Renal Physiology & Osmoregulation Report

Johanna Coutinho
&
Ezgi Ertemin-Pearson
BSC2086C Lab, Section 50211
July 11, 2014

Results
The three graphs located in the following data section show the trends discussed
in this section for three separate test groups: the hypotonic group, which ingested 650ml
of water; the isotonic group, which ingested 650ml of Gatorade; and the hypertonic
group, in which each subject ingested 80ml of water, plus 1g salt/20kg of body weight.
The results that were obtained showed discernible differences among the three test
groups, in response to the tonicity of the solutions ingested. These differences were
expressed in urine volume, the specific gravity of the urine samples, and their respective
salt concentrationsspecifically, the sodium chloride content of the urine.
Figure 1 shows the trend of urine output over time, expressed in milliliters per
minute. Urine volume increased for all three test groups within the first 30 minutes, but
the graph indicates dramatic changes in output thereafter. The isotonic group had the
most rapid increase in urine volume, rising from 0.9 to 2.8ml/min. However, the
hypotonic group caught up at the 30-60 minute interval, where both groups peaked
almost identically (3.3ml/minute for the hypotonic group and 3.4ml/minute for the
hypotonic group). At the same time, the hypertonic group began demonstrating the polar
opposite effect, as those subjects took a dramatic turn at 60 minutes and decreased from
an average of 2.4 to 0.6ml/min in urine output. The hypertonic group steadily decreased
by 0.01ml/min every 30 minutes thereafter, maintaining very low urine volumes. Both
the isotonic and hypotonic groups also decreased in urine volumes, but the isotonic group
remained more constant. Between 90-120 minutes, the hypotonic group decreased their
average volume by half, from 2.8 to 1.4ml/minalmost as dramatic a change as they
experienced during the 30-60 minute interval. By the end of the experiment (120

minutes), the hypotonic group was still producing more urine per minute than before they
had ingested the water, averaging 1.4ml/min instead of 0.5ml/min. The isotonic group
was also producing more than double the urine volume they had been, prior to drinking
Gatorade (0.9 to 2.2ml/min). The hypertonic group, in contrast, was only producing a
little over 30% of their original urine volume (0.4ml/min, compared to 1.1ml/min).
The specific gravity measurements of the test subjects indicated solute
concentrations of the urine samples provided during each 30-minute interval, as shown in
Figure 2. All three groups began with very similar averages before ingesting their
respective solutions, but over time, each groups specific gravity changed in unique ways.
The isotonic groups specific gravity barely shifted within the first 30 minutes, while both
the hypotonic and the hypertonic groups experienced a slight decrease. From that point,
on, however, all the test subjects followed different trends. The hypotonic group had a
dramatic decrease in specific gravity, from 1.017 to 1.005. The isotonic group decreased
slightly, from 1.014 to 1.011, and the hypertonic group had a significant increase in
specific gravity, from 1.012 to 1.022. Between 60-90 minutes, there were no dramatic
shifts, but the hypertonic group continued to steadily increase, while the hypotonic group
slightly decreased in specific gravity. The isotonic group, meanwhile, remained the same.
Finally, during the final 90-120 minute interval, the hypotonic group began to rise again,
from 1.003 to 1.009. The hypertonic group still continued to climb, reaching a final,
dense value of 1.025 at 120 minutes. The isotonic group increased slightly, as well, with
a final value of 1.013.
The trends in Figure 3 express sodium chloride concentration in the urine. Both
the hypotonic and the isotonic groups had an immediate decrease in sodium chloride

concentration 30 minutes after ingesting their respective solutions, while the hypertonic
group experienced no change, overall. At 60 minutes, however, the hypertonic group
experienced an impressive increase, from 5.6 to 8.7mg salt per milliliter of urine. The
isotonic group continued to decrease steadily, from 5.4 to 4.5mg/ml, while the graph
shows a steeper decline for the hypotonic group, from 5.3 to 2.6mg/ml. While the salt
concentration for both the hypotonic and hypertonic groups seemed to slowly increase
over time for the remainder of the experiment, the isotonic group experienced more
fluctuations in their results. After having decreased in concentration for an hour, the urine
of the isotonic group suddenly surged back up to 5.5mg/ml after 90 minutes, but by the
end of the experiment, the concentration was once again declining, with a final value of
5.1mg/ml.

Data

Discussion
The kidneys are one of our most important organs to maintain body homeostasis
through the regulation of blood volume and concentration (Marieb & Hoehn, 853). To
keep the normal osmotic concentration300 mOsmthe kidneys osmoregulate our
body fluids from becoming too diluted or too concentrated.
As seen in the Figure 1, urine volume of different groups changed over time
according to the osmolality of the substances that they ingested. The hypertonic group
that ingested coated salt tablets had an increase in the urine volume within the first 30
min due to the small amount of water they ingested with their salt tablet and their preexisting blood pressure. However, a dramatic decrease in urine output occurred between
the 30-60 minute interval. Ingesting a high amount of salt increased the sodium content
of their filtrate; as a result, their extracellular fluid became hyperosmotic. Antidiuretic
hormone (ADH) is released from their posterior pituitary in response to hyperosmotic
extracellular fluid (Marieb & Hoehn, 850). ADH inhibited their urine output by inserting
aquaporins into the apical membranes of their collecting ducts. Collecting ducts that are
permeable to water, only in the presence of ADH, aid in reabsorption of water to
compensate dehydration caused by excess salt intake.
Another factor that contributed to the decrease in urine output in the hypertonic
group was the renin-angiotensin-aldosterone mechanism. Macula densa cells located in
the ascending limb of the nephron loop that monitor NaCl content entering the distal
convoluted tubule (DCT) signaled granular cells to secrete renin. Renin enzymatically
started the conversion process to produce angiotensin-II, and angiotensin-II then
stimulated the adrenal cortex to release aldosterone hormone. Aldosterone hormone fine-

tuned the reabsorption of the remaining sodium (Marieb & Hoehn,851). As sodium
reabsorbed at the DCT and collecting ducts, water followed the sodium and left the
collecting duct via facultative water reabsorption provided by ADH. As a result of the
renin-angiotensin-aldosterone mechanism, urine volume decreased.
The hypertonic group continued to display a volume decrease steadily between the
60-90 minute interval, as well as between the 90-120 minute interval. However, the
decrease was not as dramatic as it was between 30 and 60 minutes. Because urine volume
was already very low between the 30 and 60 minutes due to dehydration, the results that
followed remained consistent, as the participants did not rehydrate themselves to provoke
a change in urine volume.
On the other hand, the urine volume trend line of the hypotonic group who ingested
water was very different than the hypertonic group. Although they both had an increase
in urine volume after 30 minutes, the difference in urine volume after 60 minutes was
dramatic. The ADH and aldosterone decreased in the hypotonic group, following the
decrease in their osmolality of extracellular fluids. As a result, water permeability of the
collecting ducts decreased as well and water was retained in the ducts. Their kidney
produced a high volume of urine to compensate for over-hydration. After 60 minutes,
their urine volume started to decrease because as the participants micturated, their
extracellular fluid decreased. They also did not ingest water until before the 120th
minutes data was recorded.
The isotonic groups urine volume trend line displayed similar patterns to the
hypotonic group. Their urine volume steadily increased and reached its peak at the 60-

minute interval and continued to drop, thereafter. Because the fluid that they ingested was
Gatorade, which contained carbohydrates, sodium, and potassium, these solutes helped
maintain an equal osmotic pressure with their bodies. For this reason, ADH and
aldosterone levels also decreased in the isotonic group. Although the isotonic group had
similar trend lines with the hypotonic group, they had higher urine output. This was also
related to the well-hydrated participants of the isotonic group.
Specific gravity is the ratio of the mass of a substance to the mass of an equal
volume of distilled water. (Marieb & Hoehn, 858). The specific gravity measurement is
taken by an urinometer that determines the solute concentration. The urinometer calibre
marks 1.000 for distilled water, since it is free of any solutes. As urine becomes
concentrated, its specific gravity increases; as it becomes more dilute, specific gravity
decreases. We can, therefore, conclude that there is an inversely proportional relationship
between urine volume and specific gravity.
The hypertonic groups specific gravity decreased after 30 minutes, as their urine
volume increased. Their specific gravity continuously increased thereafter, as their urine
volume decreased and became more concentrated. The juxtamedullary nephrons were
activated, with nephron loops that drop into the medulla of the kidney, to adjust their
solute concentration according to the osmotic gradient established by countercurrent
mechanisms that determines urine concentration and volume. Also, their increased ADH
and aldosterone contributed to their increased specific gravity.
The hypotonic groups specific gravity decreased after 30 minutes because their
urine became more dilute; ADH and aldosterone were decreased and, as a result, their

urine concentration decreased, as well. After 90 minutes, their specific gravity displayed
an increase because their urine volume had dropped.
The isotonic groups specific gravity displayed similar trend line fluctuations as the
hypotonic group. This was because their urine volume was similar. However, their
specific gravity was always higher than the hypotonic group due to the solute content
ingested in the Gatorade. Their specific gravity also increased between the 90-120 minute
interval as a result of decreased urine volume.
All three groups specific gravity results were in a healthy range, between 1.001
and 1.030 (Marieb & Hoehn, 858). There were no participants who passed 1.030, which
could indicate a pathology. However, there were many low specific gravity results. One
of the reasons for this was over-hydration. Also, in some cases, participant were not able
to determine the specific gravity due to low urine volume that interfered with the
urinometer measurement.

In chemistry, titration methods are based on finding the unknown concentration of a

substance by adding a standard solution with a known concentration. Silver nitrate was
our known substance and it had a 2.9 % molarity. Each drop of silver nitrate
corresponded to 1 mg/ml of sodium chloride that was present in the urine sample.
Counting how many drops of silver nitrate added to the urine sample to change its color
to dark red- brown color indicated its sodium concentration.
The hypertonic group had the highest sodium chloride concentration as a result of
ingesting a salt tablet. Their salt concentration had a dramatic peak after 60 minutes and

continued to increase slowly. Sodium chloride concentration trend lines correlated with
their specific gravity trend lines because sodium and chloride ions increases the amount
of solutes present in the urine. Sixty-five perecent of their sodium reabsorbed in the
proximal convoluted tubule (PCT) via the sodium-potassium pump located on their
basolateral surface. While sodium is reabsorbed by primary active transport, chloride ions
followed its electrochemical gradient (passive paracellular diffusion). Countercurrent
multiplier interactions between the ascending and descending nephron loops of the
juxtamedullary nephron determined the osmolality of the filtrate and the sodium
concentration entering the DCT. ( Marieb & Hoehn, 853). Hormonal mechanisms,
previously discussed to explain urine volume, determined the final sodium concentration.
Decreased urine volume was the cause for the sodium concentration that is present in the
urine volume.
The hypotonic group had a dramatic decrease in sodium chloride concentration at
the 60-minute interval, as their urine volume reached its peak. Since their urine was
extremely diluted due to over-hydration, their sodium concentration was at its lowest.
However, it increased continuously after 60 minutes, while their urine volume decreased.
The isotonic groups sodium chloride concentration trend line was similar to that of
the hypotonic group. However, the isotonic group had a higher sodium concentration in
their urine because Gatorade had sodium, as well. The isotonic group also displayed its
lowest sodium concentration after 60 minutes and continued to rise, as their urine volume
decreased.
Changes in salt concentration and urine volume show the kidneys homeostatic

balance mechanism at play. Kidneys adjust urine concentration and volume to keep
osmotic concentration at 300 mOsm. To be able to maintain a steady osmotic pressure,
kidneys utilize countercurrent mechanisms. Also, kidneys regulate the glomerular
filtration rate and arterial blood pressure via intrinsic and extrinsic control mechanisms to
maintain extracellular homeostasis and steady blood pressure.

Reference
Marieb ,E. N, Hoehn K. The Urinary System. In 5th ed. Anatomy and Physiology.
Pearson/Benjamin Cummings. 2010: 834-864.