Joice Naiara Bertaglia Pereira

Effect of caloric restriction on myenteric neuroplasticity in the rat duodenum during aging.

The objective of this study was to evaluate the effects of caloric restriction (CR) on myenteric neurons in the duodenum of Wistar rats during aging. Thirty rats were divided into three groups: the C group (six-month-old animals that were fed a normal diet from weaning until six months of age), the SR group (18-month-old animals that were fed a normal diet from weaning until 18 months of age) and the CR group (18-month-old animals that were fed a 30% CR diet after six months of age). After 12 months, the animals were euthanized. Whole-mount preparations of the duodenums were either stained with Giemsa or underwent NADPH-diaphorase histochemistry to determine the general myenteric neuron population and the nitrergic neuron subpopulation (NADPH-d+), respectively. The NADPH-d-negative (NADPH-d-) neuron population was estimated based on the difference between the Giemsa-stained and NADPH-d+ neurons. The neurons were counted, and the cell body areas were measured. Aging was associated with neuronal loss in the SR group, which was minimized by caloric restriction in the CR group. The density (mm(2)) of the Giemsa-stained neurons was higher in the SR group (79.09 ± 6.25) than in the CR (92.37 ± 11.6) and C (111.68 ± 15.26) groups. The density of the NADPH-d+ neurons was higher in the SR group (44.90 ± 5.88) than in the C (35.75 ± 1.6) and RC (39.14 ± 7.02) groups. The density of NADPH-d- neurons was higher in the CR (49.73 ± 12.08) and C (75.64 ± 17.05) groups than in the SR group (33.82 ± 4.5). In the C group, 32% and 68% of the Giemsa-stained myenteric neurons were NADPH-d+ or NADPH-d-, respectively. With aging (SR group), the percentage of nitrergic neurons (56.77%) increased, whereas the percentage of NADPH-d- neurons (43.22%) decreased. In the CR group, the change in the percentage of nitrergic (42.37%) and NADPH-d- (57.62%) neurons was lower. As NADPH-d- neurons will be mostly cholinergic neurons, CR appears to reduce the loss of cholinergic neurons during aging. The cell body dimensions (μm(2)) were not altered by aging or CR. Thus, CR had a protective effect on myenteric neurons during aging.

Combination of a high-fat diet with sweetened condensed milk exacerbates inflammation and insulin resistance induced by each separately in mice

Obesogenic diets increase body weight and cause insulin resistance (IR), however, the association of these changes with the main macronutrient in the diet remains to be elucidated. Male C57BL/6 mice were fed with: control (CD), CD and sweetened condensed milk (HS), high-fat (HF), and HF and condensed milk (HSHF). After 2 months, increased body weight, glucose intolerance, adipocyte size and cholesterol levels were observed. As compared with CD, HS ingested the same amount of calories whereas HF and HSHF ingested less. HS had increased plasma AST activity and liver type I collagen. HF caused mild liver steatosis and hepatocellular damage. HF and HSHF increased LDL-cholesterol, hepatocyte and adipocyte hypertrophy, TNF-α by macrophages and decreased lipogenesis and adiponectin in adipose tissue (AT). HSHF exacerbated these effects, increasing IR, lipolysis, mRNA expression of F4/80 and leptin in AT, Tlr-4 in soleus muscle and IL-6, IL-1β, VCAM-1, and ICAM-1 protein in AT. The three obesogenic diets induced obesity and metabolic dysfunction. HS was more proinflammatory than the HF and induced hepatic fibrosis. The HF was more detrimental in terms of insulin sensitivity, and it caused liver steatosis. The combination HSHF exacerbated the effects of each separately on insulin resistance and AT inflammatory state.

