Ongoing Research:
Primary efforts include characterizing and quantifying male
and female reproductive behaviors of small fish models. Intial work has involved observations of
mummichog (Fundulus heteroclitus) and fathead
minnow (Pimephales promelas). These species are sexually
dimorphic and have several critical reproductive behaviors within
their reproductive behavioral repertoire that can be quantified.
We are interested in examining the range reproductive of strategies
used and determine if differences within this range of reproductive
behaviors, and associated physiological endpoints, effect reproductive
success in terms of maximizing clutch size, fertilization rate,
and survival of offspring. These behaviors include, nest cleaning,
courtship, spawning, egg maintenance and defense (see video clip
below). Once the normal range of reproductive behaviors have
been quantified, fluctuations in environmental parameters that
can alter reproductive behavior and success can then be examined.
The design of our behavior laboratory includes 12 arenas with
CCD cameras (below, left) that transmit data to a multiplexer
and VCRs (below right). Computer-controlled recording allows
for automatic and remote video data collection.
Schematic diagram (above, left) illustrating the behavioral quantification
suite at the University of Maryland Aquatic Pathobiology Center.
A water preparation room with carboys and computer-controlled
pumps provide flow-through or static renewal media support to
the videography room. The videography room contains aquaria that
serve as reproduction arenas, water distribution and drainage
lines (not shown), dedicated CCD cameras set to record from below,
translucent tank dividers (not shown) and shadowless lighting
(not shown). The observation room (above right) contains the
control computer, a video multiplexer, and display monitor. This
equipment permits simultaneous viewing of fish in real time during
an experiment. Video signals are recorded on dedicated VCR decks
that are controlled by X-10 computer software and hardware. Digitized
video data can then have targets (fish) tracked over time (see
movie below), and x,y coordinate data generated for each animal.
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Tracking of fish groups.
(Click on the ""
button at the bottom left of the movie to start movie clip. Movie
may take several minutes to load. If QT is not installed on your
computer, go to http://www.apple.com/quicktime/download.html)
The QuickTime movie on the left shows 5 minnows being followed
by our tracking software as they acclimate over time in a circular
arena. The magenta crosshairs indicate the mid-point of the body.
Based on an algorithm that uses least squares analysis, when
the paths of two fish cross, the tracking software can continue
to follow individual fish with relativey excellent accuracy.
After determining the x,y coordinate data over time for each
of multiple arenas, various questions can be asked of the data.
These questions include trajectories of individual fish as well
as relationships between fish targets (e.g., proximity, nearest
neighbor angle, nipping/butting, circling, space utilization,
tortuosity, etc.).
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Spawning behavior.
(Click on the ""
button at the bottom left of the movie to start movie clip. Movie
may take several minutes to load. If QT is not installed on your
computer, go to http://www.apple.com/quicktime/download.html)
This QuickTime movie portrays a male fathead minnow under
a spawning structure that is joined by a gravid female. This movie demonstrates
several reproductive behaviors that are key to sucessful spawning
of this species including parallel circling and
a spawning attempt. The spawning attempt shows the male using
his entire body length to guide the female to deposit eggs to
underneath surface of the spawning structure. This behavior is
often repeated several times prior to egg deposition.
Other key behaviors, not shown in this clip, include pre-spawning
interactions (nipping & butting), male territoriality, egg
guarding and egg fanning. These behaviors are sufficiently distinctive
such that we can quantitatively discern them using our video
analysis software.
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Secondary Sex Characteristics
Panel A shows dorsal epithelial pad (arrow). Panel
B shows breeding tubercles on the snout (arrows). Panel
C shows ovipositor just anterior to anal fin; this is normal
for a breeding female but not normal for males. Ovipositors can
be observed on males exposed to low levels of estrogen-like compounds.
Since fathead minnows are sexually dimorphic (males and females
look notably different), and the male exhibits secondary sexual
characteristics in breeding condition, we can use changes in morphology of secondary sex characteristics as an ancillary
endpoint to evaluate exposure effects of low-level endocrine
disrupting chemicals. Expression of these secondary characteristics,
like reproductive behavior, is hormonally-driven. Thus, by integrating
behavior, hormones, and secondary sex characteristics, we can
obtain a more holistic picture of biologically-relevant responses
to endocrine disrupting chemicals.
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Related Links:
Kane, A.S., Salierno, J.D. and Brewer, S.K. 2004. Fish models in behavioral toxicology: automated
techniques, updates and perspectives. In Ostrander, G.K.,
ed. Methods in Aquatic Toxicology, Volume 2. Lewis Publishers,
Boca Raton, FL.
Kane, A.S., Salierno, J.D., Gipson, G.T., Molteno, T., and
, Hunter, C. 2004. Novel video-based movement analysis system to
quantify behavioral responses of fish. Water Research. 38:3993-4001.
Salierno, J.D., Gipson, G.T. and Kane, A.S. 2008. Quantitative movement analysis of social behavior
in mummichog, Fundulus heteroclitus. Journal of Ethology 26:35-42.
Lauer, L.E., McCarthy, M.M., Jong, J. and Kane, A.S. 2006. Sex differences in neuronal morphology in the
killifish hypothalamus. Brain Research, 1070:145-149.
Histological
Atlas of Normal Fathead Minnow Anatomy (coming soon)
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