Anatomy and Cell Biology 3309

ENDOCRINE SYSTEM (I) The endocrine system has the following glands: adrenal, thyroid, parathyroids, pituitary, pineal body, part of testes, part of ovaries, enteroendocrine system, pancreatic islets. Only the first five will be discussed in detail, the others are covered with the study of the reproductive system and the digestive system. The products of the endocrine glands are the hormones which are secreted into a highly vascular connective tissue from which they diffuse into the blood present in the capillaries. There is, therefore, a close association of the capillary system with the endocrine glands. The endocrine glands may also be called ductless glands as, unlike in exocrine glands, there is no duct to deliver the hormones to the surface. Each type of endocrine gland is quite unique in the method of delivery of the hormones into the blood. Therefore, the structural aspects of secretion, and the capillary-gland association have to be learned for each gland separately. A common feature of the endocrine glands is that they derive from the embryonic epithelium, originally from an invagination which eventually sinks into the underlying connective tissue, transforms into a secretory gland and looses its former connection with the surface epithelium from which it derives. Adrenal gland First view the adrenal with the low power objective using slide 96. Identify: capsule, cortex, medulla, medullary veins, z. glomerulosa, z. fasciculata, z. reticularis. Now examine your slide with the low power objective. First examine the capsule. Note that it is rather well developed and contains dense connective tissue. In the zona glomerulosa, notice that the cells are arranged in arcades or clumps. Some of the cells are vacuolated indicating former sites of lipid (steroid) droplets. Select a typical part of the zona fasciculata. Identify the cell cords which are more or less straight and the sinusoidal capillaries, with their endothelium, in between the cords. The relative area occupied by the various layers may change from slide to slide and also within a section depending on the plane of sectioning. The next layer to be examined is the zona reticularis. Notice that the cell cords run rather irregularly in comparison to the zona fasciculata. Examine now the details. The thin zona glomerulosa lies beneath the capsule and consists of groups of cells separated by thin connective tissue trabeculae derived from the capsule. The zona fasciculata is thick and there is a tendency for the cells to be arranged in radial cords. The inner zona reticularis is of intermediate thickness and the irregularly disposed cords of cells anastomose with each other. Look at the various zones with the high power objectives, confirming the general arrangement of the cells. Now examine the zona glomerulosa with the high power objective. Each glomerulus is made up of cuboidal or columnar cells. The nuclei are circular or oval in outline and have a moderately large nucleolus. The cytoplasm is pale and vacuolated. Part of the vacuolation is due to lipids which have been removed during preparation of the tissue, although the lipid content is considerably higher in the next zone. There is a fairly abrupt transition between the zona glomerulosa and the zona fasciculata. Still using the high power lens, the cells of the zona fasciculata will be seen to have an irregularly polyhedral shape and approximately spherical nuclei. The vacuolated appearance of the cytoplasm (due to dissolved lipid droplets) is so marked that these cells are sometimes called spongiocytes. In the zona reticularis the cells are also polyhedral and the cytoplasm stains darkly. There are often signs of cell degeneration in this zone, such as shrunken nuclei with abnormal staining properties. There is probably a general tendency toward new formation of cells in the outer part of the cortex, a slow movement of cells centrally, and cell degeneration in the zona reticularis. The adrenal cortex elaborates hormones, belonging chemically to the steroid group of substances, which have a vital effect on various aspects of metabolism. For example, deoxycorticosterone controls salt balance, cortisone influences protein and carbohydrate metabolism and others have an effect similar to that of the sex hormones. The lipid droplets of the cells are an indication of the secretory activity of the adrenal

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Anatomy and Cell Biology 3309