Benefits of caloric restriction in the myenteric neuronal plasticity in aging rats

Aging is a biologic process characterized by progressive damage of structures and functions of organic systems. In gastrointestinal tract, it can involve enteric nervous system, which plays an important role in digestion and absorption of nutrients, causing hastening of intestinal transit thus reducing its absorptive function. Caloric restriction has been used in several studies with the intention of delaying deleterious effects of aging. This study aimed to evaluate the effects of caloric restriction on myenteric neurons of ileum by aging in rats. 30 Wistar rats were grouped as follows: GI (animals aged 6 months fed with normal diet), GII (animals aged 18 months fed with normal diet) and GIII (animals aged 18 months subject to 31% of caloric restriction). The rats of the GI group were euthanized at 6 months of age and after experimental period of 12 months animals of the group GII and GIII were euthanized, the ileum of all groups were collected, measured and processed by NADPH-dp and Acetylcholinesterase. Quantitative analysis of neurons revealed that aging promotes the increasing of myenteric neurons NADPH-dp and reduces Acetylcholinesterase neuronal population. However, in the cellular profile area, were not observed significant differences between the groups. The caloric restriction has been efficient and can be used preventively because it minimizes quantitative changes associated with aging on ileum myenteric plexuses.

Comparison of Goto-Kakizaki rats and high fat diet-induced obese rats: Are they reliable models to study Type 2 Diabetes mellitus?

Type 2 Diabetes mellitus (T2DM) is an evident growing disease that affects different cultures throughout the world. T2DM occurs under the influence of three main factors: the genetic background, environmental and behavioral components. Obesity is strongly associated to the development of T2DM in the occident, while in the orient most of the diabetic patients are considered lean. Genetics may be a key factor in the development of T2DM in societies where obesity is not a recurrent public health problem. Herein, two different models of rats were used to understand their differences and reliability as experimental models to study the pathophysiology of T2DM, in two different approaches: the genetic (GK rats) and the environmental (HFD-induced obese rats) influences. GK rats were resistant to weight gain even though food/energy consumption (relative to body weight) was higher in this group. HFD, on the other hand, induced obesity in Wistar rats. White adipose tissue (WAT) expansion in this group was accompanied by immune cells infiltration, inflammation and insulin resistance. GK rats also presented WAT inflammation and insulin resistance; however, no immune cells infiltration was observed in the WAT of this group. Liver of HFD group presented fat accumulation without differences in inflammatory cytokines content, while liver of GK rats didn’t present fat accumulation, but showed an increase of IL-6 and IL-10 content and glycogen. Also, GK rats showed increased plasma GOT and GPT. Soleus muscle of HFD presented normal insulin signaling, contrary to GK rats, which presented higher content of basal phosphorylation of GSK-3β. Our results demonstrated that HFD developed a mild insulin resistance in Wistar rats, but was not sufficient to develop T2DM. In contrast, GK rats presented all the typical hallmarks of T2DM, such as insulin resistance, defective insulin production, fasting hyperglycemia/hyperinsulinemia and lipid plasma alteration. Thus, on the given time point of this study, we may conclude that only GK rats shown to be a reliable model to study T2DM.

Evaluation of protein undernourishment on the condylar process of the Wistar rat mandible correlation with insulin receptor expression.

The mandible condylar process cartilage (CP) of Wistar rats is a secondary cartilage and acts as a mandibular growth site. This phenomenon depends on adequate proteins intake and hormone actions, including insulin. Objectives The present study evaluated the morphological aspects and the expression of the insulin receptor (IR) in the cartilage of the condylar process (CP) of rats subjected to protein undernourishment. Material and Methods The nourished group received a 20% casein diet, while the undernourished group (U) received a 5% casein diet. The re-nourished groups, R and RR, were used to assess the effects of re-nutrition during puberty and adulthood, respectively. CPs were processed and stained with picro-sirius red, safranin-O and azocarmine. Scanning electron microscopy and immunohistochemistry were also performed. Results The area of the CP cartilage and the number of cells in the chondroblastic layer decreased in the U group, as did the thickness of the CP layer in the joint and hypertrophic layer. Renourishment during the pubertal stage, but not during the adult phase, restored these parameters. The cell number was restored when re-nutrition occurred in the pubertal stage, but not in the adult phase. The extracellular matrix also decreased in the U group, but was restored by re-nutrition during the pubertal stage and further increased in the adult phase. IR expression was observed in all CPs, being higher in the chondroblastic and hypertrophic cartilage layers. The lowest expression was found in the U and RR groups. Conclusions Protein malnutrition altered the cellularity, the area, and the fibrous cartilage complex, as well as the expression of the IRs.

