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MODULE 15: MALE REPRODUCTIVE SYSTEM

WARNING: ALL THE MATERIALS (i.e., TEXT DOCUMENTS, DIGITAL
SLIDES, PHOTOMICROGRAPHS, PICTURES, VIDEOS, ETC.) THAT YOU WILL USE IN THIS LABORATORY COURSE IN HISTOLOGY ARE COPYRIGHTED. YOU ARE NOT ALLOWED TO DOWNLOAD, UPLOAD, SHARE, COPY, OR EDIT ANY PART OF ANY MATERIAL, UNLESS SPECIFICALLY GIVEN PERMISSION TO DO SO. YOU ARE HOWEVER ALLOWED TO TAKE PHOTOGRAPHS OF THE MATERIALS AS THEY APPEAR ON THE MONITOR, BUT ONLY FOR YOUR PERSONAL USE. YOU CANNOT POST THE PHOTOGRAPHS IN ANY SITE ON THE INTERNET, SEND THEM TO ANYONE BY E-MAIL OR ANY ELECTRONIC MEANS, OR MASS PRODUCE THEM. THIS LABORATORY IS MONITORED BY A CLASSROOM MANAGEMENT PROGRAM THAT CAN DETECT VIOLATORS OF THE ABOVE RULE. VIOLATORS WILL BE SUBJECTED TO DISCIPLINARY ACTION.
In males, the reproductive system consists of a pair of testes, the corresponding duct system of each testis, a copulatory organ (penis) and some accessory glands. The duct system of the testis can be grouped into intratesticular (i.e., within the testis) and extratesticular (i.e., outside the testis) The intratesticular genital ducts include the tubuli recti, rete testis, and ductuli efferentes while the extratesticular genital ducts consist of the ductus epididymis, ductus deferens (vas deferens), ejaculatory duct and urethra. The accessory glands of the male reproductive system, on the other hand, consist of the unpaired prostate gland, and the paired seminal vesicles and bulbourethral glands (glands of Cowper).

COMPETENCIES
At the end of the module on the Male Reproductive System, given an LM histologic section, the student can: 1. Name the organ where the histologic section was taken from. 2. Delimit the mediastinum testis. 3. Delineate the different regions of the testicular stroma: tunica albuginea and septulae testis 4. Delimit the testicular lobules. 5. Identify the seminiferous tubules. 6. Locate the gametes (in their various stages of development), Sertoli cells and interstitial cells (of Leydig) in the testis. 7. Distinguish the various spermatogenic cells from each other. 8. Identify the parts of the sperm cell. 9. Identify the histologic layers and distinctive features of the various intratesticular and extratesticular genital ducts. 10. Delimit the cavernous bodies that comprise the penis. 11. Point out the distinguishing features of erectile tissue. 12. Identify the histologic layers and characterisitic features of the accessory glands of the Male Reproductive System

PREREQUISITE KNOWLEDGE
Esteban and Gonzales Textbook of Histology, Fourth Edition Chapter XVI Male Reproductive System, pages 217 232

LABORATORY SESSION ON THE MALE REPRODUCTIVE SYSTEM

In this laboratory session, you will be studying digital slides only. You may have to consult your textbook and atlas in the course of the session. SLIDE SESSION Split the monitor and start/run Imagescope as you did during the previous lab sessions. Open the digital slides listed below. The slides are in the folder labeled Module 15. Male Reproductive System that is within the folder labeled HISTOLOGY SLIDES, which is on your desktop. Epididymis 1 Epididymis 2 Penis Prostate 1
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 TESTIS

Prostate 2 Prostate 3 Seminal vesicle Sperm Testis 1 Testis 2 Testis 3 Vas deferens 1 Vas deferens 2 Vas deferens 3

In studying the testis, you will use the following slides, which are already on the filmstrip: Testis 1 Testis 2 Testis 3

