James Kashima

Neurobiology 302
Lab 7: JAR

Abstract
The purpose of this experiment as to record and identify various parameters of the
Jamming Avoidance Response (JAR) seen in weakly electric fish. This response changes
a fishs frequency of electric organ discharges (EODs) when another fish with a similar
EOD frequency is nearby, allowing it to continue to use its own EOD for such important
functions as electrolocation. The fish we used, A. albifrons, accurately responded to a
wide range of stimulus frequencies including those approximately 35 Hz away from its
own natural frequency, and that the time course of the JAR response was such that it was
extremely sensitive to the onset of a stimulus and took a comparatively longer time to
return to close to its natural frequency. We postulate that this slow time course of the
recovery phase of the JAR is a type of energy saving mechanism utilized by the fish in
areas that may be highly populated by other individuals of the same species.

Introduction
A. albifrons is a species of weakly electric fish found in the freshwaters in Central
and South America. An electric organ found in the caudal end of the animal sends out
electric organ discharges, or EODs to communicate, navigate, and sense potential
predators and mates. In order for this to happen, the fish also has electroreceptors
(ampullary and tuberous) to detect electric both its own EODs and electric signals
coming from other electric fishes and organisms. These receptors are able to pick up
minute changes in phase and amplitude of a returning signal, allowing the fish to know its
surroundings. Because these fish rely so heavily on being able to pick up their own EODs
for electrolocation, it is essential that their own signal does not get mixed up with the
EOD coming from another individual, which could hamper the fishs ability to navigate.
Each fish has a unique frequency of EODs that it fires at, but if it is nearby another
individual with a similar EOD frequency, the two fish will elicit a Jamming Avoidance
Response (JAR) to increase the difference between their two EOD frequencies so that
their signals do not interfere with each other; the fish with the higher EOD frequency will
increase its frequency, and the fish with the lower EOD frequency will shift its frequency
down. This is all done unconsciously by the fish via a complex circuit involving the
electroreceptors, cells in the electrosensory lateral line lobe, torus semicircularis, nucleus
electrosensorius, prepacemaker nucleus, and pacemaker nucleus. Despite the differences
between the senses we use to navigate and those employed by A. albifrons, it is important
to be able to understand how different organisms perceive and interact with the world,
and how organisms may process various inputs in their central nervous systems.

Methods
The work in this lab report followed the protocol outlined in the lab manual
(Bosma, 2013). After setting up the fishtank with the fish inside with the proper recording
electrodes, we first recorded the fishs natural EOD frequency using the Labchart
computer software to take the EOD voltages and make a Fourier power transform to give
us the frequency. We then placed our stimulating electrodes into the water and stimulated
the fish with an EOD frequency close to its own and with a lesser amplitude. Following
this, we maintained the stimulating EOD amplitude but varied its frequency in a range
both above and below the fishs EOD frequency. Throughout the experiment, the water
temperature was maintained at 29.5C +/- .5C.

Results
The fishs normal EOD traces are seen in Figure 1. The amplitude of each EOD
was around .57 V, and each EOD lasted for around .8 ms. Figure 2 shows a Fast Fouier
Transform of the data from Figure 1. Frequency is shown on the x axis and power is
shown on the y axis. The largest peak, which corresponds to a frequency of 1198.3 Hz, is
the value we use as the spontaneous EOD frequency of the fish, as this was the most
common frequency throughout the spontaneous activity. Figure 3 shows the extracellular
recordings in the tank during the application of an artificial stimulus. Because the
recordings electrodes are placed inside the water, they pick up all electrical activity inside
the fish tank. Figure 3s shape is caused by the summed amplitudes of the two EODs (the
fishs and the artificial), and its amplitude changes in a manner known as the beat
frequency, which is the difference between the frequencies of the fish and of the artificial
stimulus. The beat frequency in Figure 3 is about 16 Hz, and the combined EOD
frequency is 1216.5 Hz.
Figure 4 shows how the JAR changes over time, plotting the EOD frequency over
time. The stimulus frequency was 8 Hz below that of the fishs. The stimulus started
immediately after time -178.5 s and ended at time 0 s. Prior to the stimulus, the fishs
EOD frequency was at 1192 Hz, and after the onset of the stimulus, the fish increased its
EOD frequency to 1202 Hz. However, the maximum EOD frequency occurred after the
stimulus was shut off, with a peak frequency of 1217 Hz. After the stimulus onset, the
fish quickly increased its EOD frequency, but it took more than 100 seconds for it to
decrease its frequency to levels close to its stimulus frequency.
Figure 5 shows a graph of R, (change in the fishs own frequency before
stimulus minus its frequency during the stimulus) plotted as function of F (The
frequency of the artificial stimulus minus the fishs frequency before stimulus). The
points show that the fish responded with normal JAR behavior, changing the frequency of
its own EODs so that the difference between its EOD frequency and that of the stimulus
increased. Generally, when the F was smaller, the response in R was larger, meaning
the change in the fishs EOD frequency was of a smaller magnitude.

