EXPERIMENTAL PROTOCOL FOR SYSTEMIC ANF
                INFUSION STUDIES IN THE NORMAL RAT

1) DAY BEFORE THE EXPERIMENT ONE SHOULD PREWEIGH TESTTUBES FOR 
        URINE SAMPLES. WEIGH RATS.
2) PREPARE INULIN AND PAH INFUSION SOLUTIONS IN ADVANCE (DAY OF EXPERIMENT).
        TOTAL VOLUME = 30ML OF SOLUTION (NORMAL SALINE) ONE ADDS TO THESE
        30ML OF THE FOLLOWING AMOUNTS OF INULIN (IN MG) = 180 * GM OF RAT/100
                                         PAH    (IN MG) =  30 * GM/100

        AFTER DESOLVING INULIN AND PAH IN SALINE SOLUTION AND COOLING TO ROOM 
        TEMPERATURE YOU CAN START INFUSION AT 33.34 ul / min. WAIT 1 TO 1.5
        HRS FOR EQUILIBRIUM OF PAH AND INULIN BEFORE BEGINNING CLEARANCE
        STUDY.
3) ANF BOLUS I.V. = 2ug/kg
        ANF STOCK SOLUTION = 10ML OF NORMAL SALINE; ADD 60.8 ul OF ANF STOCK
         TO GET 1.216 ug/ml STOCK. EXAMPLE FOR  A 150 GM RAT YOU GIVE 246.7 ul
         OF STOCK.

4) ANESTHETIZE RATS WITH ETHER. THE RAT MUST BE AWAKE BEFORE BASAL CLEARANCE
        IS TAKEN. TO ANESTHETIZE RAT WE HAVE OBTAINED GOOD RESULTS USING A
        PLASTIC SYRINGE HOLDER WITH A GAUZE SPUNGE TO RETAIN THE EHTER. PLACE 
        OVER HEAD OF RAT TAKING SPECIAL CARE NOT TO OVER DO IT. IF RE- 
        SUCITATION IS NECESSARY, HAVE A SMALL PLASTIC STRIP OF POLYETHYLENE  
        TUBING AVAILABLE; PLACE OVER NOSE AND MOUTH OF RAT. 
5) CANULATE ONE CAROTID ARTERY
   ONE JUGULAR VEIN, AND START PAH AND INULIN INFUSION.
   ONE FEMORAL ARTERY  
   BLADDER.
6) TAKE BASAL (30'INTO THE STABALIZATION PERIOD) URINE AND BLOOD SAMPLE
7) BLOOD PRESSURE MUST BE MEASURED BEFORE AND DURING I.V. BOLUS OF ANF.
8) A CONSTANT INFUSION OF ANF ( 0.3 ug/kg/min) IS STARTED, USING THE STOCK
        SOLUTION .
        EXAMPLE. FOR  A 150 gm RAT YOU NEED  TO INFUSE 0.0345 ug/min OR
                0.037 ml/min  I.V. OF ANF STOCK SOLUTION. 
9) A CLEARANCE PERIOD IS BEGUN WITH B0LUS ADMINISTRATION AND CONTINUED FOR 
        30 MIN.
10) AT THE MIDPOINT OF EACH TIME INTERVAL, TAKE A BLOOD SAMPLE FOR PLASMA 
        RENIN ACTIVITY, HEMATOCRIT AND Na+-K+ DETERMINATION.
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