The testes are where spermatogenesis (i.e., the production of sperm cells or spermatozoa) occurs. They also produce the male hormone, testosterone. On the filmstrip, click the slide labeled Testis 1 to place the slide in the main window. The slide contains a sagittal section of a testis. o Study the section at low magnification. Delineate the connective tissue capsule, which is called tunica albuginea. Note that it is rather thick and is made up of dense irregular connective tissue. Locate the mediastinum testis. This is a region on the posterior surface of the testis (upper left area, in this slide) where the capsule is thickened and projects into the substance of the testis. It is at the mediastinum testis where blood and lymphatic vessels enter and/or exit, and the ductuli efferentes leave, the testis. Note that connective tissue elements from the mediastinum testis form septa (septulae testis) that invade and divide the organ into unequal pyramidal compartments called lobules (lobuli testis). Note further that the section shows quite a number of these lobules. Each testis has up to 250 lobules. o Study the lobules at higher magnification. Note that they are filled with tubular structuresshown mostly in cross and oblique sectionsthat are are separated
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from each other by a thin amount of connective tissue (actually, this is reticular tissue) where some cellular elements are embedded. The tubular structures are called seminiferous tubules. Seminiferous tubules are fine but long and highly-coiled tubes that are filled with fluid. Each testicular lobule contains 1-4 seminiferous tubules that start as blind or anastomosing tubes at the base of the lobule. They end on the apical portion of the lobule by becoming continuous with the tubuli recti. o Examine some of the seminiferous tubules at different magnifications. Note that their wall is formed externally by a thin fibrous sheath and internally by a stratified epithelium. The fibrous sheath contains contractile cells (myoid cells) that resemble smooth muscle cells. While the the epithelium consist of spermatogenic cells (i.e., gametes in various stages of development) and supporting cells called Sertoli cells. Find myoid cells. In the epithelium, locate Sertoli cells. Sertoli cells are large, tall cells. They have an elongated nucleus that is usually positioned at right angle to the basal membrane. The nucleus has a visible nucleolus and clumped chromatin material. Their broad bases rest on the basal lamina and their tapering apices extend into the lumen of the seminiferous tubule. On their lateral surfaces, they form folds where developing gametes are lodged. Near their bases, Sertoli cells form tight junctions with each other, which divide the lumen of the seminiferous tubule into a basal compartment and an adluminal compartment. Among the cells in the epithelium, identify typical examples of these spermatogenic cells: spermatogonia, primary spermatocytes, spermatids and spermatozoa. The spermatogenic cells, which are partially enveloped by the Sertoli cells, form 4-8 layers in the epithelium. The most undifferentiated cells (i.e., spermatogonia) occupy the basal compartment of the seminiferous tubule. As the developing gametes mature, they move towards the lumen of the seminiferous tubule. Spermatogonia are small cells (about 12 m in diameter). There are actually several types of spermatogonia, the main types are: dark type A spermatogonium (type Ad spermatogonium), pale type A
