First Advisor

Deborah I. Lutterschmidt

Date of Publication

Winter 3-28-2019

Document Type


Degree Name

Master of Science (M.S.) in Biology






Red-sided garter snake -- Effect of cold on, Red-sided garter snake -- Reproduction, Enzymes, Biochemistry, Hibernation



Physical Description

1 online resource (v, 35 pages)


Environmental cues such as day length and temperature contribute to timing of biological rhythms in seasonal breeders. Life-history transitions such as spring emergence from hibernation, migration, or mating must be coordinated with environmental conditions or survival is compromised. Therefore, there must be chemical signaling pathways in the brain that transduce seasonally-changing sensory inputs into signals that initiate a hormonal cascade, culminating in reproductive behavior. The relative importance of environmental cues to reproductive timing varies with species, time of year, and sex, and the mechanisms driving these differences remain unknown. The role of photoperiod in regulating reproductive behavior has been explored in birds and mammals, but much less is known about the role of so-called supplementary cues such as temperature, which is crucial in the timing of ectotherm reproduction. This is a critical gap in our knowledge, because shifts in seasonal temperatures due to climate change could create a mismatch between peak reproductive behavior and resources necessary for gestation and offspring survival. Deiodinase 2 (DIO2) enzyme is a critical component of the pathway that mediates reproduction in photoperiod-activated seasonal breeders, but whether deiodinase 2 is sensitive to seasonal changes in environmental temperature is unknown. In this study, we used an ectothermic vertebrate known to be a temperature-activated seasonal breeder, the red-sided garter snake (Thamnophis sirtalis parietalis), to investigate changes in hypothalamic DIO2 in response to hibernation at 4°C and 12°C. We captured male and female snakes in Manitoba, Canada as they returned to their winter den site from summer feeding grounds. Snakes were hibernated in complete darkness at either 4°C or 12°C for up to 16 weeks. A subset of each sex and temperature group were euthanized at intervals, and the brains collected and processed for DIO2 immunohistochemistry. DIO2-specific staining was found in the anterior hypothalamus, in the periventricular hypothalamic nucleus and ventral pre-optic area, along the longitudinally central region of the olfactory tract, in the bed nucleus of the stria terminalis, caudally in the cortex and optic tectum, and in the lateral septal nucleus. DIO2-stained area in the anterior hypothalamus was quantified. Male T. sirtalis in both the 4°C and 12°C groups were found to have an increase in DIO2-specific staining in the anterior hypothalamus after 8 weeks in hibernation. Female T. sirtalis were found to have an increase in DIO2-specific staining in the anterior hypothalamus after 8 weeks in the 12°C group only. These findings shed light on the neuroendocrine pathway through which environmental cues other than photoperiod influence the timing of seasonal reproduction, and support the hypothesis that at least some components of this pathway are conserved across seasonal breeders.


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