Balanced Caloric Restriction Minimizes Changes Caused by Aging on the Colonic Myenteric Plexus

ABSTRACT Aging can promote significant morphofunctional changes in the gastrointestinal tract (GIT). Regulation of GIT motility is mainly controlled by the myenteric neurons of the enteric nervous system. Actions that aim at decreasing the aging effects in the GIT include those related to diet, with caloric restriction (CR). The CR is achieved by controlling the amount of food or by manipulating the components of the diet. Therefore, the objective of this study was to evaluate different levels of CR on the plasticity of nicotinamide adenine dinucleotide phosphate– (NADPH-) reactive myenteric neurons in the colon of Wistar rats during the aging process using ultrastructural (transmission electron microscopy) and morphoquantitative analysis. Wistar male rats (Rattus norvegicus) were distributed into 4 groups (n = 10/group): C, 6-month-old animals; SR, 18-month-old animals fed a normal diet; CRI, 18-month-old animals fed a 12% CR diet; CRII, 18-month-old animals fed a 31% CR diet. At 6 months of age, animals were transferred to the laboratory animal facility, where they remained until 18 months of age. Animals of the CRI and CRII groups were submitted to CR for 6 months. In the ultrastructural analysis, a disorganization of the periganglionar matrix with the aging was observed, and this characteristic was not observed in the animals that received hypocaloric diet. It was observed that the restriction of 12.5% and 31% of calories in the diet minimized the increase in density and cell profile of the reactive NADPH neurons, increased with age. This type of diet may be adapted against gastrointestinal disturbances that commonly affect aging individuals.

Quantification and morphometry of myenteric neurones in the jejunum of Holtzman rats (Rattus norvegicus).

With 2 figures and 3 tables

Role of the IPP complex in myelination and remyelination

Autism is one of the most common neurodevelopmental disorders, yet the etiology of the disease remains unknown. Genetic association studies have begun to provide insight into key molecules involved in both autism and the broader autism spectrum disorder (ASD). However, disruption of these candidate genes can only explain the pathology of a small fraction of the total number of affected individuals. A likely explanation for uncovering few monogenetic causes of autism is that the disease is caused by the combination of multiple genetic insults coupled to environmental modifiers. Therefore, current animal models are not sufficiently complex enough to completely mimic the etiology of ASD. In addition, the specific disruption to higher cognition, such as language and social reciprocity, in patients with autism presents a challenge for interpretation of data from non-human systems. To address these difficulties, we examined whether a human neuronal culture system could be utilized in modeling some of the more complex genetic features of autism. These normal human neuronal progenitors (NHNPs) were differentiated into a post-mitotic neuronal state through addition of specific growth factors. We examined whole genome gene expression throughout a time course of differentiation. After 4 weeks of differentiation, the cells displayed both morphological features and gene expression patterns indicative of a neuronal fate. Strikingly, a significant number of genes associated with ASD are either induced or repressed at this time point compared to undifferentiated cells. Moreover, we find a significant percentage of ASD genes highly connected to one another during the differentiation process using an unbiased assessment of underlying gene expression connectivity. Finally, the NHNP cells are genetically tractable, allowing for the manipulation of multiple candidate genes simultaneously or the administration of numerous environmental hazards. Thus, NHNPs can be used to study both the effects of mutation of several ASD candidate genes on neuronal differentiation gene expression and the effects of extracellular molecules. These data should provide us with a better understanding of the signaling pathways disrupted in ASD.