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spermatogonium (type Ap spermatogonium), and type B spermatogonium. The type Ad spermatogonium has a large ovoid nucleus that contains dense chromatin material that makes it appear relatively dark in routinely prepared histologic specimens. Type Ap spermatogonia look like type Ad spermatogonia except for their paler nucleus that usually contains a couple of nucleoli that are often associated with the nuclear membrane. Type B spermatogonia, on the other hand, resemble type Ap spermatogonia in appearance except that their nucleus is more rounded. Primary spermatocytes migrate to the adluminal compartment as soon as they have formed. They are easy to spot because they are the largest cells in the spermatogenic series (male germ cell lineage). They have abundant cytoplasm and a large nucleus whose chromatin material is in the form of fine threads or clumps. Secondary spermatocytes, which arise after primary spermatocytes complete meiosis I, are about half the size of primary spermatocytes. They are difficult to find in LM sections because they complete meiosis II to become spermatids rather quickly (i.e., in a matter of hours.) The spermatids are also easy to spot in LM preparations because aside from being the most numerous, they are also the smallest of the immature gametes. They are also the most internally situated among the immature cells in the seminiferous tubules. Spermatozoa are also easy to spot because they have an elongated nucleus and a flagellum or tail. o Now, focus your attention on the tissue that fills the spaces between the seminiferous tubules. Study it under different magnifications. Identify reticular cells. Identify interstitial cells (of Leydig). The tissue that is in between the seminiferous tubules consists of a highly vascular reticular tissue where reticular cells, mesenchymal cells, macrophages and interstitial cells (of Leydig) are embedded. The interstitial cells of Leydig occur singly or in clusters. They are also easy to find because they are large cells with copious cytoplasm that is is highly eosinophilic. The nucleus of the cells of Leydig is round. It contains fine chromatin and 1 to 2 peripherally located nucleoli. The interstitial cells (of Leydig) produce the male hormone, testosterone.
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o Shift your attention to the area of and around the mediastinum testes. Examine the area under different magnifications. Observe that the area consists of dense irregular connective tissue where numerous epithelial-lined spacesmost of which have irregular lumensare embedded. These epithelial-lined spaces are actually tubules and canals that are segments of either the tubuli recti or rete testis. o In the mediastinum testis, the seminiferous tubules continue into the tubuli recti, which are short, narrow and straight tubules that are lined by a simple epithelium. In its initial segment, the epithelium of the tubuli recti is made up of Sertoli cells that have no associated gametes. Distally, the epithelium consists of columnar or cuboidal cells that possess numerous microvilli. o The tubuli recti continue into the rete testis, which refers to a network of anastomosing canals that have irregular lumens, which are lined by simple squamous or simple cuboidal epithelium. Find tubuli recti (straight tubules). They are the tubes with regular lumens and lined by columnar or tall cuboidal epithelium that are near the seminiferous tubules. Locate tubuli recti that empty into the rete testis. Delimit the canals that comprise the rete testis. They have irregular lumens and their lining epithelium is simple squamous or simple cuboidal. They are more numerous in this specimen than the tubuli recti. On the filmstrip, click the slide labeled Testis 2 to place it in the main window. Reinforce what you have learned about the testis in the previous slide by studying this one using different magnifications. Click the slide labeled Testis 3 on the filmstrip to place it in the main window. Further re-inforce what you know about the testis by studying this one under different magnifications.