cortex. Using the low power objective, the adrenal medulla should now be examined in slide 96. Using the high power objective, the large number of capillaries in addition to venules will be seen. The cells tend to be arranged in round groups or short cords with cell outlines sometimes seen. The cytoplasm presents no distinguishing features, however if the medulla is treated with chromium salts, fine brown granules (chromaffin granules) may be seen in the cytoplasm. Before closing the study of the adrenal gland, review the main features of the EM structure of the cortical cells and the blood circulation pattern of the gland. Thyroid gland In slides 52 and 57, locates the thyroid gland. The colloid of the follicles is conspicuously red because of the PAS staining. The size of the follicles vary mainly because the plane of sectioning is different for each of these spherical structures. There is also a natural variation in size. Select a follicle which shows well preserved colloid and follicular cells (high power view). Note that the follicles fit together snugly. They are separated only by a little reticular connective tissue which contains a rich capillary network. The boundaries of individual cells are indistinct but the number of cells in the epithelium is indicated clearly by the nuclei. These are the important secretory cells. Look for PAS positive resorption granules in the supranuclear cytoplasm. The follicle contains viscous PAS positive colloid, a product of the follicular epithelium. The colloid fills the follicles in life. Peripheral vacuoles may be seen. This is not entirely artefact, since the peripheral vacuolation indicates a difference in the physical properties of the colloid at the periphery of the follicle. The cells of the follicular epithelium obtain basic products (especially iodine, tyrosine and other amino acids) from the blood capillaries, and synthesize thyroglobulin which passes into the follicle to form the chief component of the colloid. The iodine becomes oxidized into iodide in the cells and this passes into the colloid where the thyroglobulin is converted into iodothyroglobulin. Part of the stored colloid is resorbed by the follicular cell where enzymes reduce the large molecule of iodothyroglobulin to smaller and more diffusible components, such as thyroxine, which enter parafollicular connective tissue and then the blood stream as the physiological need of the body demands. A section through the human thyroid and parathyroid glands is present on slide 55. This slide has been stained with H & E. Note that the size of the thyroid follicles is much greater than in the previous slide. Identify some of the structures between the follicles. The parafollicular cells that are associated with the follicular epithelial cells can only be visualized with special techniques (see demonstration). Parathyroids The parathyroid gland may be studied using slides 55 and 57. First, locate the gland with the low power objective. The gland is associated closely with the thyroid gland in most mammalian species. Identify the capsule of the gland and then notice that the substance of the gland consists of very small cells most of which are the chief cells. Oxyphil cells are fewer in comparison to the chief cells and often are found in clumps, so that the whole gland must be searched to find them if present at all. When a group of oxyphil cells is seen, study them with the oil immersion objective and note the relatively large amount of eosinophilic cytoplasm and the denser nucleus than one sees in chief cells. Oxyphil cells may also be seen in the demonstration material. Some mammalian species do not have oxyphil cells in the parathyroid gland. In the human, the oxyphil cell makes its appearance around puberty. What is the main ultrastructural feature of the oxyphil cell? Review the functional features of the thyroid and parathyroid glands. It is important to know how the various structural elements participate in the elaboration of the hormones.

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ENDOCRINE SYSTEM (II) Pituitary Gland First, study slide 77 under the lower power. This is a pituitary gland from the cat. Knowledge of terminology is important (see textbook and also the demonstration material). When you view the pituitary with the low power objective, essentially 3 major areas may be seen: pars distalis (pars anterior), pars intermedia and pars nervosa. Now look at slide 54, a human pituitary, with the low power objective and note the differences. The pars intermedia is more difficult to identify and is much less developed than in the cat. Using slide 77, note that there is a cleft between the pars distalis and the pars intermedia. The pars intermedia is cellular with most cells basophilic, consequently the overall appearance is different from the pars distalis. Also, the pars intermedia seems to envelop the pars nervosa in the cat. In the human pituitary, (slide 54 and demonstration material) the pars intermedia is much less developed, the cleft is difficult to see and there are large follicle structures containing a colloid that occupy most of the pars intermedia. The pars tuberalis (not well shown in most of the sections) is basically an upward extension of the pars distalis. The overall appearance is similar to that of the pars distalis but the cellular details are different. The infundibulum is basically an upward extension of the pars nervosa. Using also slide 54, examine the pars distalis. Note that the cells are in cords with wide sinusoidal capillaries between the cords. Examine the cellular details of the pars distalis under high power. About 50% of the cells stain poorly. These cells are the chromophobes. Note that in certain areas either acidophils or basophils seem to predominate. The acidophilic cell population contains the somatotrophs and the mammotrophs. These two cell types cannot be distinguished with routine stains. The basophilic cells are the thyrotrophic, gonadotrophic and the corticotrophic cells. The thyrotrophs tend to be larger and angular and the gonadotrophs smaller and round. The ACTH producing cells are probably of similar appearance to the thyrotrophs. The pars nervosa has a very characteristic appearance in the pituitary. Most of the pars nervosa consists of nerve fibers. The nuclei that are seen to be scattered in the tissue belong mainly to pituicytes. With the H & E stain, Herring bodies may be seen, however, when special staining methods are used (demonstration material). Herring bodies are shown to better advantage.

