Lab 9: Anatomy of the Digestive and Reproductive Systems Laboratory

The first part of this laboratory is intended to provide you with the basic anatomical features of the digestive system. There will also be a couple of histological sections that you will need to look at. The second part will introduce you to the major anatomical features of the reproductive system. You will want to bring your book for this lab. Next week we will review the major anatomical features of the cadaver and perform a reproductive physiology exercise. Introduction to Anatomy of the Digestive and Reproductive Systems: Most of the nutrients that we consume cannot be used in their existing state. Instead they must be broken down into smaller components, deposited into the bloodstream for transportation to the cells of the body. The digestive system has two anatomical subdivisions: the accessory organs and the digestive tract. The accessory organs are various structures in the body that directly aid in the breakdown of food into stuff the body can absorb. The accessory organs of the digestive tract include the teeth, tongue, salivary glands, liver, gallbladder and pancreas. The three primary organs (liver, pancreas and gall bladder) are not part of the tube within the body, but empty into the alimentary canal. The digestive tract is a tube extending from the mouth to anus and includes the oral cavity, pharynx, esophagus, stomach, small intestine and large intestine.

TORSO MODEL:
Organs of the Digestive Tract:
Most of the digestive tract (Saladin Fig 25.1) is located within the peritoneal cavity (Saladin A7A10; pg 36-40) of the abdomen. Along the dorsal wall of the abdomen, the parietal peritoneum turns inward and forms a sheet of tissue called the dorsal mesentery, extending to the digestive tract. This membrane then forms the serosa that outermost tunic layer, or outer covering of the stomach and most parts of the intestines. In some places it continues beyond the digestive organ as a sheet of tissue called the ventral mesentery, which may hang freely in the abdominal cavity or attached to the ventral abdominal wall. (You cannot see this on the models but they are nicely shown by Saladin Figure 25.3.) A long the lesser curvature of the stomach, the serosa of the anterior and posterior stomach surfaces meet and continue as a ventral mesentery called the lesser omentum and extends from the stomach to the liver (fig. 25.1). Along the greater curvature of the stomach, the serosae form the greater omentum, which hangs loosely over the small intestine like an apron. You cannot see either of these structures on the torso model otherwise the organs in the cavity would not be visible. At its inferior margin, the greater omentum turns back on itself, passes upward and forms serous membranes around the spleen and transverse colon. Beyond the transverse colon, it continues as a mesentery called the mesocolon which anchors the colon to the posterior abdominal wall. The importance of these membranes is to anchor the organs of the digestive tract within the abdominal cavity. Think about what it would feel like if the organs were not anchored, but instead were free floating. Would you ever want to jump up and down? The Oral Cavity: The oral cavity serves many functions including ingestion, mastication, initial chemical digestion, and swallowing. Define each of the terms: Ingestion: Mastication: Digestion The cheeks and lips retain food and push it between the teeth for mastication. Identify the upper and lower lips. Why are the lips fleshy and soft? The tongue is a muscular, bulky organ designed to manipulate food. Its surface is covered with stratified squamous epithelium and exhibits numerous bumps and projections called lingual papillae. Most of the lingual papillae are taste buds. Review the information in chapter 16 for what these receptors look like and their distribution across the surface of the tongue. The palate separates the oral cavity from the nasal cavity. Its anterior portion is the hard palate (or bony palate) and is a bony projection of the palatine processes of the maxillae and palatine bones (figure 9.4, page 255). Posterior to the hard palate is the soft palate which has a spongy texture and is composed mainly of skeletal muscle and glandular tissue. Clinically remember the discussion about the cleft palate?

The soft palate has a conical medial projection called the uvula -- the piece of tissue that hangs down in the back of the mouth. Try talking without letting your tongue touch the hard palate, if you had Fetal Alcohol syndrome or a congenital birth defect you might have been born without a hard palate or have some cleft palate defect. Why is the hard palate so important for speech? An adult normally has 16 teeth in the mandible and 16 in the maxilla (32 total). Collectively they are called the dentition. Review Saladin Fig. 25.5 and identify all tooth types: 8 incisors, 4 canines, 8 premolars and 12 molars, also be able to identify these on the teeth of a actual human skull. What is the function of each tooth type? How might dental caries negatively impact your ability to eat and digest food? The salivary glands (Saladin Figure 25.8) can be divided into two broad groupings called the intrinsic and extrinsic salivary glands. The intrinsic glands are located within the oral tissue. These glands secrete relatively small amounts of saliva at a fairly constant rate, keeping the mouth moist and inhibiting bacterial growth. They also produce lingual lipase and lysozyme. Lingual lipase is an enzyme that begins to break down lipids (i.e., fats). Lysozyme is an enzyme that has some antibiotic properties. The extrinsic glands are the more obvious glands, and the ones you can see on the model. These glands are situated outside the oral cavity. Locate the parotid gland, submandibular gland and sublingual gland on the torso model. The parotid glands are just ventral to the ears and cover the maxillarymandibular joint. The submandibular glands are typically close to the medial side of the mandibles and the sublingual gland is directly below the tongue. When your doctor gently palpates under your jaw, he/she is attempting to tell if your glands are swollen. If you have an infection, these glands may be enlarged. The best example of such swelling is with the mumps where the parotid glands enlarge. The pharynx is a common passageway for the respiratory and digestive system. It extends from the dorsal extensions of the nasal cavity, past the posterior aspect of the oral cavity and down towards the larynx. Identify this organ on the torso model. It is typically divided into the nasopharynx, which includes the area just behind the nasal cavity; the oropharynx, the area behind the oral cavity; and the laryngopharynx, just anterior to the larynx. It takes about 2 seconds for you to swallow food from the oropharynx to the stomach. The esophagus is a straight muscular tube that begins posterior to the larynx at the level of the cricoid laryngeal cartilage and travels downward through the mediastinum of the thorax. This is the major tube that allows food to enter into the stomach. The esophagus penetrates the diaphragm at an opening called the esophageal hiatus and meets the stomach at the cardiac orifice. This orifice is surrounded by smooth muscle called the lower esophageal sphincter. Locate the esophagus and its regions in the torso model. Note that it runs dorsal to the trachea. You have actually seen this on the male cadaver as it extends down the thoracic cavity towards the diaphragm. The wall of the esophagus is organized into the following tissue layers: Mucosa, Submucosa, Muscularis Externa and Adventitia/Serosa. All organs have these tissue layers in very similar patterns. Anatomically this is a unique structure because the esophagus starts as a tube of skeletal muscle and makes a transition to all smooth muscle at the cardiac orifice. If you had gastroesophageal reflux disease, where would you see the lesions? What contributes to the lesions? When a pill is not swallowed properly with adequate water, it may get stuck in the esophagus and forms an adhesion to the both sides of the esophagus. The pill dissolves and injures the underlying tissues giving it a kiss-like appearance. Do a quick search on line for kissing ulcers (nothing funky here ok?). The stomach is a muscular sac in the upper abdominal cavity immediately inferior to the diaphragm. It is slightly J-shaped. The medial margin is called the lesser curvature and the longer lateral surface is called the greater curvature. The stomach is divided into four regions: the cardiac region, the fundic region, the body, and the antrum. Each region has a particular function and secretion. Based on figure 25.11, identify each of the four regions of the stomach on the torso model. Draw a picture of the stomach and label the four regions. When you have gas, what part of the stomach floats and causes the discomfort? Why do they call gastric gas heart burn sometimes? If you had an ulcerative lesion that eroded through the stomach wall to the peritoneal cavity, what would the outcome be?

These types of Gastric Bypass Surgery have become popular for weight loss. Here are some different types that are discussed at http://www.nlm.nih.gov/medlineplus/ency/article/007199.htm

Roux-en-Y stomach surgery for weight loss

Adjustable gastric banding

Vertical Biliopancreatic Biliopancreatic Dumping banded diversion diversion with syndrome gastroplasty (BPD) duodenal switch

Nearly all chemical digestion and nutrient absorption occurs in the small intestine. The small intestine is divided into three regions: duodenum (about the first 8-10 inches), jejunum (the middle segment and is about 2.5 meters long), and the ileum (the last 3.6 meter segment). The duodenum begins pyloric sphincter and runs for The first 12 fingers widths of the intestine. The jejunum is fairly non-descript, but is the middle segment of the intestine. The jejunum and ileum are not separated by any conspicuous landmark, but the jejunum is typically located in the upper left of the intestinal coils and the ileum to the lower right. Most nutrient digestion and absorption occurs in the jejunum and ileum. The ileum ends at the ileocecal junction, where it joins the caecum, the first part of the large intestine. The large intestine begins at the cecum, a blind pouch in the lower right abdominal quadrant. (Saladin Figure 25.30) Attached to its lower end is the vermiform appendix. The large intestine is subdivided into the ascending colon, the transverse colon and the descending colon. The ascending colon begins at the ileocecal valve and passes up the right side of the abdominal cavity. It makes a 90 degree turn at the hepatic flexure (bend close to the liver) and then becomes the transverse colon. The transverse colon passes horizontally across the upper abdominal cavity and turns downward at the splenic flexure (bend near the spleen). It now becomes the descending colon which passes down the left side of the abdominal cavity. The pelvic cavity is narrower than the adbominal cavity so at the pelvic inlet the colon turns medially and downward, forming a roughly S-shaped portion called the sigmoid colon. In the pelvic cavity, the large intestine straightens and forms the rectum. The final 3 cm of large intestine is the anal canal. The muscularis externa of the colon is unusual in that its longitudinal fibers do not encircle the colon. Instead this muscle layer is divided into three ribbon-like strips called teniae coli. These small strips constrict and create little pouches called haustra. Locate these on the torso models. The colon has the unique distinction of having the greatest number of bacteria of any section of the digestive system, this can be good or bad. The good, some of these bacteria actually create things we need like vitamin K. The bad, some of these bacteria take undigested carbohydrates in things such as beans or broccoli and produce gas (flatus). Bacteria in this region can also produce toxic materials that can be carcinogenic to the epithelial cells that line the colon. Why is colo-rectal cancer relatively common? Why is it that diets rich in fiber (undigested fibers attract water to make the stools large and soft) help protect against colo-rectal cancer? In contrast diets rich in meat (have you ever smelled rancid meat?) are often associated with increased risk of colo-rectal cancer? During your colo-rectal exam, your proctologist will look for either obvious cancerous growths or precancerous growths called polyps. If you have parents who are older, they may be willing to share what this examination entails (ask them if you like). If you had colo-rectal cancer and had a portion of your large intestine removed, how would your digestive process change? How would this alteration affect bile salt recycling and production of nutrients by the microflora? What if you have Irritable Bowel Syndrome or Crohns Disease, would your reabsorption of digestive secretions be normal? What would be the effect?

Accessory Organs of the Digestive System:

Several of the accessory organs have already been discussed when examining the oral cavity. However the functions of the intestine rely on the actions of the larger accessory organs, in particular the liver and pancreas. Liver: The liver is the largest gland in the body. It is typically a reddish brown organ located immediately inferior to the diaphragm and fills most of the right hypochondriac and epigastric regions of the abdominal

cavity. The liver has a diverse number of functions including the secretion of bile that aids in fat emulsification and digestion. The liver also has an unusual ability to regenerate if a section is removed, an important thing for transplant donation. Please dont try this with your heart or lung! Check out this website: http://home.comcast.net/~wnor/liver.htm The liver has four lobes called the right, left, quadrate, and caudate lobes. The right lobe is the largest and the caudate lobe is the smallest. Using Figure 25.17, page 907, locate each of the lobes of the liver on the torso models. Note that the right and left lobes are separated from each other by the falciform ligament. In the inferior aspect, locate the quadrate and caudate lobes. The quadrate lobe is next to the gallbladder and the caudate is located next to the vena cava. Locate the gallbladder on the inferior surface of the liver. You should also be able to identify the hepatic portal vein, the hepatic artery and the common hepatic duct. What is a portal circulatory structure? Compare and contrast the degree of blood oxygenation in each of these three blood vessel types. Gallbladder: The gallbladder is a small greenish sac semi-embedded in the inferior surface of the liver. The head of the gallbladder projects lightly beyond the liver and is more balloon-like. The neck of the gallbladder is continuous with the cystic duct that joins with the hepatic bile duct to become the common bile duct. The gallbladder is primarily a holding chamber for temporarily holding the bile produced by the liver. When you digest foods the gall bladder contracts to expel the stored bile into the intestine. Locate each of the above structures on the torso models. Where do gall stones form and how do they create inflammation in the wall of the gall bladder and sometimes jaundice? Pancreas: The pancreas is a soft, spongy, pink gland that is posterior to the greater curvature of the stomach and outside the peritoneal cavity. Remember that the pancreas produces exocrine secretions that are critical for digestion. What ducts permit the exocrine secretions of the pancreas to be drained? The pancreas also produces endocrine secretions such as insulin and glucagon. What do these two hormones do? Why is blockage of a pancreatic drainage a cause for pancreatitis?

Histology of the Digestive System

Microscopic anatomy of the Esophagus/stomach transition (slide 28 from the drawer) This is a longitudinal section of the esophagus-stomach junction. So the first thing you need to orient yourself to what you are looking at. Locate the mucosa layer (the surface next to the lumen). The esophageal mucosa consists of stratified squamous epithelium. Nearer to the stomach junction there is a distinct muscularis mucosa layer. Both the mucosa and submucosa form longitudinal folds that give the lumen an irregular outline. The submucosa is filled with numerous glands that produce mucus to lubricate the surface of the epithelium. The submucosa also contains numerous small blood vessels. The muscularis externa layer in the esophagus transitions from skeletal muscle in the first 1/3, to a mix of skeletal and smooth in the middle third and finally to simply smooth muscle in the distal 1/3 of the organ. You should be able to identify two smooth muscle layers in the muscularis externa, an inner circular smooth muscle layer and an external longitudinal layer. At the gastro-esophageal junction the epithelium transitions from stratified squamous (esophagus) to simple columnar (stomach). The inner surface of the stomach is organized into longitudinal folds called rugae. Extensions of the mucosa and submucosa extend into the folds. The muscularis externa layer has three smooth muscle layers, a circular layer, transverse and longitudinal layer. This allows the stomach to have a wringing motion, the kind of motion one does with a wash-cloth to get rid of the water. Compare slide 24 (Fundic Region of the Stomach) with this portion of slide 28. Microscopic anatomy of the small intestine: (Slide number 5 from the drawer) The microscopic anatomy of the small intestine includes modifications for maximizing digestion and absorption, as well as glands and muscles to facilitate in this process (Saladin Figure 25.24). If you were able to examine the surface of the mucosa, you would see that the surface is covered by finger-like projections called villi. These function to increase the surface area of the mucosa. Each villus is covered by simple columnar epithelial tissue (absorptive cells) with numerous goblet cells interspersed in the layer. One the very surface of the epithelial cells are microvilli -- you probably cannot see these with your microscopes. These microvilli have many proteins embedded in their plasma membrane that function as digestive enzymes called the brush border enzymes. The villus is filled with areolar connective tissue of the lamina propria. Embedded in this areolar connective tissue is an arteriole, capillary network and a venule, as well as a lacteal -- a lymphatic capillary. Between the bases of many of the villi are intestinal crypts. These crypts are similar to the gastric glands of the stomach. The submucosa is characterized by numerous, predominant duodenal glands. Look for the semicircular structures that appear to be made of cuboidal epithelial tissue. These glands are important in secreting bicarbonate-rich mucus. This mucus will neutralize stomach acid and make the chyme ready for the digestive enzymes. The muscularis externa consists of a relatively thick inner circular layer and a thinner longitudinal layer. Draw a picture of the cross-section of the small intestine. Label the epithelial tissue, submucosa, muscular externa (both the longitudinal and circular layers), and serosa. What would happen to your digestive physiology the duodenum, ileum or jejunum were removed? Microscopic anatomy of the colon (large intestine) (slide 32 from drawer): Again your first task is to look at this slide on very low power and familiarize yourself with the tunic layers. The epithelial layer (simple columnar E. T.) lines the lumen. Together with the lamina propria and the muscularis mucosa (a very small smooth muscle layer that follows just internal to the E.T.) make up the mucosa layer. The submucosa is filled with connective tissue that extends into the villi of the colon. Finally, the muscularis externa layer has two smooth muscle bands, the inner one is circular and the outer, much reduced in size and distribution is the longitudinal layer. Microscopic anatomy of a salivary gland (slide number 21 in drawer): This slide is a composite of three tissue slices. Scan the slide and see if you can determine differences in the sections. Isolate the middle, mixed gland slice, this is more than likely a section of the submandibular gland. There are both musous cells and acinar cells. One one side you should see a gland with numerous mucous cells in fact the mucous cells make up most of the tissue prep creating very light almost cloud like cells. This is similar to the histology of the sublingual gland (Fig.25.10, p.963). The other tissue slice with predominantly serous acinus is the parotid gland. It is made up almost exclusively of acinar cells. You should be able to isolate mucous cells, serous cells, C.T. stroma, and ducts.

Microscopic view of the Liver: (You will use slide 23 for this exercise.) The liver parenchyma consists mostly of cuboidal cells called hepatocytes. These hepatocytes are arranged in approximately 6-sided (hexagonal) cylinders called hepatic lobules (Saladin Fig 25.20). Scan the microscope slide at 4 power (low power) to get an appreciation for the slide. Each lobule has a central vein passing through its core. This is not a true vein as in the vascular system, but rather as a central blood-filled canal. The hepatic lobules are separated from each other by a sparse connective tissue stroma. Locate a hepatic lobule on the slide. Where multiple lobules come together you may find a hepatic triad. This structure consists of two blood vessels and a bile ductule. Try and locate a triad. I will not ask you to identify individual vessels in the triad. The hepatocytes form thin epithelial plates that fan out away from the central vein, like the spokes of a wheel radiating from the hub of the tire. These plates are separated from each other by blood-filled channels called hepatic sinusoids. This means that the hepatocytes are in direct contact with blood. The blood vessels of the hepatic triad supply blood to the sinusoids and the blood filters through the sinusoids around the hepatocytes to the central vein. From the central vein blood ultimately flows into the right and left hepatic veins and leaves the liver. Sometimes the liver turns harmless things into potent toxins by accident, this can happen if you mix alcohol with acetaminophen(Tylenol), which can result in massive acute liver failure (this is a major cause of liver failure and need for a liver transplant). Why might the cells surrounding the sinusoid just before it enters the central vein by most likely to die form the acetaminophen metabolites? The liver secretes bile into narrow channels called bile cannaliculi that are found between the sheets of hepatocytes. The bile then passes into the hepatic ducts that ultimately lead to the common hepatic duct. You cannot see these structures under the microscope except for the bile ductule in the triad. What is jaundice? What happens to liver function when you have jaundice as an adult? Why are infants born with a trace of jaundice? Why does sunlight seem to help clear up jaundice in infants? Microscopic view of the pancreas: (you will review slide 20 for this exercise.)

You have actually looked at this slide before. Recall finding the islets of Langerhans. Now you want to focus on the exocrine portion of the pancreas. This is actually the bulk of the cells (98%) in the pancreas, excluding the islets. These cells serve an exocrine function, producing enzymes secreted in the pancreatic juices.

Structures of the Digestive System one should know for next exam
tongue canines sublingual gland stomach body of stomach jejunum ascending colon sigmoid colon left lobe of liver hepatic bile duct haustra of L.I. soft palate premolars nasopharynx lesser curvature of stomach antrum of stomach ileum hepatic flexure rectum quadrate lobe of liver cystic duct hard palate molars oropharynx greater curvature of stomach pyloric region of stomach ileocecal junction transverse colon anal canal caudate lobe of liver common bile duct uvula parotid gland laryngopharynx cardiac region of stomach pyloric sphincter caecum splenic flexure liver hepatic portal vein (associated with liver) gallbladder incisor submandibular gland esophagus fundic region of stomach duodenum vermiform appendix descending colon right lobe of liver hepatic artery (associated with liver) pancreas

Histology for next Exam

Know the epithelial tissues of all of the organs examined. Know the tunic layers and what structures delimit each layer. Know the specific smooth muscle layers in the muscular externa. Know the cell types when identified with the slide (e.g., salivary glands) With the S.I., be able to identify structures like the villi. You should be able to recognize the esophagus from the longitudinal section of the esophagus-stomach. Likewise you should be able to recognize the stomach from this same slide. You should be able to identify the specific salivary gland from the tissue description. You should be able to recognize the liver from the slide and the pancreas from the slide. At the liver, you should be able to recognize the vessels and ducts from the slide.

REPRODUCTIVE SYSTEM MODELS:

MALE REPRODUCTIVE SYSTEM MODEL: We will study the parts of this model in a sequence that follows the pathway of sperm through the male reproductive system. We will begin with the site of sperm production, the testis in the scrotum. The scrotum is a skin-covered, muscular sac that contains the testes. A part of the scrotum is shown on each half of the model. On the right half of the model, a part of the medial partition between the testes is shown. Within the scrotum, find the testes. The testis on the left side of the model is shown with its surrounding membranes removed. The epididymis can be found superior and medial to the testis on the same side of the model. It is held tightly against the testis by a complicated set of membranes that surround both organs. The pink ductus deferens can be seen attached to the inferior, medial tail of the epididymis. Next, find the spermatic cord. It is a complicated structure that contains the ductus deferens, a network of veins and arteries and a nerve. All of these structures are bound together by the same set of membranes that tie the testis and epididymis together. The spermatic cord continues upward to the abdominal wall where it penetrates the inguinal canal, a reinforced opening through the lower abdominal wall. On the left side of the model parts of the abdominal wall are removed so that the passage of the spermatic cord, through the inguinal canal, can be followed. The sperm-transporting structure within the spermatic cord is also the ductus deferens. This tan muscular tube can be traced from its origin on the epididymis, through the spermatic cord, through the inguinal canal and into the abdominal cavity. From the inguinal canal, trace it across the superior surface of the urinary bladder, over the ureter and down the posterior surface of the bladder to where it joins with the seminal vesicle and terminates in the prostate gland. Now split the model in half and remove the left side of the bladder and attached structures. The seminal vesicles can be located on the posterior surface of the urinary bladder. Each seminal vesicle is a multi-lobed gland, which is painted tan. Notice that their ducts join with the ductus deferens. Inferior to the seminal vesicle, find the prostate gland. This organ is positioned close to the bottom of the urinary bladder. The prostate gland is painted a dark red-brown. Note that the urethra passes through it. The short ejaculatory duct does not show up well on this model. It originates where the seminal vesicle and ductus deferens merge. This duct occurs entirely within the prostate gland. Its termination in the urethra is shown inside the prostate gland. Now, let us find the three parts of the urethra. The prostatic urethra is easy. It is the part of the urethra inside the prostate gland. Next, comes the membranous urethra. This term refers to that short segment of the urethra between the prostate gland and the base of the penis. On this model, it is painted a mottled yellow/orange. It is very short, approximately one-half inch long. Finally, find the spongy urethra, that segment of the urethra inside the penis. Examine a half of the entire model so that you are aware of the position of the base of the penis near the prostate gland. The spongy urethra traverses the entire penis through the blue-painted spongy column. The penis originates with muscular attachments near the prostate gland. Note that it is firmly anchored to the pelvis by ligaments. The expanded distal end of the penis is named the glans of the penis. To examine the penis in cross section put the two detachable pieces together. Three columns of the blue and red erectile tissue are visible. The two superior columns are named the cavernous columns. The inferior one, containing the urethra, is named the spongy column. Split the penis model in half to see that the spongy column expands to form the glans. A good exercise, at this time, would be to trace the pathway of sperm from where it is produced in the testes to where it exits through the penis. One possible difficulty you might encounter is the role of the seminal vesicle. Sperm never pass through or enter this gland. It merely produces a mucus secretion which contributes to the formation of semen. On the other hand, sperm cells do pass through the prostate gland. Can you figure out why that last statement is true? FEMALE REPRODUCTIVE SYSTEM MODEL: Use the independent plastic model (not torso model) for this section. Start on a sagittal section view of one-half of the model. Place the inner piece in the correct position inside the outer piece. Locate the urinary bladder, urethra and urethral opening. Next, find the uterus, vagina and vaginal opening. Finally, locate the rectum and anal opening.

With that orientation in mind, remove the inner piece from the true pelvis and combine it with the other half model to study the uterus, ovary and uterine tube. Let us begin laterally at the yellow-painted ovary. Two tubular structures pass from the ovary to the uterus. The pale pink one is a ligament that helps support the ovary. The dark pink/red one, with the red expanded end, is the uterine tube. The greatly expanded, funnel-like end of the uterine tube, which partly surrounds the ovary, is called the infundibulum. Finger-like fimbrae can be seen attached to the edge of the infundibulum. Next, consider the uterus which consists of three distinguishable parts. Superior to the insertion of the uterine tubes is the fundus of the uterus. The middle part is called the body of the uterus. To find the cervix of the uterus split the model in half. The cervix of the uterus is the narrower part which protrudes into the vagina. The recessed grooves between the cervix and vaginal wall (like a moat around a castle) are called fornices. Find the position of the anterior fornix and the posterior fornix. Internally, note that the uterus shows two different textures and colors. The inner striated zone, which is a darker red, represents the endometrium. External to the endometrium is a lighter red layer called the myometrium. The smooth, pink,outermost layer of the uterus is its serosa, called that because it is a serous membrane. Between the uterus and urinary bladder is a peritoneum-lined pouch called the vesicouterine pouch. (This pouch is not open on the model.) Between the uterus and rectum, find the rectouterine pouch. These two pouches are of medical significance because they are "low points" for the drainage of abdominal cavity infections. Note that the rectouterine pouch can be easily drained through the vagina by means of an incision in the posterior fornix. Now, reassemble the model so that we can examine the external genitalia. Note that both the urethra and vagina open into a narrow space called the vestibule. Lateral to the vestibule are two folds of skin. The inner, smaller folds are named the labia minora. Lateral to them are thicker folds called the labia majora. At the anterior part of the vestibule, note a blue-painted structure anchored to the pubic bones. This structure is the clitoris. Near the opening of the vagina, a small, horizontal fold of mucous membrane represents the hymen. OVARY MODEL: This model represents a half ovary with an assembly of the stages of development of follicles. At the middle bottom is a mature follicle which is in the process of ovulation: rupturing to shed its ovum. The ovum is the yellow structure being extruded. To the left of the mature follicle is a large, yellow, scalloped structure, a corpus luteum. It formed from a previously ovulated mature follicle. Between the mature follicle and corpus luteum is one of three corpora albicans. These are the remnant scars of previous copora lutea. To the left of the mature follicle are three medium-sized, immature, secondary follicles that may someday become mature follicles. REPODUCTIVE HISTOLOGY: Reproductive Endocrine Histology: Ovary slide (slide up at front of lab) (See Saladin Fig. 17.12 for drawing and Fig 28.14 for histology). What is the main hormone produced by the granulosa cells of the ovarian follicle? (This hormone belongs to the estrogen group of hormones.) _________________________ . If you look at the graph associated with the physiology instructions you will see that this hormone is the dominant one during the follicular phase and is responsible for causing the LH surge. This is a great website with some nice histology images.

http://instruction.cvhs.okstate.edu/histology/HistologyReference/HRFemaleRS.htm
What hormone is produced by the corpus luteum? ____________________________ This is the hormone of pregnancy. Human chorionic gonadotropin (hCG) from the pregnancy test physiology exercise is responsible for keeping these endocrine structures functional during the first weeks of pregnancy while the placenta is growing. Later the placenta will secrete most of the hormones associated with pregnancy. The home pregnancy test we will see next week uses hCG as the basis for determining if a person is pregnant. Draw a picture of a whole ovary. Show small and large ovarian follicles. Draw a circle around one and label it. Label granulosa cells and an ovum. You will not see any corpus luteum. Testis slide (slide # 12) (Saladin Fig. 17.12) The endocrine cells you need to focus on are between the seminiferous tubules. They are called interstitial cells. What hormone do these cells produce? _______________________________ Draw a picture of a section through the testis. Draw a circle around and label a seminiferous tubule. Also label developing sperm cells and interstitial cells.

Checklist for Reproductive models: Male model Scrotum Testis Epididymis ductus deferens spermatic cord inguinal canal seminal vesicle prostate gland ejaculatory duct prostatic urethra membranous urethra spongy urethra glans penis corpus cavernosum corpus spongiosum Urethra urethral opening Uterus Vagina vaginal opening Ovary round ligament of uterus uterine tube infundibulum of uterine tube Fimbrae fundus of uterus body of uterus cervix of uterus anterior fornix posterior fornix Endometrium Myometrium Perimetrium vesicouterine pouch rectouterine pouch Vestibule labium minus labium majus Clitoris Hymen-Female model Ovary model mature follicle Ovum corpus luteum corpus albicans secondary follicle

PREGNANCY TESTS AND HOW THE MOTHERS BODY KNOWS A FETUS IS PRESENT. Endocrine glands are dispersed throughout the human body and carry out diverse actions from the regulation of blood glucose levels by the pancreas to controlling reproduction and pregnancy by the pituitary and gonads. Hormones really do participate in all aspects of human physiology. Today our focus will be on hormonal participation in reproduction and will specifically address how a womans body is signaled when a developing embryo is present. We will all become familiar with the cellular mechanism of actions of the hormones from lecture, but these mechanisms are hard to associate with our daily activities. So instead, this laboratory exercise will focus on home pregnancy tests and how hormones in the woman's body change with pregnancy. Many of you may have watched old television shows where the wife whispers on the phone "The rabbit died." The husband instantly knew this encrypted message meant that he was going to be a father sometime within the next 8 or so months. This euphemism originated sometime in the 1920's--1930's to mean a positive pregnancy test. What was the origin of this saying? Urine from a pregnant woman, injected into a female rabbit was found to cause ovarian hyperplasia (i.e., corpora hemorrhagica), proliferation of ovarian tissue detectable under histological examination. Although all rabbits used for pregnancy determination died from this procedure, animals that were exposed to the compounds in a pregnant woman's urine were the only ones that had bulging hemorrhages on the ovaries. The euphemism became a way of sharing confidential information without being overly specific about the nature of the information -- remember it implied someone had sexual intercourse at sometime and given the 1920s - 1930s, this was a taboo subject. Estrogen is the female hormone associated with development of secondary sex characteristics and mate attraction. In particular, but also in very simplified terms, estrogens are the hormones that get the body ready to be impregnated and advertise this availability. In order for conception to occur, ovulation must take place and be synchronized with intercourse. Humans are not reflex ovulators that is to say women do not ovulate when they have intercourse. Women do ovulate spontaneously when endogenous hormones signal the ovum to be released from a Graafian follicle, during the ovarian cycle. The Ovarian Cycle: The ovarian cycle is complicated by the facts that all of the ova are initially made in the second trimester fetus and that once a primordial follicle is activated, it may take up to a year for a it to develop into a mature Graafian follicle that is capable of ovulation. Some ova may wait for up to fifty years before they reawaken to be included in an ovarian cycle. However, after adolescence and before menopause, there are ovarian cycles of about 28 days duration which function

R eview of the uterine/menstrual cycle. (This image was accessed Jan. 10, 2004 and copied from: http://www.people.virginia.edu/~rjh9u/menscyc3.h tm). SEE ALSO FIGURE 28.14, p. 1094 OF SALADIN.) to prepare the uterus for potentially welcoming the product of an ovulated egg which may have become fertilized in the uterine tube and then have developed into an early embryo there. About 6 days after fertilization, the embryo hatches from its zona pellucida and then implants into the endometrium of the uterine wall (note the development of the endometrium in the 5th panel of diagram).

The first day of the ovarian cycle coincides with the first day of the menstrual cycle and is marked by the onset of menstruation. At this time the ovaries contain a small number of tertiary follicles, some atretic follicles and several thousand smaller follicles. By day 3 some of the tertiary follicles have enlarged as FSH and LH levels rise. As these tertiary follicles grow they secrete estradiol (the most potent estrogen) and that hormone level rises. This marks the beginning of the proliferative phase of the uterine/menstrual cycle. By day 13, only 1 large follicle (maybe two) persists and is called a Graafian follicle, while the other large follicles degrade (undergo atresia). The rise in estradiol from the maturing follicles creates a positive feedback mechanism to the hypothalamus and promotes enhanced GnRH secretion (this is a long-loop feedback mechanism). Estradiol levels peak about day 13 and promote an LH surge. This LH surge facilitates ovulation in about 9-12 hours. Ovulation marks the end of the follicular phase. It is also the time in the menstrual cycle when a woman is most fertile. The wall of the ovulated follicle differentiates into a growing corpus luteum (CL) and this structure begins to secrete progesterone and some estrogens. Elevated progesterone levels act synergistically with the high estrogen levels to provide negative feedback to the hypothalamus which in turn shuts off GnRH secretion and suppresses LH and FSH. (This feedback mechanism suppresses follicle development -- and if you think about it, you really do not need a follicle maturing when there is a potential pregnancy.) At the same time, this estrogen and progesterone affects the development of the uterine endometrium. The Uterine/Menstrual Cycle: Each ovarian cycle is correlated with a uterine/menstrual cycle which is divided into three phases: 1) the menstrual phase (correlated with reduced progesterone and reduced estrogen production by the ovary), 2) the proliferative phase (correlated with the follicular/preovulatory phase of the ovary, during which estrogen is being produced), and 3) the secretory phase (correlated with the luteal/postovulatory phase of the ovary, during which estrogen and much progesterone is being produced). The first day of menstruation is designated as the first day of the uterine/menstrual cycle and the beginning of the menstrual phase. During this phase, part of the lining of the uterus degenerates and begins sloughing away. This is removing old tissue in preparation for the growth of new materials (panel 5). As illustrated in panel 2 and 4 of the included diagram, day 1 is when all hormone levels are low.

The proliferative phase begins on about day 7. The uterine endometrium begins to thicken and becomes vascularized under the influence of increasing estrogen levels. The endometrium grows from about 4mm thickness to about 14mm thick. The secretory phase of the uterine/menstrual cycle begins on about day 14/15, immediately after ovulation. It is really all about thickening the endometrium, development of uterine glands, and preparing the uterus for pregnancy. This development is stimulated by estrogen and progesterone from the corpus luteum. The uterine glands secrete substances thought to be important during the first trimester of pregnancy for conceptus implantation, fetal nutrition, and development of the placenta. From Ovulation to Conception: Usually ovulation occurs 13 to 15 days after the first day of the last menstruation. Once ovulated, the egg remains viable only about 6-24 hours. Sperm is viable about 28-48 hours. Fertilization of the egg typically happens in the uterine tube. From there the conceptus travels to the uterus and implants in the uterine wall. Fetal and maternal tissues jointly make the placenta. It will take several weeks for the placenta to fully form. Back at the ovary, the corpus luteum continues to produce progesterone to sustain the uterus. Progesterone is the hormone responsible for maintaining pregnancy in the woman. Without adequate progesterone levels present, pregnancy will be terminated spontaneously. However, it takes a short while for the cells of the corpus luteum to respond, differentiate, and become progesterone-sectreting machines, given that a pregnancy ensues. It also takes several weeks for the placenta to develop, which then secretes progesterone and replaces this function of a corpus luteum. (Remember, not every ovulation results in a pregnancy.) So how do these endocrine-secreting machines -- the corpus luteum-- know a pregnancy has occurred? What is the simplest way to signal the female's body that she has conceived? (Law of parsimony.) The best way to determine if conception has occurred is for the woman's body to recognize a signal directly from the fetus and/or fetal tissues. Human chorionic gonadotropin (hCG) is that chemical hormone signal. The trophoblast cells of the early embryo and their derivatives in the placenta (a collection of fetal and maternal cells) secrete hCG. Measuring the presence of this hormone is also a good way for us to determine if someone may be pregnant. Human chorionic gonadotropin is a trophic hormone and as such