SPERMATOZOA
On the filmstrip, click the slide labeled Sperm to put the slide, which consists of a smear of male ejaculate in the main window. o Pan the slide at high magnification and note the numerous sperm cells (spermatozoa). o Look at some sperm cells more closely and identify the head and tail (flagellum). Note that the head consists of an elongated nucleus and an acrosome (acrosomal cap). In the tail, take note of the middle piece, which is the thickest segment of the tail. The tail of the spermatozoon actually consists of four segments: a neck, which attaches the tail to the head; a middle piece, which is thick because of its mitochondrial sheath; a principal piece, which is the longest part of the tail; 6

and, an end piece, which is the short distal part that is thinner than the principal piece.

EPIDIDYMIS, DUCTUS EPIDIDYMIS AND DUCTULI EFFERENTES You will need the following slides, which are already on the filmstrip in studying the epididymis, ductuli efferentes and ductus epididymis. Epididymis 1 Epididymis 2

On the filmstrip, click the slide labeled Epididymis 1 to put the slide in the main window. This slide contains cross-sections of the head of the epididymis and vas deferes. o Look at the thumbnail of the slide, note that there are essentially two specimens mounted on the slide. The smaller specimen on the right, which contains a single tubular structure is actually a section of the vas deferens. You need not study this section because there are better slides of the vas deferens that you will study later in this session. . o Focus your attention on the specimen on the left side of the slide, which consists of a section of the head of the epididymis. Study the slide under low magnification. The epididymis is s C-shaped structure that is attached to the posterior surface of the testis. The epididymis has 3 regions, which, from the most superior to the most inferior, consists of the head, body and tail. The epididymis is made up of connective tissue where blood vessels and segments of the duct system of the testis are embedded. In so far as the testicular ducts are concerned, the head of the epididymis contains the distal portions of the ductuli efferentes and the initial portion of the highly coiled ductus epididymis into which the ductuli efferentes drain; while the body and tail contains the rest of the ductus epididymis. Observe that the specimen consists of dense connective tissue where two types of tubular structures are embedded. These tubular structures are segments of the tubuli efferentes (efferent ductules) and the ductus epididymis. They are easy to tell apart because the ductuli efferentes have smaller lumens than the ductus epididymis. Furthermore, the lumens of the dutuli efferentes have irregular shapes. Direct your attention on the tubuli efferentes and examine them under different magnification. The ductuli efferentes consists of several fine and highly-coiled tubules that connect the rete testes to the ductus epididymis.
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The wall of the ductuli efferentes consists of an inner epithelium and an external thin layer of circularly arranged smooth muscle cells. The epithelium has a scalloped appearance in routine LM preparations because it is made up of groups of two types of cells (one type is columnar and ciliated while the other is cuboidal and nonciliated but contains microvilli) that line the duct in an alternating manner. Note the scalloped appearance of the epithelium. Verify that the epithelium is made up of two types of cells that form clusters that line the ductuli efferentes in alternating fashion. Try to distinguish the columnar ciliated cells from the cuboidal cells in the epithelium. Delineate the smooth muscle cells that form a layer external to the epithelium. Note the dense connective tissue that surround the ductuli efferentes. Now, shift your attention to the segments of the ductus epididymis in the specimen and study them using different magnifications. The ductus epididymis is a fine (diameter = 1mm), long (more than 6 meters) but highly coiled tube. Its wall, like that of the ductuli efferentes, consists of an epithelium that is surrounded externally by a thin layer of smooth muscle. But the epithelium is pseudostratified columnar that has a uniform rather than scalloped-shaped luminal surface. Two cell types comprise the epithelium of the ductus epididymis: principal cells and basal cells. The principal cells, which are more numerous, are provided with stereocilia. The basal cells, on the other hand, are small, rounded, or pyramidal cells that rest on the basal lamina in between the bases of the principal cells. Under the LM, they have a pale-staining cytoplasm and coarse nuclear material. Verify that the epithelium of the ductus epididymis is pseudostratified columnar. Distinguish the principal cells from the basal cells. Delimit the layer of smooth muscle cells external to the epithelium. Note the connective tissue that surround and bind together the coiled segments of the ductus epididymis.

On the filmstrip, click the slide labeled Epididymis 2 to place it in the main window. The slide contains a section of either the body or tail of the epididymis.

o Study the slide under low magnification. Note that specimen in the slide consists of numerous tubular structures that are bound together by connective tissue. These tubular structures are sections of the ductus epididymis. o Study the sections of the ductus epididymis under different magnifications. Note the sperm cells that occupy the lumen of many of the sections. Verify what you have learned about the wall of the ductus epididymis from the previous slide. In particular, note that the stereocilia of the principal cells are better defined in this slide than in the previous one. VAS DEFERENS (DUCTUS DEFERENS) In studying the vas deferens, you will need the following slides, which are already in the filmstrip: Vas deferens 1 Vas deferens 2 Vas deferens 3 On the filmstrip, click the slide labeled Vas deferens 1 to place it in the main window. The slide contains a cross-section of the vas deferens. o Locate the vas deferens (ductus deferens) under low magnification. This should be easy to do because the vas deferens is the big tubular structure with a very thick wall that occupies the central area of the slide. o Examine the vas deferens using different magnifications. Delineate the histologic layers that comprise the wall of the vas deferens: mucosa, muscularis and adventitia. Scrutinize the mucosa. Note that it forms folds that lends an irregular shape to the lumen of the vas deferens. Note further that it consists of an epithelium and a lamina propria. Observe that the epithelium is similar to that of the ductus epididymis in that it is pseudostratified columnar (with stereocilia) but the epithelium of the vas deferens is not as tall as that of the ductus epididymis. Notice that the lamina propria, which underlies the epithelium, is made up of loose connective tissue. Now focus your attention on the muscularis layer. Note that layer is very thick. Delineate the three layers that the smooth muscle fibers form: outer and inner layers where the cells are longitudinallyoriented, and a middle layer where the cells are circularlyoriented. Shift your attention to the adventitia.
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Notice that it is made up of loose connective tissue. Take note of the the blood vessels and nerves that are embedded in the connective tissue.

