Intertubular morphometric and ultrastructural testes analyses in mdx mice

Duchenne Muscular Dystrophy (DMD) reproductive alterations and the influence of antioxidant treatments may aid in understanding morphometry testicular quantification. In this context, the aim of the present study was to characterize the intertubular compartment (ITC) morphometry of animal testes in mdx mice supplemented with ascorbic acid (AA). Sixteen mice were used, namely the C57BL/10 (non-dystrophic) and C57BL/10Mdx (dystrophic) lineages, distributed into the following groups: Control (C60), Dystrophic (D60), Control supplemented with AA (CS60), Dystrophic supplemented with AA (DS60). A total of 200 mg/kg of AA were administered to mice for 30 days. Subsequently, the testicles were collected, weighed, and fragmented. The obtained fragments were fixed in Karnovsky's solution (pH 7.2) and embedded in historesin for morphometric and transmission electron microscopy assessments. Leydig cells were hypertrophic in the D60 group, but was reverted by AA supplementation in the DS60 group. The DS60 group also exhibited increased intertubular volume compared to the CS60 group. The ultrastructural images identified multilamellar bodies in dystrophic animals (lipid storage) and telocyte cells (transport substances) in both control and dystrophic animals. Morphometric alterations were, therefore, noted in the intertubular compartment due to Duchenne muscular dystrophy (DMD), with AA administration capable of altering Leydig cells in this condition.(AU)

Testicular morphometry of rats with Walker 256 tumor supplemented with L-glutamine

Abstract Glutamine is often used to treat metabolic changes associated with anorexia-cachexia syndrome in patients with malignant neoplasms. Walker 256 tumor is an excellent model for studying these changes associated with cancer in different organs, including injuries in testicular functions. However, the effects of supplementing glutamine on testicular morphometry in this model have not yet been investigated. Thus, the objective of this study was to evaluate the effect of L-glutamine supplementation on testicular morphometry in rats transplanted with Walker 256 tumor cells. Forty puberty Wistar rats were divided into four groups: control without L-glutamine (C); control supplemented with L-glutamine (CG); inoculated with Walker 256 tumor cells (WT) and inoculated with Walker 256 tumor cells and supplemented with L-glutamine (WTG). The testicles were removed, weighed, fixed in Bouin, and included in paraffin for histomorphometric analysis. Walker 256 tumor caused quantitative changes in the tubular and intertubular compartments and tunica albuginea, with reductions in the percentages of lumen and tunica albuginea, number of Sertoli cells per gram of testis; number of Leydig cells; percentage of blood vessels and connective tissue in intertubule. However, glutamine supplementation prevented part of these changes caused by the tumor, presenting mainly a protective effect on the tunica albuginea and percentage of blood and lymph vessels in the intertubule. These results indicate the potential of L-glutamine was able to recover for testicular dysfunction associated with cancer.

Testicular morphometry of rats with Walker 256 tumor supplemented with L-glutamine

Glutamine is often used to treat metabolic changes associated with anorexia-cachexia syndrome in patients with malignant neoplasms. Walker 256 tumor is an excellent model for studying these changes associated with cancer in different organs, including injuries in testicular functions. However, the effects of supplementing glutamine on testicular morphometry in this model have not yet been investigated. Thus, the objective of this study was to evaluate the effect of L-glutamine supplementation on testicular morphometry in rats transplanted with Walker 256 tumor cells. Forty puberty Wistar rats were divided into four groups: control without L-glutamine (C); control supplemented with L-glutamine (CG); inoculated with Walker 256 tumor cells (WT) and inoculated with Walker 256 tumor cells and supplemented with L-glutamine (WTG). The testicles were removed, weighed, fixed in Bouin, and included in paraffin for histomorphometric analysis. Walker 256 tumor caused quantitative changes in the tubular and intertubular compartments and tunica albuginea, with reductions in the percentages of lumen and tunica albuginea, number of Sertoli cells per gram of testis; number of Leydig cells; percentage of blood vessels and connective tissue in intertubule. However, glutamine supplementation prevented part of these changes caused by the tumor, presenting mainly a protective effect on the tunica albuginea and percentage of blood and lymph vessels in the intertubule. These results indicate the potential of L-glutamine was able to recover for testicular dysfunction associated with cancer.(AU)

