SECRETS OF CHEMORECEPTION: REGENERATION OF THE HARDERIAN GLAND KING YABUT DR. ROBERT T. MASON...
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Transcript of SECRETS OF CHEMORECEPTION: REGENERATION OF THE HARDERIAN GLAND KING YABUT DR. ROBERT T. MASON...
![Page 1: SECRETS OF CHEMORECEPTION: REGENERATION OF THE HARDERIAN GLAND KING YABUT DR. ROBERT T. MASON DEPARTMENT OF ZOOLOGY OREGON STATE UNIVERSITY.](https://reader036.fdocuments.net/reader036/viewer/2022070402/56649f255503460f94c3be40/html5/thumbnails/1.jpg)
SECRETS OF CHEMORECEPTION: REGENERATION OF THE HARDERIAN GLAND
KING YABUT
DR. ROBERT T. MASON
DEPARTMENT OF ZOOLOGY
OREGON STATE UNIVERSITY
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THE HARDERIAN GLAND
Discovered by Johann Jacob Harder in 1694
Found in virtually all land vertebrates
Function remains largely unknown
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THE HARDERIAN GLAND
Critical role in garter snake chemoreception Regeneration ability
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THAMNOPHIS SIRTALIS PARIETALISLIFE HISTORY
Warm summer months – feeding
Early fall – prepare for winter hibernation
Early spring – breeding
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THE VOMERONASAL SYSTEM AND THE ROLE OF SEMIOCHEMICALS Chemoreception is crucial to garter snake
behavior The VN system is used for detection of non-
volatile semiochemicals Female attractiveness sex pheromone Prey chemoattractants – earthworm shock
secretion proteinNC=nasal cavity
VNO=vomeronasal organ
HG=Harderian gland
LC=lacrimal canal
LD=lacrimal duct
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THE ROLE OF THE HARDERIAN GLAND
Female attractiveness sex pheromone is a lipid
VNO environment is aqueous Harderian Gland is the major source of fluid
for the VNO Dyes injected into the HG suggest link to VN
system NC=nasal cavity
VNO=vomeronasal organ
HG=Harderian gland
LC=lacrimal canal
LD=lacrimal duct
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THE ROLE OF THE HARDERIAN GLAND
Lipid binding proteins from Harderian Gland Homogenate of HG solubilized sexual
attractiveness pheromone Harderianectomized males do not court females
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PREVIOUS RESEARCH FROM MASON LAB
HDX snakes also show a loss in feeding behaviors
Chemosensory ability began to progressively return indicating a crucial role in feeding behavior
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PREVIOUS RESEARCH FROM MASON LAB
Study links HG to feeding behavior Previously unknown regeneration ability
indicates importance
Earthworm prey chemoattractant mechanism still unknown
http://www.crestock.com/image/476113-human-liver.aspx
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MY RESEARCH
Question: Does a relationship exist between the rate of Harderian Gland regeneration and chemosensory recovery in red-sided garter snakes?
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My Research
Hypotheis: The Harderian Gland plays a critical role in chemoreception of red-sided garter snakes and regeneration is correlated
to chemosensory ability.
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EXPERIMENTAL DESIGN
Purpose Establish a deficit in chemosensory ability using
behavioral and biochemical assays Track HG regeneration through time using
histology Treatments
HDX SHAM
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EXPERIMENTAL DESIGN
Two Protocols to establish deficit Latency to attack – behavioral assay Protein determination – biochemical assay
Photo by Don Powers
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LATENCY TO ATTACK PROTOCOL Collect earthworm shock secretion for swab
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Present swab to acclimated snake
LATENCY TO ATTACK PROTOCOL
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LATENCY TO ATTACK PROTOCOL
Time from first tongue flick until bite or to a max of 30 seconds (indicates no bite)
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RESULTS: SWAB TRIAL WEEK 1
Significant difference in bite times (P = 0.025)
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RESULTS: SWAB TRAIL WEEK 2
Non-biting SHAMs can be attributed to other factors
Recovered chemosensoryability of HDX?
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RESULTS: SWAB TRIAL WEEK 3
SHAM behavior fluctuates Consistent HDX non-biters
indicate deficit
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RESULTS: SWAB TRIAL WEEK 5
Treatment numbers dropdue to perfusions
Consistent non-biters remain
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RESULTS: SWAB TRIAL WEEK 8
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PERFUSIONS/HISTOLOGY
Perfusion performed weekly to preserve Harderian gland tissue for histology
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PROTEIN DETERMINATION
Quantify a difference in protein concentration of HG secretions between HDX and SHAM treatment groups
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HG SECRETION COLLECTION
Collect HG secretions Pilocarpine
Use secretioncollections to createdilutions with anunknown amount of protein
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PROTEIN DETERMINATION
Compare unknown dilutions to a standard curve of known protein concentrations to determine unknown concentrations
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RESULTS: PROTEIN DETERMINATION
SHAM HDX
Pro
tein
con
cent
ratio
n (
g/ l
)
0
2
4
6
8
P=0.027
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CONCLUSION
Behavioral assay Inconsistency in SHAM behavior may be due to
other factors Consistent HDX non-biters may indicate a deficit
in chemosensory ability Results are incomplete until correlated with
histology Protein determination
Significant statistical difference between SHAM and HDX protein concentrations (P = 0.027)
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ACKNOWLEDGEMENTS
Dr. Robert T. Mason Dr. Kevin Ahern and the Howard Hughes
Medical Institute Chris Friesen Rocky Parker Mason Lab –Ben Burke, Miranda Babcock-
Krenk, Mattie Squire, Kata Haeberlin