reproductive system of <em>Phrynops geoffroanus</em> (Testudines: Chelidae)</b> - doi: 10.4025/actascibiolsci.v33i4.8091 <b>Anatomy of the male reproductive system of <em>Phrynops geoffroanus</em> (Testudines: Chelidae)</b> - doi: 10.4025/actascibiolsci.v33i4.8091

The reproductive system of male <em>Phrynops geoffroanus</em> adults is macroscopically described and the variation in testicular biometry is evaluated. A pair of oval testes is connected by the efferent ductules to the epididymis, which continue as deferent ducts, which emerge in the penis. The volume of the gonads showed the highest averages during spring and summer months. GSI varied significantly throughout the year, with the highest averages observed in the months that correspond to the end of spring and the beginning of summer, when the reproduction of the species takes place, and the lowest averages were seen in winter, suggesting a cyclical testicular activity.<br>The reproductive system of male <em>Phrynops geoffroanus</em> adults is macroscopically described and the variation in testicular biometry is evaluated. A pair of oval testes is connected by the efferent ductules to the epididymis, which continue as deferent ducts, which emerge in the penis. The volume of the gonads showed the highest averages during spring and summer months. GSI varied significantly throughout the year, with the highest averages observed in the months that correspond to the end of spring and the beginning of summer, when the reproduction of the species takes place, and the lowest averages were seen in winter, suggesting a cyclical testicular activity

Morphological Changes of the Epididymis and Description of the Excurrent Ducts of Phrynops geoffroanus (Testudines: Chelidae) During the Reproductive Cycle

The seminal ducts (efferent ductule, epididymis, and deferent duct) in adults of Phrynops geoffroanus were examined using light microscopy. A series of tubules (efferent ductules) connect the testes to the epididymides. The efferent ductules are formed by a rete of small tubules of varying diameters, with simple columnar epithelium formed by the ciliated cells, nonciliated cells, and few basal cells. The epididymis is a simple, long and highly convoluted tubule that receives the efferent ductules throughout its extension. It is covered by a pseudostratified columnar epithelium with three cellular types: the principal cells, which are the most abundant, basal cells, and a small narrow cell. The histological differences in the epididymis region (cranial, medial, and caudal), as well as the differences in the epithelium throughout the reproductive cycle, are discussed. The deferent ducts consist of a low pseudostratified epithelium with two cellular types: the principal and basal cells. During the months analyzed, spermatozoa were stored in the epididymis, and deferent ducts were found. Anat Rec, 294:145-155, 2011. (C) 2010 Wiley-Liss, Inc.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Association of endothelial nitric oxide synthase (eNOS) gene polymorphisms and physical fitness levels with plasma nitrite concentrations and arterial blood pressure values in older adults.

Endothelial nitric oxide synthase (eNOS) gene polymorphisms are associated with reduced eNOS activity and nitric oxide (NO) production leading to an increase in blood pressure (BP). Regular exercise is the main strategy to minimize the deleterious effects of polymorphisms. However, due to the differences that physical exercise can be performed, some controversial results are found. Therefore it seems reasonable to evaluate the training status (TS). Thus, this study aimed to investigate the association of eNOS gene haplotypes and different levels of TS on nitrite concentrations (NO2-) and BP values in older adult. 424 elderly performed the following assessments: General Functional Fitness Index (GFFI) to estimate TS, systolic and diastolic blood pressure (SBP and DBP), blood collection for analysis of NO2- and g.-786T>C, intron 4b/a (VNTR) and 894G>T polymorphisms. Multivariate logistic regression showed that NO2- was influenced by GFFI and 4b/4a Intron 4. Regarding BP, GFFI influenced SBP and DBP, and just intron 4 was associated with variations in DBP. It can be observed that GFFI affected the NO2-, SBP and DBP independently of haplotypes. Therefore, maintenance of good level of TS can overcome the negative influence of genetics factors (intron 4) by increasing NO2- concentration and decreasing BP values

Association between endothelial nitric oxide synthase and the renin-angiotensin-aldosterone system polymorphisms, blood pressure and training status in normotensive/pre-hypertension and hypertensive older adults: a pilot study

Introduction:Variations in blood pressure (BP) are, in part, genetically determined and some polymorphisms of renin-angiotensin- aldosterone system (RAAS) and synthase of endothelial nitric oxide (eNOS) have been related to hypertension (HT). Conversely, physical exercise is considered a non-pharmacological tool for HT control, treatment, and prevention. Objective: The purpose of this study is to investigate the relationship between eNOS and RAAS polymorphisms, their epistatic interaction, and the respective humoral factors in the BP control in normotensive/pre-hypertension and hypertensive older adults and how this relationship can be modulated by training status (TS) level. Methods:A total of 155 older adults (66.94 ± 6.83 years old) performed the following evaluations: AAHPERD battery test to determine the general functional fitness index (GFFI), systolic and diastolic blood pressure (SBP and DBP), blood collection for DNA extraction, analysis of eNOS gene polymorphisms rs2070744; rs61722009 and rs1799983 and RAAS polymorphisms rs699; rs1799752 and rs5186, and quantification of ACE activity (Fluorimetric Assay) and nitrite concentration (Chemiluminescence Method). Results and Conclusion:Good TS level appears to exert greater influence on SBP for G2 and G3 (G1: 125.79 ± 14.03/ G2: 119.91 ± 11.72/G3: 119.71 ± 10.85) and on NO2 for G3 (G1: 0.42 ± 0.25/ G2: 0.54 ± 0.45/ G3: 0.71 ± 0.52). No associations were observed between eNOS and RAAS polymorphisms, but the epistasis was identified between eNOS polymorphism, rs2070744, and RAAS polymorphism, rs699, revealing a statistically significant interaction (p = .0235) with training score of 0.63, a training test accuracy of 0.61 and a cross-validation consistency of 10/10. This result suggests an increased risk of hypertension

Morphological, morphometrical and ultrastructural characterization of Phrynops geoffroanus' (Testudines: Chelidae) blood cells, in different environments

The aim of this study was to evaluate the formed elements in the periferical blood of two amostral groups of Phrynops geoffroanus: one from an urban environment under domestic sewage dumping, and another from a non-contaminated environment. Blood samples of 36 animals (females and males) were collected through cardiocentesis. Sixteen specimens were from the urban environment, and 20 were from a control environment. Samples of blood tissue were used for light microscopy analysis, and also for morphometric analysis of red blood cells. For the ultrastructural analysis, blood samples of 2 animals were used. The formed elements found, using morphological and ultrastructural analysis were: nucleated red blood cells; thrombocytes; neutrophils, lymphocytes, monocytes, basophils; eosinophils; heterophils, and azurophils. The morphometric analysis of all red blood cells parameters examinated in females showed a statistically significant difference, but in males just the nuclear area showed difference between the specimens of the two environments. The elements identified by light microscopy were elucidated by electron transmission microscopy. This P. geoffroanus study is the first one that makes a correlation between these environments and the description of turtle's blood cells, thus contributing to the identification of the hematological characteristics of this group. (C) 2010 Elsevier Ltd. All rights reserved.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES