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Basic Medical Sciences, Kulliyyah of Medicine, International Islamic University of Malaysia

THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS
TOPICS Page

Pharmacology : Autonomic nervous system agonists and antagonists Pharmacology : Routes of administration Elective Practicals

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NAME MATRIC NO

The femoral vein is cannulated for administration of the drugs to be studied. repeat procedures la and lb. 2a. Phenoxybenzamine 3mg/kg is injected. 4. and repeat lb. IIUM © RHM | 18 .) and the trachea is cannulated. NAME MATRIC NO Objectives: To study the effects of Sympathetic and Parasympathetic agonists and antagonists on the anaesthetized cat. the basal BP. Noradrenalin 3 µg/kg. heart rate.p.THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS AUTONOMIC NERVOUS SYSTEM : AGONISTS AND ANTAGONISTS DATE GROUP MEMBERS NO 1. and contraction of the nictitating membrane is recorded. The heart rate can also be recorded simultaneously. Inject Propranolol 2mg/kg and wait for 10 minutes. lb. In order to record the blood pressure changes. 3. Next. For recording the movements of the ileum. can also be recorded via a transducer. In the 2nd cat. Sympathetic Cat la. The nictitating membrane of the eye contains aipha-adrenergic receptors and its contraction on stimulation. the carotid artery is cannulated and connected to a pressure transducer. Wait for 10 minute. Department of Basic Medical Sciences. Experiment 1. The vagus nerve is identified and is prepared for electrical stimulation. stimulate the nerve to the nictitating membrane and repeat lb. 2b. When the conditions have stabilised in the l cat preparation. Procedure: The cat is anaesthetized with Pentobarbitone (40mg/kg i. 1 d. a balloon filled with water placed in the ileum of the cat and is then connected to a transducer. Adrenaline 3 µg/kg and Isoprenaline 3 µg/kg are injected one after another and their effects on BP and HR are recorded. 2. Ic. Kulliyyah of Medicine.

Inject Carbachol 2µg/kg and record the results. IIUM © RHM | 19 . the BP. Inject Nicotine 200µg/kg and record the results. All students are required to enter their results with a short explanation of the results in the attached schedule list and submit them to the lecturer immediately for assessment. 3c.THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS Experiment 2. 3b. 3f. Department of Basic Medical Sciences. HR and gut motility. Inject Acetylcholine 2µg/kg and record the results. 3c. 3f. 3e. and 3d and record the results. Kulliyyah of Medicine. 3g. Inject Acetylcholine 300µg/kg and record the results. Repeat procedures 3b. HR and the contractions of the ileum are recorded. Stimulate the vagus nerve and record the BP. 3d. When conditions in the 3rd cat have stabilised. Parasympathetic Cat 3a. Now inject Atropine 2mg/kg and wait for 10 minutes.

THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS PARASYMPATHETIC CAT Blood Pressure Systolic/Diastolic Vagus Stimulation Control Test Acethycholine 2µg/kg Control Test Carbachol 2µg/kg Control Test Atropine3 mg/kg Control Test Acethycholine 2µg/kg Control Test Carbachol 2µg/kg Control Test Acethycholine 300µg/kg Control Test Nicotine 200µg/kg Control Test Mean Arterial Heart Rate ( BPM) Gut Motility Department of Basic Medical Sciences. IIUM © RHM | 20 . Kulliyyah of Medicine.

Membrane Contraction Department of Basic Medical Sciences. Kulliyyah of Medicine.THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS Sympathetic Cat 1 Blood Pressure Systolic/Diastolic Noradrenaline 3µg/kg Control Test Adrenaline 3µg/kg Control Test Isoprenaline 3µg/kg Control Test Phenoxybenzamine 3 mg/kg Control Test Noradrenaline 3µg/kg Control Test Adrenaline 3µg/kg Control Test Isoprenaline 3µg/kg Control Test Mean Arterial Heart Rate ( BPM) N. IIUM © RHM | 21 .

Kulliyyah of Medicine. Membrane Contraction Control Test Noradrenaline 3µg/kg Control Test Adrenaline 3µg/kg Control Test Isoprenaline 3µg/kg Control Test Department of Basic Medical Sciences. IIUM © RHM | 22 .THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS Sympathetic Cat 2 Blood Pressure Systolic/Diastolic Noradrenaline 3µg/kg Control Test Adrenaline 3µg/kg Control Test Isoprenaline 3µg/kg Control Test Propranolol 2 mg/kg Mean Arterial Heart Rate ( BPM) N.

Each of these routes has its particular advantages. Note the time of administration of the drug and observe the animal for its effect. intravenous. the character. to a large extent. Place the tail from the right hand to the left small finger so that the mouse is held tightly in the left hand. intramuscular.5m1/10 gm of animal may be administered. Introduction The common routes of administration of drugs are oral. IIUM © RHM | 23 . intrathecal. dose and degree of action. The mouse is now ready for injection. grasp the nape of the animal’s neck between the thumb and the index finger. Righting attempted without success. and allow it to grip the wire mesh of the cage with its forelegs. The mouse is held firmly and the feeding needle is held well towards the back of the mouth and near the upper palate. 2. and topical application. to Department of Basic Medical Sciences. With the left hand. As much as 0. NAME MATRIC NO General Instructions Students will be assigned to work in groups. Handling of Mice Lift the mouse by grasping the tail at the caudal end with the right hand. speed. rectal. No locomotion response to stimulation. Absence of spontaneous activity with uncoordinated locomotion upon stimulation. 3. The needle is than passed-gently-not-forced down the esophagus. Each group is given 10 mice. The mode of entry of a drug into the body determines. ORAL ADMIINISRATION Administer by a stomach tube or by an oral feeding needle 35 mg/kg (0. Kulliyyah of Medicine. subcutaneous. 2. 4. Minimal spontaneous activity with normal responsiveness to stimulation. Note the several grades of response as follows: 1. As it passes into the stomach (about 2 inches down) the animal will exhibit a definite and characteristic gagging.1 ml/l0 gm body weight) of pentobarbital sodium (Nembutal) to a mouse. 4. nasal. but righting reflex retained. A.THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS ROUTES OF ADMINISTRATION DATE GROUP MEMBERS NO 1. 3. Some substances may be effective when administered by one route but relatively or completely inert when given by another.