Comparative anatomical description of brain hemisphere surfaces in toninha dolphin and humans

Pontoporia blainvillei (cetacean order and odontoceti suborder), popularly known in Brazil as Toninha, is a small dolphin with a long, thin beak found on the southern shore waters of the western Atlantic. The brain development of dolphins (expressed by the encephalization coefficient) in relation to body size is quite high, and is second only to humans. This high correlation in brain development between dolphins and humans was targeted in this research to describe and compare the morphology of cerebral hemisphere surfaces of these two species. Five brains of Pontoporia blainvillei donated by the BioPesca Project (a non-governmental organization) were analyzed. The brains were fixed in 20% formaldehyde solution for further description and comparison to human brains described in books of neuroanatomy. The telencephalon of Pontoporia blainvillei is rostrally tapered and expanded at the caudal end and it has a greater number of folds/gyrus when compared to human species. Some encephalic grooves/sulcus are similar to grooves found in the human species, such as the cruciate sulcus (central sulcus), lateral fissure (Sylvian cleft), cingulate sulcus and calcarine sulcus. The same encephalic lobes are seen in both species; however the parietal and temporal lobes are highly developed in the Pontoporia blainvillei. These differences are associated with specific function of the species, as the presence of the an efficient auditive system in dolphins.

Comparative anatomical description of brain hemisphere surfaces in toninha dolphin and humans

Pontoporia blainvillei (cetacean order and odontoceti suborder), popularly known in Brazil as Toninha, is a small dolphin with a long, thin beak found on the southern shore waters of the western Atlantic. The brain development of dolphins (expressed by the encephalization coefficient) in relation to body size is quite high, and is second only to humans. This high correlation in brain development between dolphins and humans was targeted in this research to describe and compare the morphology of cerebral hemisphere surfaces of these two species. Five brains of Pontoporia blainvillei donated by the BioPesca Project (a non-governmental organization) were analyzed. The brains were fixed in 20% formaldehyde solution for further description and comparison to human brains described in books of neuroanatomy. The telencephalon of Pontoporia blainvillei is rostrally tapered and expanded at the caudal end and it has a greater number of folds/gyrus when compared to human species. Some encephalic grooves/sulcus are similar to grooves found in the human species, such as the cruciate sulcus (central sulcus), lateral fissure (Sylvian cleft), cingulate sulcus and calcarine sulcus. The same encephalic lobes are seen in both species; however the parietal and temporal lobes are highly developed in the Pontoporia blainvillei. These differences are associated with specific function of the species, as the presence of the an efficient auditive system in dolphins.(AU)

Efeito da atividade física moderada sobre neurônios mioentéricos nitrérgicos do íleo de ratos de meia idade / The effect of moderate physical activity on myoenteric nitrergic neurons in the ileum of middle-aged rats / Efecto de la actividad física moderada en neuronas mientéricas nitrérgicas del íleon de ratas de mediana edad

Poucos são os relatos a respeito do efeito da atividade física sobre o plexo mioentérico ao longo do envelhecimento. Portanto, analisaram-se os neurônios mioentéricos NADPH-diaforase positivos do íleo em ratos que envelheceram praticando atividade física a partir dos seis meses de vida. Dez ratos Wistar foram distribuídos em dois grupos (n = 5/grupo): S (ratos sedentários controles com 12 meses de idade) e T (ratos treinados com 12 meses de idade). Os animais do grupo T realizaram, dos seis aos 12 meses de idade, atividade física regulada conforme resultados obtidos em teste de esforço máximo. Ao final do período experimental, o íleo obtido de cada animal foi processado para a técnica de NADPH-diaforase para evidenciar neurônios mioentéricos em preparados de membrana e quantificar e mensurar a área do pericário desses neurônios. A área do pericário não diferiu (P> 0,05) entre o grupo S (218,49 µm2) e o grupo T (210,59 µm2). A média de neurônios presente em 60 campos microscópicos de preparados de membranas no grupo S (126 neurônios) foi superior (P<0,05) a média encontrada no grupo T (93 neurônios), sugerindo que a atividade física pode inibir ou diminuir a expressão dos neurônios nitrérgicos em ratos de 12 meses de idade que praticaram atividade física desde os seis meses de vida. Os resultados quantitativos sugerem que a falta de atividade física regular propicia aumento na atividade dos neurônios nitrérgicos, contribuindo para o surgimento dos distúrbios da motricidade gastrointestinal que podem ocorrer ao longo do envelhecimento.(AU)