Discussion
The waveforms exhibited in Figures 1 and 3 show that the fish changed its EOD
frequency in response to an artificial fishs EOD, as was expected. The neural basis of
JAR, elucidated by Heiligenberg, indicates that the fish uses differential geometry of the
incoming signal to determine in which direction to shift its EOD frequency. It does this
by separately comparing the phase and amplitude modulation occurring in two different
parts of its body and sending this information to sign selective cells in the torus
semicircularis, which then send a signal to 2 sign selective cells in the nucleus
electrosensorius that can excite or inhibit the prepacemaker nucleus which ultimately
alters the frequency of the EODs by signaling to the pacemaker nucleus (Heiligenberg
1989). Differential geometry was achieved in the tank as the stimulating electrodes were
placed on one side of the fish. If the electrodes were placed such that they bisected the
fish rotral to caudal, then no EOD would be expected as there would be a lack of
differential geometry.
The time course of the JAR in Figure 4 showed an interesting trend. The EOD
frequency of the fish jumped up immediately to 70% of its maximum JAR frequency,
taking 1.5 seconds to reach 80%, and 7 seconds to reach 90%. However, once the
stimulus was shut off, an interesting phenomenon was observed, as the EOD frequency
immediately jumped up very high to a frequency much higher than that obtained with the
stimulus on. It took 11 seconds for the frequency to decrease to the stimulus level, and
then a total of 95 seconds to return to close to the pre-stimulus level. Thus, the time
course of the recovery phase was markedly slower than that of the stimulus phase. The
extremely large increase in the EOD frequency after the stimulus was shut off could have
been an artifact of disconnecting only one alligator clip at a time, or it could be possible
that the fish reacted to the sudden drop in the other fishs EOD to mean that it had to
increase its own EOD even more. In relation to the longer time course of the recovery
phase, this could be hysteresis on the part of the fish. If an electric fish comes close
enough to another fish with a similar enough EOD frequency to elicit a JAR, then the fish
will most likely have a way of remembering that a fish in its nearby vicinity has a
similar frequency to its own. Thus, after it elicits a JAR to that fish, its EOD probably
will not decrease all the way back to its normal levels for some time. If both fish do this,
then the next time they come close to one another, no JAR will occur, as both fish will
have changed their EOD frequencies to be a little further from what they were previously
so that no effort is put into a JAR. The fast initial rate of decay of the recovery phase
could be so that the fish does not have to expend excess energy firing EODs at a much
greater-than-normal rate. Throughout the stimulus, it is very important for the fish to not
allow jamming of its electroreceptors from the other fishs EODs, so it works quickly and
effectively to greatly raise the frequency of its EODs. However, once the signal from the
other fish disappears, the immediate threat of jamming disappears as well, so the fishs
EOD frequency can move into a less cautious state of firing and fire closer to its natural
frequency. This hypothesis could potentially be related to the results in Figure 5. In
general, the closer the artificial EOD frequency is to the fishs natural EOD frequency,
the greater the frequency modulation exhibited by the fish. In a sense, the fish works
extra hard to make sure that its electrolocation abilities are not compromised by the
EODs of the other fish. As the F gets larger, the fish has to work less hard in the sense
that the frequency modulation it must perform decreases, seen by the decreasing R
values. This would also seem to indicate a threshold F value after which the fish does
not JAR in the presence of a stimulus. What this threshold value is and how it is
determined neutrally would be interesting work for future experiments, and would
increase our understanding of how these fascinating electric fish work.
















Figures:

Figure 1: EOD frequency of fish: The EOD frequency of the fish as it swam in the water is shown. The baseline frequency is
maintained at around 4Hz, but this is at times interrupted by peaks in frequency that then decrease back to baseline levels.


Figure 2: EOD frequency with aluminum plate oriented parallel to fish: This figure shows the EOD frequency in response to the
aluminum plate oriented parallel to the fish. The only spike in frequency came when the plate was moved away from the fish.


Figure 3: EOD frequency with aluminum plate oriented perpendicular to fish: This figure shows the EOD frequency in response
to the aluminum plate oriented perpendicular to the fish. The frequency increased as the plate approached the fish and as it
moved away from the fish, but not while it was next to the fish.

Figure 4: Cross-correlogram of another fish compared to our fish: This cross-correlogram shows the EOD response of another
fish in relation to our fish. The other fish appeared to fire many EODs about 27 ms after our fish sent out an EOD, indicating a
possible echo response of a dominant fish.


Figure 5: Peristimulus time histogram of our fishs EOD response to a computer stimulation: This PSTH shows how our fish
responded to an electrical signal sent by a computer of 5 V. Our fish sent responded between 10-20 ms after the computer sent out
its artificial EOD, possibly indicating an echo response, meaning our fish was attempting to exert dominance over the computer.





















References

Heiligenberg, W (1989). Coding and Processing of Electrosensory Information in
Gymnotiform Fish. Journal of Experimental Biology 146: 255-275

Bosma, Kennedy, Olavarria (2013). Introduction to Systems and Behavioral
Neurobiology. Course Manual: 63-70