Go back to the filmstrip and click on the slide labeled Vas deferens 2 to place it in the main window. As in the previous slide, the slide contains a cross section of the the vas deferens. Re-inforce what you have learned about the histology of the vas deferens in the previous slide by studying this one using different magnifications. Now, click the slide labeled Vas deferens 3 to place the slide in the main window. The slide contains a section of the spermatic cord. The vas deferens, from its initial segment until it enters the abdominal cavity forms part of the spermatic cord. The spermatic cord consists of several structures that enter or leave the testis that are bound together by connective tissue. Aside from the ductus deferens and its blood vessels, the structures that comprise the spermatic cord include the testicular artery, pampiniform plexus of veins, and lymphatic vessels and nerves of the testes. o Pan the slide under low magnification and locate the vas deferens. It is in the upper left region of the slide. o Reinforce what you have learned about the histology of the vas deferens in the previous two slides by examining the vas deferens in this slide using different magnifications. o Now, focus your attention to the rest of the spermatic cord, which you should examine using different magnifications. o Note the numerous blood vessels and nerves. o Note the connective tissue that bind the structures together.

PENIS On the filmstrip, click the slide labeled Penis to place it in the main window. The slide contains a cross-section of the penis. o Look at the thumbnail of the slide. Note that the specimen mounted on the slide consists of three (3) compartments, that are separated from each other by thick layers of connective tissueone compartment is fully shown while only parts of the other two are shown in the slide. The three compartments are the paired corpora cavernosa penis, which refer to the two that occupy the right and upper regions of the slide and the unpaired corpus spongiosum penis, which occupies the region on the lower left half of the slide. The penis is mainly made up of three cylindrical masses (cavernous bodies): two corpora cavernosa penis, which lie side by side on the dorsal region of
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the penis and the corpus spongiosum peniswhere the spongy urethra (penile urethra; cavernous urethra) is embeddedthat occupy the midventral region of the organ. A dense connective tissue layer that is richly supplied with elastic fibers, the tunica albuginea, binds the three cavernous bodies together and forms a capsule around each one. Externally, the penis is covered by skin. The cavernous bodies are composed of erectile tissue (i.e., tissue that consists of numerous interconnecting endothelium-lined vascular channels that are separated from each other by connective tissue that contains elastic and smooth muscle fibers). In the presence of erotic stimuli, the vascular channels rapidly fill up with blood causing the penis to erect. o Pan and examine the corpora cavernosa under different magnifications. Observe the variation in shape and caliber of the lumen of the numerous vascular channels that comprise the two cavernous bodies. Note the presence of blood in the lumen of many of the venous sinuses. Delimit the endothelium that lines the sinuses. Note the presence of smooth muscle cells in the connective tissue that is in between the venous sinuses. Also note the presence of numerous thick-walled arteries and nerves in the erectile tissue of the corpora cavernosa. The arteries supply the vascular channels while the nerves provide the autonomic control that is needed in the process of penile erection. In the connective tissue external to the tunica albuginea that encloses the corpora cavernosa, note the presence of veins. o Now shift your attention to the corpus spongiosum and study it under different magnifications. Locate the spongy urethra. This should be easy because it occupies the center of the corpus spongiosun. Observe that the mucosa of the spongy urethra forms folds. Observe further that the mucosa consists of an epithelium and a lamina propria. Note that the epithelium is nonkeratinized stratified squamous. Most of the spongy urethra is really lined by pseudosteratified columnar or stratified columnar epithelium, but in certain areas and terminally the epithelium is nonkeratinized stratified squamous. Also note the presence of some small glands (mucus) and veins in the lamina propria. Notice the erectile tissue that is external to the lamina propria. Take note of the venous sinuses. Also note the occasional presence of arteries and nerves.
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PROSTATE GLAND In studying the Prostate gland, you will need the following slides, which are already in the filmstrip: Prostate 1 Prostate 2 Priostate 3