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The details of the pituitary gland should be studied first in the cat (slide 77). The general mammalian plan of the pituitary is well illustrated in this species. The most informative sections are those through the midline of the pituitary, although all will not conform to this optimal condition. First, note the pale portion of the gland which is the part derived embryologically from the brain. The main mass of the neural derivative is the pars nervosa. The pars nervosa extends upwards as the infundibulum or pituitary stalk which joins the floor of the hypothalamic part of the brain. In midline sections, an extension of the third infundibular recess is closed in the mature human pituitary. With this information, the section may be oriented in the proper vertical direction. The large dense mass of cells in front of the pars nervosa is the pars distalis (anterior). It is a development of the anterior wall of Rathke's pouch. The cleft behind the pars anterior is derived from the lumen of Rathke's pouch. It is represented only by small irregular spaces in the mature human pituitary. The layer of cells intervening between the cleft just mentioned and the pars nervosa is the pars intermedia, a derivative of the posterior wall of Rathke's pouch. There is an extension of the pars anterior, known as the pars tuberalis, upwards around the infundibulum and even on the under surface of the hypothalamus. The pars tuberalis is probably not be included in the section now being studied. Now study the cell details of the pituitary in the same section, first in the anterior lobe. Three types of gland cells can be distinguished histologically chromophobes, basophils, acidophils. Only the latter two of these are actively secreting. With the low power objective, the pars distalis has the appearance of an almost solid mass of cells. With the high power objective, it will be noted that the cells are arranged in irregular cords or masses. The intervening sinusoids (the pars distalis is very vascular) are lined in by endothelium. The most conspicuous cells can be distinguished from the remainder by virtue of the densely packed eosinophilic granules in their cytoplasm. These are the acidophiles. For more cytological detail use the high power objective. The three types of gland cells may be recognized as follows. The most numerous (about 50%), but not the most conspicuous cell is the chromophobe cell). This cell is of moderate size, and its round to oval vesicular nucleus contains a small nucleolus. The cytoplasm is pale and contains no granules. It is thought that the chromophobes are resting cells, in so far as hormone secretion goes, and that they can transform into one or other of the remaining cell types. The other half of the gland cell population is made up of chromophils, so called because the cytoplasm has a stronger affinity for dyes. By far the more conspicuous variety is the acidophil, which is distinguished by a cytoplasm containing many minute eosinophilic granules. The acidophil is larger than a chromophobe; its nucleus is spherical or irregular and contains a nucleolus. Acidophil make up nearly 40% of the cell population and are actively secreting cells. The basophil is the other type of chromophil. The basophils are also less frequent than the other cell types, making up only about 10% of the cell population. With careful searching, clumps of basophils may be found and recognized by their usually large size (most a little larger than acidophils and considerably larger than chromophobes) and by their basophilic granules. They, like the acidophils, are actively engaged in the elaboration of hormones. Now examine the pars intermedia under oil immersion. This portion of the pituitary is better developed in the cat than in man. Note the almost solid mass of fairly large cells with spherical to oval vesicular nuclei and lightly basophilic cytoplasm. No function has been discovered for these cells in mammals although, in amphibia, a hormone of the pars intermedia (intermedin) has an effect on pigment-bearing cells (or chromatophores) of the skin. Turn now, with the high power lens, to the pars nervosa and an entirely different histological picture will be seen. Cell outlines are invisible but there are scattered nuclei, varying in size and shape, and faintly fibrillated background material. The cells are the pituicytes, similar to the neuroglial or supporting cells of the brain. Look at the demonstration slide showing neurosecretory substance.

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Pineal Gland Study the pineal gland in slide 53. Like the pars nervosa of the pituitary, the pineal body gives an incomplete picture in sections stained with H & E. With the low power objective, note the arrangement of the cells in whorls or cords separated by thin septa of connective tissue. Search the section for irregular masses stained deeply with haematoxylin. These calcareous deposits are known as "brain sand". They increase in later life and may be a sign of degeneration in the organ. With the high power objective, note that the outlines of cells in the cords are not defined so that the cords show simply scattered nuclei in an indefinite ground substance. With special staining methods, neuroglial cells and "parenchymal" cells, both with cell processes, have been described. Pancreas (Slide 12) Study the islets of Langerhans under high power. The cells, with poorly defined outlines, are arranged in clumps and cords. The islets are abundantly supplied with capillaries, as befits an endocrine organ. The nuclei are roughly spherical with clearly defined particulate chromatin. In sections stained with H & E, the various cells look alike. With special stains, A, B, C and D cells have been differentiated depending on the staining properties of the granules. The B (or Beta) cells are responsible for insulin production and, for reasons yet unknown, occasionally limit their output thus producing diabetes mellitus.

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