There are only a few reports on the effect of physical activity in the myoenteric plexus throughout the aging process. For such, the positive myoenteric NADPH-diaphoresis neurons in the ileum of those rats that aged practicing physical activity from the sixth month were analyzed. Ten Wistar rats were distributed into two groups (n= 5/group): S (12 months-old control sedentary rats) and T (12 months-old trained rats). The animals from the T group practiced controlled physical activity from the age of 6 months to 12 months, according to the maximum effort test results. At the end of the experimental period, the ileum obtained from each animal was processed in the NADPH-diaphoresis technique to highlight myoenteric neurons in membrane preparations, and quantify and measure the perikaryon area from those neurons. The Perikaryon area did not differ (P>0.05) among the S group (218.49 µm2) and the T group (210.59 µm2). The average of neurons in 60 membrane preparations in the S group (126 neurons) was higher (P<0.05) than the average finding in the T group (93 neurons), indicating that the physical activity can inhibit or decrease the nitrergic expression in 12 month-old rats that have practiced physical activity from the age of 6 months. The quantitative results suggest that the lack of regular physical activity provides the elevation of nitrergic neurons, concurring to the emergence of gastrointestinal motility disorders that can occur during the aging process.(AU)

Hay pocos informes sobre el efecto de la actividad física sobre el plexo mientérico a lo largo del envejecimiento. Por lo tanto, se analizaron las neuronas mientéricas NADPH-diaforasa positivas del íleon en ratas que envejecieron practicando actividad física a partir de seis meses de vida. Diez ratas Wistar se dividieron en dos grupos (n = 5 / grupo): S (ratas sedentarias de control con 12 meses de edad) y T (ratas entrenadas con 12 meses de edad). Los animales del grupo T realizaron, de los seis a los doce meses de edad, actividad física regulada conforme resultados obtenidos en prueba de esfuerzo máximo. Al final del período experimental, el íleon obtenido de cada animal se procesó por la técnica de NADPH-diaforasa para mostrar las neuronas mientéricas en preparados de membrana, cuantificar y medir el área del pericario de esas neuronas. El área del pericario no difirió (P> 0.05) entre el grupo S (218.49 µm2) y el grupo T (210.59 µm2). El número medio de neuronas presentes en 60 campos microscópicos de preparados de membranas en el grupo S (126 neuronas) fue mayor (P <0,05), promedio encontrado en el grupo T (93 neuronas), lo que sugiere que la actividad física puede inhibir o disminuir la expresión de las neuronas nitrérgicas en ratas de 12 meses de edad que practicaron actividad física desde los seis meses de edad. Los resultados cuantitativos sugieren que la falta de actividad física regular promueve aumento en la actividad de las neuronas nitrérgicas, contribuyendo a la aparición de disturbios de la motricidad gastrointestinal que pueden ocurrir a lo largo del envejecimiento.(AU)

Morfometria das fibras colágeno cardíaco em ratos sadios e diabéticos suplementados com vitamina C / Morphometry of the collagen fibers in healthy and diabetic rats treated with vitamin C

The increase of heart collagen fibers in diabetics is a well known fact, but the consequences are not defined. The aim was to quantify the cardiac collagen fibers in normal and diabetic rats treated with vitamin C. We selected 32 Wistar rats, 16 diabetic animals induced endovenously with streptozootocin, and 16 healthy animals, half of them, diabetics and normals, were treated with vitamin C for 90 days. After the experimental proceeding, the hearts were removed and processed accordingly to conventional protocol for optical microscopy and specific staining for collagen. The results showed that the diabetic rats presented increase in the number of cardiac collagen fibers, but the ones treated with vitamin C showed little accumulation of fibers. It could be concluded that treatment with vitamin C is important for the prevention of heart failure in diabetic animals.(AU)

O aumento do conteúdo de fibras colágenas no coração de diabéticos é um fato bastante conhecido, suas conseqüências ainda são objeto de estudo e causam certa controvérsia, portanto este trabalho objetivou estudar a variação na quantidade das fibras de colágeno cardíacas em animais normais e diabéticos tratados pela vitamina C. Para isso foram selecionados 32 ratos Wistar, 16 diabéticos induzidos pela injeção endovenosa de estreptozootocina e 16 normais, sendo metade deles tratados com Vitamina C (diabéticos e normais) por um período de 90 dias. Após período experimental, os corações foram retirados e processados segundo protocolo convencional para microscopia óptica e coloração específica para colágeno. Os resultados mostram que animais diabéticos apresentam maior quantidade de fibras de colágeno cardíacas e que o tratamento com a vitamina C determinou um menor acúmulo na quantidade dessas fibras.(AU)

Morpho-quantitative study of NADH-diaphorase positive myenteric neurons of the ileum of rats of Holtzman lineage (Rattus norvegicus) - DOI: 10.4025/actascibiolsci.v29i3.472 / Estudo morfoquantitativo dos neurônios mioentéricos NADH-diaforase positivos do íleo de ratos (Rattus norvegicus) da linhagem Holtzman - DOI: 10.4025/actascibiolsci.v29i3.472

The myenteric plexus has a regular characteristic morphological pattern for each segment of the digestive tube and for each species of animal. Considering the lack of data pertaining to the mentioned plexus in rats of Holtzman lineage, the objective of this investigation was to carry out a morpho-quantitative study of the myenteric neurons in the ileum, by means of histological sections and whole-mount muscular preparations treated by the NADH-diaphorase method. The profiles of the cell bodies (CB) of the neurons in the mesenteric and antimesenteric regions were counted and measured. The neurons were classified according to the dimensions of the CBs. NADH-dp myenteric neurons were observed grouped together into ganglia in the muscular tunica. The mean neuronal density was 985.8 ± 195.4 neurons/8.96 mm2 in the antimesenteric region and 1267.8 ± 259.92 neurons/8.96 mm2 in the mesenteric region. The incidences of small, medium and large neurons were 14.4, 82 and 3.6% in the antimesenteric region and 14.6, 70.8 and 14.4% in the mesenteric region, respectively. It was concluded that ganglionated arrangements and medium-sized NADH-dp neurons predominated in the myenteric plexus of adult Holtzman rats. The results observed indicated that the NADH-dp myenteric neurons of the ileum of Holtzman rats are similar to those of rats of Wistar lineage with respect to their localization, gangli

O plexo mioentérico possui um padrão morfológico regular característico para cada segmento do tubo digestório e para cada espécie animal. Considerando a escassez de dados pertinentes ao referido plexo em ratos da linhagem Holtzman, o presente estudo como objetivo o estudo morfoquantitativo dos neurônios mioentéricos do íleo, por meio de cortes histológicos e preparados de membrana, tratados pelo método da NADH-diaforase. Foram contados e mensurados os perfis do corpo celular (PC) de neurônios nas regiões mesentérica e antimesentérica. Os neurônios foram classificados segundo as dimensões do PC. Foram observados os neurônios mioentéricos NADH-dp, reunidos em gânglios na túnica muscular. A densidade neuronal média foi de 985,8 ± 195,4 neurônios/8,96 mm2 na região antimesentérica e 1267,8 ± 258,92 neurônios/8,96 mm2 na região mesentérica. As incidências de neurônios pequenos, médios e grandes foram 14,4, 82 e 3,6% na região antimesentérica, e 14,6, 70,8 e 14,4% na região mesentérica, respectivamente. Concluiu-se que, em ratos Holtzman adultos, predominam o arranjo ganglionado no plexo mioentérico e neurônios NADH-dp de tamanho médio. Os resultados encontrados indicam que os neurônios mioentéricos NADH-dp do íleo de ratos Holtzman são similares aos dos ratos da linhagem Wistar no que diz respeito à localização, arranjo ganglionado e predominância de neurônios com PC de tamanho médi

Influência da atividade física sobre o envelhecimento inicial do plexo mioentérico do jejuno em rato Wistar / Influence of the physical activity on the initial aging of myenteric plexus of jejunum in Wistar rat