IIUM © RHM | 24 . Inject 35 mg/kg (0. Employ a sharp. D. record the nearest minute to the time of onset and the duration of loss of the righting reflex. One must always pull back on the plunger to make certain the needle is not in a vein Make your observation and record the results in the table provided. The drug is injected into the peritoneal cavity where absorption is rapid.1 ml/l0 gm of body weight) of pentobarbital sodium into the mouse. intraintestinal or intravisceral injection may be avoided. pool the data obtained from each student in your lab.1 ml/l0 gm of body weight) of pentobarbital sodium under the skin of a male mouse followed by gentle massage of the site of injection. At the end of the session. With a little practice the tail veins can be recognized readily. SUBCUTANEOUS INJECTION The drug is to be introduced directly underneath the skin. Department of Basic Medical Sciences. If the needle is not in the vein. A 3/4 -l inch 26-gauge needle is employed. compare effects with those noted in previous sections. using a 26-gauge needle. Immobility and righting not attempted In the table. and record the data in the table provided. resistance will be felt while injecting and blanching will be observed in the surrounding connective tissue.THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS 5. INTRAMUSCULAR INJECTION Inject 35 mg/kg (0. B. C. Observe as under A. INTRAVENOUS INJECTION The mouse is to be held firmly by wrapping it in a cloth or by placing it in a suitable animal holder. It is helpful to dilate the veins by warming before attempting the injection. Insert the needle to its full length to avoid loss of liquid upon withdrawal. Inject 35 mg/kg (0. Kulliyyah of Medicine. The common site employed is the dorsal neck area and that of the abdomen.1 ml/ 10 gm of body weight) of pentobarbital sodium into the mouse and record your observations. Obtain the mean and standard deviations of the values that your lab group has obtained.1 ml/l0 gm of body weight) of pentobarbital sodium into the gluteus maximus muscle of a mouse. INTRAPERITONEAL INJECTION This is a common method to introduce drugs into animals. By holding the animal’s head downward. inject into the lower half of the abdomen. E. 26-gauge needle and slowly inject 35 mg/kg (0. and record the data in the table.

THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS EFFECT OF DIFFERENT ROUTES OF ADMINISTRATION Route of Administration Group Onset Oral Subcutaneous Intravenous Intraperitoneal Intramuscular Data Duration Class Onset Average Duration For group. IIUM © RHM | 25 . Department of Basic Medical Sciences. calculate: 1. Mean duration of action ± standard deviation Comment and compare the time of onset and duration of action of each route of administered. Kulliyyah of Medicine. 2. Mean time of onset ± standard deviation.

Kulliyyah of Medicine. Department of Basic Medical Sciences. The Ambulatory 24-hour blood pressure monitor. and blood pressure will be monitored as you exercise (using a treadmill) at increasing levels of intensity. Following the measurement. IIUM © RHM | 26 . All you have to do is to take a few simple steps: 1. The devices are : 1. 4. 2. ECG pattern. 24 hour Ambulatory blood pressure monitor. your BP is monitored throughout the 24 hour duration (even during solat and in sleep). 2. Your blood pressure is measured. and the blood pressure cuff is left in place to allow regular blood pressure measurements during the test. Stress test. As the name implies. you will receive a copy of your results. The stress test ECG leads are attached to your chest. To answer a questionnaire on the day of measurement. Your heart rate. you are invited to have cardiovascular parameters measured using the devices which are commonly used in patients’ management and in research.THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS ELECTIVE PRACTICALS In this section. This will require you to wear a blood pressure monitoring for 24 hours (Yes! you are allowed to take the devices back to Mahallah). There will be certain rules to follow while having the measurement taken. To call our Medical Lab Technologist Br Shahriman (ext 2382) to make an appointment for the measurement. To see any Pharmacology or Physiology lecturers to discuss on the results. 3.

1. What is the clinical indication for the stress test? ____________________________________________________ 3. Who will benefit from having their blood pressure measured by this way? ______________________________________________________________ ______________________________________________________________ ______________________________________________________________ _____ 3. What is the information you can get from the stress test? ____________________________________________________ 2.THE PRACTICAL BOOK FOR PHARMACOLOGY & PHYSIOLOGY LABORATORY EXPERIMENTS SELF LEARNING Stress test 1. IIUM © RHM | 27 . what can ABPM tell us? ______________________________________________________________ ______________________________________________________________ ______________________________________________________________ _____ Department of Basic Medical Sciences. especially for patient? ______________________________________________________________ _ The Ambulatory 24-hour blood pressure monitor (ABPM). Can you compare the information that we can get from using ABPM to that of using sphygmomanometer? ______________________________________________________________ ______________________________________________________________ ______________________________________________________________ _____ 2. In a drug clinical trial. What is the necessary precaution in performing this test. Kulliyyah of Medicine.