LABORATORY GUIDE FOR URINARY SYSTEM


EXTRARENAL COLLECTING SYSTEM

Slide #13 was taken from a human kidney which underwent autolysis and is less than ideal for learning histologic features. It is useful to view this section grossly holding the slide against a white background to distinguish the lobated pattern of the human kidney. Note several papillae and minor calyces.

Slide #107 shows proximal and distal sections of a ureter. Identify the layers, noting especially the transitional epithelium. How can you tell which one was nearest the kidney and which one was nearest the bladder? Also, do you see a serosa?

Slide #14 is a section of bladder with a piece of ureter included in the lumen strangely enough. Review all layers of both structures. The bladder epithelium appears better preserved than that of the ureter. Note the dense irregular connective tissue of the lamina propria, and in the bladder, its vascularity. How many muscle layers are there in the muscularis? Is there a serosa present?

KIDNEY

Zones

This is best performed on the perfused unilobate rat kidney found on Slide 89. (If you want you can do this after you've identified the different portions of the nephron.) First examine the kidney grossly and then with an inverted ocular to locate cortex, outer medulla (outer zone), and inner medulla (inner zone). This will be fun and challenging since the sections distributed through the class have many different orientations and, in some cases, don't have all zones included. If you find you've been deprived of a zone, harass your immediate neighbors for their kidneys. The ideal section is shown below in cut A. Less ideal sections occur with cuts that are superficial or oriented strangely, shown in cut B.

Arcuate veins will appear as large, empty circular profiles and serve to delineate cortex from medulla, which can't be distinguished grossly on the basis of texture or color. The medulla is subdivided into several zones and stripes. The outer zone of the medulla is clearly distinguishable from the inner zone, which is light and has a fine texture. The inner zone is the portion referred to grossly as the papilla. The outer zone is further subdivided into stripes. The outer stripe is very eosinophilic due to the presence of large, eosinophilic proximal tubules. The inner stripe paler due to absence of proximal tubules. Thus, while the outer medulla is two-toned with its two stripes, the inner medulla appears as a more homogeneous area.

Nephron Segments: Slide #89 (rat) and 108 (human)

Most portions of the nephron are more easily identified with the perfused rat kidney than the human kidney. The proximal tubule is especially sensitive to fixation artifact and can only be seen as it would appear in life in the perfused kidney. Thus in this isolated situation you will find a non-human tissue being used in this histology course. However, as you will be seeing human kidneys during the course of your clinical practice, it is imperative to be familiar with the nephron as it appears in the human also. Since rat and human nephron sections are so similar, you should be able to identify either except in a few places noted below where rat will not be used for testing purposes.

Begin your study of the nephron in the inner medulla (papilla), and work your way out to the cortex. The medulla has the simplest structure and fewer nephron types, whereas the cortex is more complex.

Even to the experienced, not all tubule types are identifiable in paraffin sections! If some tubules are indeterminant, as many will be, don't concern yourself, but concentrate on being able to identify good examples of each structure underlined.

Inner Medulla (Inner Zone of the Medulla)
Slide 108 of human kidney has excellent examples of collecting ducts. These are the largest circular profiles with large lumens and a simple tall columnar lining epithelium.. At the tip of the papilla, these ducts can be seen in longitudinal section opening into a minor calyx. Other small circular profiles represent thin limbs of Henle and vasa rectae, which can't be distinguished with certainty unless you see erythrocytes in the lumen of the latter. These are both lined by a simple squamous epithelium. These thin-walled structures may either be descending or ascending, but we can't distinguish either in these paraffin sections. The interstitium fills in between these structures.

In the rat kidney, the collecting ducts are lined by a flatter epithelium, and the best way to identify them is by their wide lumens. You will not be tested on the appearance of rat collecting duct since it is not representative of human histology.

Outer Medulla (Outer Zone of the Medulla)
Inner Stripe
The border between inner and outer medulla is sharply divided by the sudden presence in the inner stripe of the latter of thick ascending limbs. Thus to identify these positively, go to this border and note tubules lined by a cuboidal epithelium which is slightly more eosinophilic than that of the collecting ducts.

Thin limbs and capillaries again are difficult to distinguish from each other, but tend to aggregate together in bundles, separate from the thick ascending limbs. As usual, they are lined by a squamous epithelium.

Look for collecting ducts as they course through this stripe.

Outer Stripe
In the outer stripe of the outer medulla, the straight part of the proximal tubule appears abruptly. These are very eosinophilic and are the tubule type with a brush border. They are cuboidal to low columnar. Look carefully for a profile showing the transition from a proximal tubular epithelium to the squamous epithelium of the thin descending limb. This is the only site you will definitely be able to identify a thin limb, and furthermore, to state whether it is ascending or descending. Thick ascending limbs are still coursing out to the cortex, but are being lined by a progressively lower epithelium than was found in the inner stripe. This will continue as they return to the glomerulus. Collecting ducts continue here, but are lined by a lower epithelium than in the inner medulla.

Cortex
This region of the kidney is defined by the presence of renal corpuscles, and is "separated" from outer medulla by arcuate vessels. Distinguish arcuate arteries from arcuate veins, which are extremely thin-walled for vessels of this size; frequently displaying only a lining epithelium at this level of resolution.

Identify the components of the renal corpuscle: Look for afferent/efferent arterioles - you can't tell them apart, but you can see them entering Bowman's capsule at the vascular pole. Find the glomerulus, parietal and visceral layers of Bowman's capsule separated by Bowman's space. At the urinary pole the parietal epithelium abruptly changes to the tall epithelium of the proximal tubule. Identify glomerular capillaries, and nuclei of podocytes (remember these are the cells of the visceral epithelium) vs. endothelial cells. Identify intraglomerular and extraglomerular mesangial cells. The latter can only be appreciated when both entering and exiting arterioles are in the plane of section at the vascular pole. These extraglomerular mesangial cells will be found between them.

Look for proximal tubules in the rat kidney first, focusing up and down to resolve the brush border. These tubules will appear collapsed and fuzzy in the human kidney, and certainly a brush border is not resolvable. Convoluted and straight segments appear identical, but the straight, or pars recta, sections will be seen coursing in long rays parallel to thick ascending limbs and collecting ducts. Cells of proximal tubules are low columnar to cuboidal; their nuclei are located in a central position in each cell.

Whereas the thick ascending limb displays a tall cuboidal epithelium in the outer medulla, as it approaches the glomerulus it becomes low cuboidal to high squamous. Search the corpuscles for fortuitous sections showing an approaching thick ascending limb. At a variable distance from the glomerulus, the epithelium makes a quick transition to a cuboidal epithelium again. This is the beginning of the distal convoluted tubule. Distal convoluted tubules lack a brush border and have dense, apically-located nuclei.

Be sure to find a macula densa. This will be seen as a cluster of cells in the thick ascending limb where it passes between the afferent and efferent arterioles, forming the juxtaglomerular apparatus. In the human, these cells are cuboidal to columnar and tightly packed. In the rat the cells of the macula densa are flatter. In either case, the close packing of the cells causes their nuclei to form a characteristic dense area, giving rise to the term macula densa, or dense spot.

To find cortical collecting ducts, look for longitudinally oriented tubules running in rays perpendicular to the surface of the kidney. The epithelium of these tubules in the cortex appears fairly flat, with a wide lumen, thus do not expect them to look like they do in the inner medulla. The hallmark of the cortical collecting duct is the presence of two cell types. Most of the lining cells will be flat--these are the principal cells. Interspersed among these are plump intercalated cells.

The space between the tubules is the interstitium. It is quite sparse and occupied primarily by peritubular capillaries.