Embora diversos trabalhos relatem as alterações fisiológicas que ocorrem no trato gastrointestinal com a senescência e com o exercício físico, poucos são os relatos morfológicos a respeito destas duas variantes. Portanto, o objetivo deste trabalho foi verificar qual a influência da atividade física (corrida em esteira) na ultraestrutura, na densidade neuronal (neurônios/mm2) e na área do perfil do corpo celular (µm2) dos neurônios mioentéricos do jejuno de ratos com a idade. Para tanto, foram utilizados 45 ratos Wistar divididos em três grupos: C (controle com seis meses), S (sedentário com 12 meses) e T (treinado com 12 meses) sendo utilizado 5 animais para cada técnica/grupo. Os preparados de membrana do jejuno foram corados com as técnicas histoquímicas de NADH e NADPH para estimar o número de neurônios e a área do perfil neuronal. Os resultados alcançados nos testes de esforço (TEMs) forçam sempre maiores no grupo T do que no grupo S. O peso dos animais do grupo T foi menor que os do grupo S. A área do jejuno-íleo foi mensurada e não presentou diferença significativa (P>0,05) entre os animais dos grupos C, S e T. Com exceção da membrana basal que apresentou-se menos definida e da região periganglionar que encontrava-se mais espassada nos animais do grupo S, não foram identificadas alterações ultraestruturais e no arranjo do plexo mioentéricos entre os grupos. A densidade dos neurônios reativos a NADH-diaforase foi menor (P<0,05) nos animais do grupo S (67,76±3,7) em relação aos animais dos grupos C (104,8±5,86) e T (95,18±7,18). Para os neurônios NADPH-diaforase, observou-se menor densidade neuronal nos grupos S (32,32±1,7) e T (27,39±1,2) comparado com o grupo C (44,53±4,5), mas diferenças significativas não foram evidencias entre os grupos S e T. A área do perfil do corpo celular (µm2) dos neurônios NADH- diaforase positivos diminuiu no grupo S (103,4±8,68) e aumento no grupo T (198,4±8,22) em relação ao grupo C (167±6,93). A área neuronal mensurada nos neurônios NADPH-diaforase positivos foi estatisticamente menor entre os grupos T (129,9±9,55) e C (186,8±9,34), mas não foram observadas diferenças entre estes grupos com o grupo S (157,3±3,64). Contudo, as alterações observadas indicam poucas são as influências do envelhecimento em animais com 12 meses de idade

Although several works tell the physiologic alterations that they happen in the treatment gastrointestinal with the aging and with the physical exercise, few are the morphologic reports regarding these two variants. Therefore, the objective of this work was to verify which the influence of the physical activity (treadmill workout) in the ultrastructural, in the in the neuronal density (neurons/mm2) and in the profile area of the cellular body (µm2) of the myenteric neurons of the jejunum of rats with aging. For so much, 45 Wistar rats were used and they were divided in three groups: C (controls with six months), S (sedentary with 12 months) and T (trained with 12 months) being used 5 animals for each technique/group. The whole mounts of the jejunum were stained with the NADH and NADPH histochemistry techniques to estimate the density and the profile area of the myenteric neurons. The results of the maximal exercise test (METs) they always bigger in the T group than S group. The weight of the animals of the T group was smaller than S group. The jejunum-ileum area was measured and it didn\'t present significant difference (P>0.05) among the animals of the C, s and T groups. Except for the basal membrane that it came less defined and of the periganglionic area that was more scattered in the animals of the S group, they were not identified ultrastructural alterations and in the arrangement of the myenteric plexus among the groups. The density of the NADH-diaphorase neurons was smaller (P<0.05) in the animals of the S group (67.76±3.7) in relation to the animals of the C (104.8±5.86) and T (95.18±7.18) groups. For the NADPH- diaphorase neurons, smaller neuronal density was observed in the S (32.32±1.7) and T (27.39±1.2) groups compared with the C group (44.53±4.5), but the significant differentiate didn\'t evidence among the S and T groups. The profile area (µm2) of the NADH-diaphorase neurons decreased in the S group (103.4±8.68) and increase in the T group (198.4±8.22) in relation to the C group (167±6.93). The profile area of the NADPH-diaphorase neurons was smaller statically among the T (129.9±9.55) and C (186.8±9.34) groups, but differences were not observed among these groups with the S group (157.3±3.64). However, the observed alterations indicate little are the influences of the aging in animals with 12 months age

Avaliação quantitativa e morfométrica dos neurônios mioentéricos nadh-diaforase reativos do estômago de ratos diabéticos induzidos por estreptozootocina e suplementados com ácido ascórbico / Quantitative and morphometric evaluation of nadh-d stained mienteric neurons of the stomach of streptozotocin-induced diabetic rats supplemented with ascorbic acid