stimulates the corpora luteum to differentiate and grow, ultimately augmenting the progesterones and estrogens the ovary normally produces. This facilitates maintaining a uterine environment suitable for pregnancy. Let's take apart the name -- human chorionic gonadotropin. We are dealing with humans, so that takes care of that designation. This is a gonadotropin. Tropic hormones are hormones that cause the release of other hormones from a distant endocrine tissue. The site of production for the second hormone is the gonads specifically the ovaries. In this case, hCG is stimulating the production of progesterone from the corpus luteum. Chorion is a fetal/placental tissue where the hCG is derived from. Figure 28.18 (p. 1100) illustrates how hormone levels change in the blood of a pregnant woman. Clinically, hCG can first be detected in serum or urine of approximately 5% of the pregnant women by 7-8 days after conception, and in virtually all women by day 11 (prior to the first missed period). These tests can be very sensitive and detect as little as 5 mIU (IU = International Units; this is the preferred units for hormones, especially when the mass amount is unknown, but a biological effect can be observed and measured). (Not all home pregnancy tests are this sensitive.) In general, hCG levels double about every 2-3 days in early pregnancy. Human chorionic gonadotropin levels rise and peak at about 43-64 days. Most over the counter, home pregnancy tests typically detect approximately 25mIU and are usually used after the woman misses her first menstrual periodabout 10-15 days after ovulation. This is late enough in the pregnancy for sufficient hCG levels to be present. How do commercial pregnancy tests work? Antibodies are plasma proteins of the globulin class (specific blood proteins) that your body produces in response to an antigen (a molecule capable of inducing an immune response). Antibodies will bind and react with antigens. (These are standard immunological terms that are frequently used in a clinical setting.) Pregnancy tests are called ELISA (enzyme-linked immunosorbent assay) "sandwich assays" meaning that two antibodies, collected from the serum of different test animals in the lab, capture hCG between them and in doing so create a colored compound that indicates a positive test. One antibody, the capture antibody, is permanently attached to a membrane like the absorbent paper inside the pregnancy test strip. The second antibody, the tracer antibody, has an enzyme chemically bound onto it and is mobile in the liquid phase of the. Urine, or blood, is added to the assay system; incubated a short time and if

hCG is present it will bind to both the capture and tracer antibodies linking them together into a specific location. In this way, the hCG is sandwiched between the two antibodies. Because 1) the capture antibody is attached to the membrane in a given pattern (sometimes as a + sign), 2) the hormone attaches to the capture antibody, and 3) grabs the enzyme-laden tracer antibody, and the attached enzyme catalyzes the production of a colored, immobile dye, this entire procedure will greatly amplify the presence of a few hCG molecules into many dye molecules in a very limited area on the membrane and typically is very sensitive for confirming pregnancy. Examine a test strip in detail. Note that there is a spacer (pill-shaped structure in the inferior portion of the strip). It is primarily designed to control the humidity of the strip. There is a sheet of absorbent paper that runs the middle portion of the strip. This is where the stationary -- capture antibody is attached. The tracer antibody will dissolve in the urine and bind to hCG if present. If hCG is present, it will bind to the stationary capture antibody and the second color antibody will be concentrated with the mobile hCG and form a line. If no hCG is present, the positive experimental line (pink) will not show up. An additional positive control line is also present on the strips. This region contains immobile hCG that is bound to a separate part of the membrane. Tracer antibodies binding to hCG in this area generate a dye color to indicate that the test strip is working and able to show a positive response. If no dye line shows on the test strip, this indicates that the test did not work, so you cannot conclude anything about hCG levels in the sample. A false negative reading typically means that the test was done too early in a pregnancy. Most false negatives are due to experimental error. In a few cases a false positive signal can occur. Again the most common reason is experimental error although there are some medical conditions that can produce elevated hCG levels that will be detected by the test. For example some liver cancers can produce high levels of hCG. Citations:

1. Advanced Fertility Center of Chicago.

http://www.advancedfertility.com/earlypre. htm. 2. Quidel Corp. Quick Vue -- one step hCG combo test. San Diego, CA 92121. 800874-1517. 3. Rhodes, S. J. and L. R. Banner. 2001. Are your cells pregnant? Relating biology laboratory exercises to everyday life. The American Biology Teacher 63 (7): 514-517.

4. University of New Mexico. Health Sciences Center and School of Medicine. Womens Health Research. http://www.hcglab.com/hcgtest.htm. 5. Weiss, R. L. 2001. The rabbit died! The history of pregnancy test from rabbits to hCG. http://pregnancy.about.com/library/weekly/ aa090901a.htm

Test Procedure for Qualitative Detection of hCG: 5 volunteers will be asked to donate urine. The lab instructor will randomly select 3 samples from all samples available (the positive control urine was donated by a pregnant woman, the negative control was donated by Dr. Wilson, who is not pregnant.) As a demonstration three drops of sample urine will be added to each round sample well on the pregnancy test strip. What result will you expect for the Positive and Negative control? Samples are allowed to incubate 3 minutes to dissolve the liquid phase antibody and allow the assay to work. Be sure not to move or shake the kits while incubation. Each sample will be assessed as to being a positive indication or a negative response. A positive response will be a bar (either red, pink, pale peach) across the T designated area. The blue bar across the C area will always be present if the test works correctly.

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