Evaluation of the Glomerular Filtration Barrier by Electron Microscopy

The plasma filtration and formation of the urine is a very complex process necessary for the elimination of metabolites, toxins, and excessive water and electrolytes from the body. The initial process of urine formations is done by the glomerular filtration barrier inside the glomeruli. This specialized barrier consists of three layers, fenestrated endothelium, basement membrane, and podocytes, which ensure that water and small molecules pass through while cells and large molecules are retained. The glomerular filtration barrier is found with abnormal morphology in several diseases and is associated with renal malfunction; thus, it is interesting to study these structures in different experimental and clinical conditions. The normal glomerular barrier and its alterations in some conditions (hypertension, diabetes, and fetal programming) are discussed in this chapter. Furthermore, some methods for studying the glomerular filtration barrier by electron microscopy, both by qualitative and quantitative methods, are present

Immediate and Late Effects of Stress on Kidneys of Prepubertal and Adult Rats

Background/Aims: Stress had been related to physiological and morphological alterations in different organs, however kidney morphology and function after stress conditions are not well-known. To evaluate the effects of chronic stress on renal morphology in prepubertal and adult rats, and to investigate if the effects of induced stress remain stable or worsen following removal of the stress stimuli. Methods: Forty prepubertal rats (4 weeks old) and thirty six adult rats (10 weeks old) were used. Each age group was divided a into control group and a stressed group. Stressed animals were restrained in rigid tubes 2 hours per day for 6 weeks. Animals were euthanized 24 hours after the last stress stimulus for immediate evaluation (50% of the groups), and 6 weeks after the last stress stimulus for late evaluation (50% of the groups). Kidney weight and volume, cortical-medullar ratio, glomerular volumetric density, volume-weighted glomerular volume, and number of glomeruli per kidney were evaluated using the Image J software. Results: Prepubertal stressed animals showed reduced kidney weight and volume, and increased cortical-medullar ratio in comparison to its control group when evaluated immediately. Furthermore, stressed prepubertal and adult animals evaluated immediately had reduced glomerular volumetric density. Most importantly, all stressed groups exhibited reduced number of glomeruli per kidney. Other analyzed parameters did not difeer significantly the groups. Conclusion: Chronic stress induced before and after puberty led to irreversible glomerular loss, however, renal impairment was interrupted by removal of the stress stimuli

Effects of chronic restraint stress in the prostate of prepubertal and adult rats

ABSTRACT Purpose: To investigate the effects of chronic stress in the prostate of prepubertal and adult rats. Methods: Thirty-two male rats were assigned into four groups depending on the type of treatment (control or stressed) and the age at which stress was initiated (prepubertal or adult). Restraint stress stimuli were applied for six weeks. Stressed prepubertal and adult rats evaluated immediately after the last stress stimuli were named SP and SA groups, respectively. Age-matched rats were used as control groups (CP and CA). At the end of the experiment, the rats were euthanized, and prostate morphological parameters were evaluated and statistically compared. Results: Application of stress stimuli to the SP group resulted in reduced body weight, but no prostate morphological modification was noted. The SA group showed reduced testosterone level and prostatic epithelium surface density, in comparison to CA group. Further, the prostatic lumen surface density was increased in adult stressed animals, in comparison to adult controls. Conclusions: The stress stimuli promoted changes in hormonal and morphological parameters in the prostate of adult stressed rats. Prepubertal stressed animals did not presented modifications of prostate morphology

How to Quantify Penile Corpus Cavernosum Structures with Histomorphometry: Comparison of Two Methods

The use of morphometrical tools in biomedical research permits the accurate comparison of specimens subjected to different conditions, and the surface density of structures is commonly used for this purpose. The traditional point-counting method is reliable but time-consuming, with computer-aided methods being proposed as an alternative. The aim of this study was to compare the surface density data of penile corpus cavernosum trabecular smooth muscle in different groups of rats, measured by two observers using the point-counting or color-based segmentation method. Ten normotensive and 10 hypertensive male rats were used in this study. Rat penises were processed to obtain smooth muscle immunostained histological slices and photomicrographs captured for analysis. The smooth muscle surface density was measured in both groups by two different observers by the point-counting method and by the color-based segmentation method. Hypertensive rats showed an increase in smooth muscle surface density by the two methods, and no difference was found between the results of the two observers. However, surface density values were higher by the point-counting method. The use of either method did not influence the final interpretation of the results, and both proved to have adequate reproducibility. However, as differences were found between the two methods, results obtained by either method should not be compared