A neuropatia autonômica que se manifesta em animais portadores de diabetes mellitus (DM) compromete vários órgãos. A nível gastrointestinal pode acarretar diarréia, constipação, estase e dilatação gástrica, levando a anorexia, perda de peso e vômito. Embora os mecanismos que desencadeiam esses distúrbios não estejam totalmente esclarecidos, parece que alterações do plexo mioentérico estão envolvidas. O estresse oxidativo juntamente com o aumento do sorbitol resultante das alterações metabólicas presentes no DM resultam em edema e lesão neuronal e, entre as conseqüências desta lesão, tem sido descrita alterações nos neurônios mioentéricos. O ácido ascórbico (AA) além da ação antioxidante pode reduzir a concentração de sorbitol atuando, desse modo, como neuroprotetor. O presente trabalho teve como objetivos investigar possíveis alterações no número e no tamanho dos neurônios mioentéricos NADH-diaforase reativos (NADH-dr) do estômago de ratos diabéticos e o efeito da suplementação com AA (1g/L de água) nos referidos parâmetros. Para tanto, 15 ratos (Rattus norvegicus) foram separados em três grupos (n = 5): controle (C); diabético (D); e diabético suplementado com AA (DS). O DM foi induzido por estreptozootocina (35 mg/kg de peso corporal). Após 120 dias de experimento, os animais foram anestesiados, sacrificados e seus estômagos retirados. Os neurônios mioentéricos foram evidenciados pelo método da NADH-diaforase. Por meio de microscópio de luz foram contados os neurônios NADH-dr e, pelo programa computadorizado para análise de imagens, foi mensurado o perfil do corpo celular (PCC) desses neurônios. As regiões aglandular e glandular do estômago foram analisadas separadamente. O número de neurônios NADH-dr foi maior (P<0,05) na região glandular do grupo DS quando comparado com o grupo D. A média dos PCCs foi maior (P<0,05) para os neurônios dos grupos D e DS do que para o grupo C. Ocorreu aumento na incidência de neurônios com PCC superior a 200 µm2 no grupo D quando comparada aos grupos DS e C. Os resultados sugerem que a suplementação com AA teve efeito neuroprotetor sobre os neurônios mioentéricos NADH-dr representado pelo aumento no número de neurônios e diminuição de neurônios grandes na região glandular do grupo DS

The autonomous neuropathy present in animals with diabetes mellitus (DM) compromises several organic systems. On the gastrointestinal tract it may provoke diarrhea, constipation, estase and gastric dilation, leading to anorexia, weight loss and vomit. Although the triggering mechanisms of these disturbances are not completely clear, it seems that alterations of the myenteric plexus may be involved. The oxidizing stress - together with the sorbitol increase resulting from the metabolic alterations due to dm - results in an edema and in a neuronal lesion. Among the consequences of this lesion, alterations in the myenteric neurons have been described. The ascorbic acid (AA), besides its antioxidant effect, may also reduce the sorbitol concentration working as a neuroprotector. The aims of the present work were to investigate the possible alterations in the number and size of nadh-dr stained myenteric neurons of the stomach of diabetic rats and the effect of the aa-supplementation (1g/l of water). In order to do so, 15 rats (rattus norvegicus) were divided into three groups (n = 5): controls (C); diabetic (D); and diabetic supplemented with AA (DS). DM was induced by streptozotocin (35 mg/kg of body weight). 120 days later, the animals were sacrificed for obtaining their stomachs. The myentric neurons were stained employing the nadh-diaforase method. We counted the nadh-dr neurons employing the light microscope and we measured the cellular body profile (CBP) using an image analysis software. The aglandular and glandular areas of the stomach were analyzed separately. The number of neurons nadh-dr was larger (p <0,05) in the glandular area of group ds when compared to group d. The average of cbps was larger (p <0,05) for neurons of groups d and ds than for group c. There was an increase in the incidence of neurons with cbps over 200 mm2 in group D when compared to groups DS and C. The results suggest that the AA supplementation had a neuroprotetor effect on the nadh-dr myenteric neurons represented by the increase in the number of neurons and the decrease of big neurons in the glandular area of group DS