The prostate gland is the largest of the accessory glands of the male reproductive system. The urethra passes through it, but the sections you will study in this lab session do not include the prostatic urethra. The prostate is often referred to as a fibromuscular organ because its stroma which consists of dense irregular connective tissue is richly supplied with smooth muscle fibers. The stroma consists of a rather thick capsule that envelops the gland; septa from the capsule that penetrate the gland and divide it into ill-defined lobules and connective tissue and smooth elements that form the supporting framework for the alveoli within the lobules. Blood vessels and nerves are embedded in the stroma. The parenchyma of the prostate gland consists of the compound tubuloalveolar glands that occupy the lobules. The epithelium that lines the alveoli is highly variable. It is often simple columnar but in areas it can be simple squamous, simple cuboidal, or pseudostratified columnar. On the filmstrip, click the slide labeled Prostate 1 to place it in the main window. The specimen in the slide is a small section of the prostate gland. o View the section under low magnification. Delineate the capsule, which in this section is only partially seen. It is the thick connective tissue layer on the superior edge of the section. Note that the capsule gives off septa that invade the gland and divide it into ill-defined lobules. Observe that the lobules are occupied by glandular tissue. o Now, examine the capsule and the septa at higher magnifications. Note the numerous smooth muscle cells in the capsule and septa. On the left upper region of the slide notice the presence of clusters skeletal muscle cells that are in longitudinal and cross sections. These skeletal muscle cells are outside of, and are not part of, the prostate. Observe the presence of blood vessels and nerves in the capsule and septa. o Shift your attention to the alveoli that are in the lobules and examine them under different magnifications.

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Note that the alveoli are surrounded and separated from each other by connective tissue and smooth muscle elements that arise from the septa. Note that the lumen of the alveoli vary in caliber. Note further than the lumen of the alveoli also vary in shape because the lining epithelium and the underlying connective tissue form numerous irregular folds. Observe that the alveoli are lined by simple epithelium, which in this section consists mostly of low columnar cells and, in some areas, cuboidal cells.

On the filmstrip, click on the slide labeled Prostate 2 to place the slide in the main window. The slide contains a section of an adult prostate. o Reinforce what you have learned about the histology of the prostate in the previous slide by examining this one under different magnifications. o Observe that in the slide, the lining epithelium of the alveoli in the specimen is more variable than in the specimen in the previous slide. o Also noteworthy in this section is the presence of corpora amylacea (prostatic concretions) in the lumen of some of the alveoli. Corpora amylacea (prostatic concretions) are lamellated solid, sometimes calcified, structures that vary in shape and size. They are made up of glycoproteins and other substances that are deposited around cell fragments. They increase in size and number with age.

Go back to the filmstrip and click the slide labeled Prostate 3 to place the section in the main window. o Reinforce the knowledge you gained about the histology of the prostate from the previous two slides by studying this slide using different magnifications. o Note that the smooth muscle cells in the stroma of the prostate are easier to distinguish in this slide than in the previous two.

SEMINAL VESICLE On the filmstrip, click the slide labeled Seminal vesicle to place it in the main window. Males have a pair of seminal vesicles (left and right) that lie between the fundus of the urinary bladder and the rectum immediately above the prostate gland. o Look at the filmstrip of the seminal vesicle.Note that the slide exhibits the cross-section of several tubular structures that are bound together by loose connective tissue. These tubular structures are segments of a single tube because the seminal vesicle is a gland that is in the form of one long tube.
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A seminal vesicle consists of a single fine but very long tube (10-15 cm). The tube is highly coiled and the coils are bound together by connective tissue such that the seminal vesicle appears grossly as a saclike structure that is only about 5 cm long. Also, when you take a histologic section of the gland, you will invariably cut through several coils of the gland. o Examine the coils of the seminal vesicle using different magnifications. Delineate the three histologic layers of the seminal vesicle: mucosa, muscularis and adventitia. Focus your attention on the mucosa. Note that it forms numerous intricately branched folds that give the lumen the appearance of a maze. Note further that the mucosa is made-up of an epithelium that is supported by very fine connective tissue elements. Observe that the epithelium is nonciliated stratified columnar. Notice that the epithelium is populated by two types of cells: columnar, which comprise majority of the cells, and small, round cells that are located between the bases of the columnar cells. Now, shift your attention to the muscularis and delineate the two layers of smooth muscle cells that comprise the muscularis. This is difficult to do in this slide because the layers are not well defined. The muscularis consists of a thin inner layer of circularly arranged and a thicker outer layer of longitudinally arranged smooth muscle fibers. Direct your attention to the adventitia. Note that it is made up of loose connective tissue that is richly supplied with adipocytes, blood vessels and nerves. Note further that the adventitia serves to bind the coils of the seminal vesicle together.

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