Evan L MacLean1, Laurence R Gesquiere2, Nancy Gee3, Kerinne Levy4, W Lance Martin5, C Sue Carter6. 1. School of Anthropology, University of Arizona, Tucson, AZ 85721, United States. Electronic address: evanmaclean@arizona.edu. 2. Department of Biology, Duke University, Durham, NC 27708, United States. 3. Waltham Centre for Pet Nutrition, Waltham on the Wolds, Leicestershire, LE14 4RT, United Kingdom; Department of Psychology, State University of New York, Fredonia, NY 14063, United States. 4. Canine Companions for Independence, Santa Rosa, CA, 95407, United States. 5. Martin-Protean LLC, Princeton, NJ 08540, United States. 6. Kinsey Institute and Department of Biology, Indiana University, Bloomington, IN 47405, United States.
Abstract
BACKGROUND: Oxytocin (OT) and Vasopressin (AVP) are phylogenetically conserved neuropeptides with effects on social behavior, cognition and stress responses. Although OT and AVP are most commonly measured in blood, urine and cerebrospinal fluid (CSF), these approaches present an array of challenges including concerns related to the invasiveness of sample collection, the potential for matrix interference in immunoassays, and whether samples can be collected at precise time points to assess event-linked endocrine responses. NEW METHOD: We validated enzyme-linked immunosorbent assays (ELISAs) for the measurement of salivary OT and AVP in domestic dogs. RESULTS: Both OT and AVP were present in dog saliva and detectable by ELISA and high performance liquid chromatography - mass spectrometry (HPLC-MS). OT concentrations in dog saliva were much higher than those typically detected in humans. OT concentrations in the same samples analyzed with and without sample extraction were highly correlated, but this was not true for AVP. ELISA validation studies revealed good accuracy and parallelism, both with and without solid phase extraction. Collection of salivary samples with different synthetic swabs, or following salivary stimulation or the consumption of food led to variance in results. However, samples collected from the same dogs using different techniques tended to be positively correlated. We detected concurrent elevations in salivary and plasma OT during nursing. COMPARISON WITH EXISTING METHODS: There are currently no other validated methods for measuring OT/AVP in dog saliva. CONCLUSIONS: OT and AVP are present in dog saliva, and ELISAs for their detection are methodologically valid.
BACKGROUND: Oxytocin (OT) and Vasopressin (AVP) are phylogenetically conserved neuropeptides with effects on social behavior, cognition and stress responses. Although OT and AVP are most commonly measured in blood, urine and cerebrospinal fluid (CSF), these approaches present an array of challenges including concerns related to the invasiveness of sample collection, the potential for matrix interference in immunoassays, and whether samples can be collected at precise time points to assess event-linked endocrine responses. NEW METHOD: We validated enzyme-linked immunosorbent assays (ELISAs) for the measurement of salivary OT and AVP in domestic dogs. RESULTS: Both OT and AVP were present in dog saliva and detectable by ELISA and high performance liquid chromatography - mass spectrometry (HPLC-MS). OT concentrations in dog saliva were much higher than those typically detected in humans. OT concentrations in the same samples analyzed with and without sample extraction were highly correlated, but this was not true for AVP. ELISA validation studies revealed good accuracy and parallelism, both with and without solid phase extraction. Collection of salivary samples with different synthetic swabs, or following salivary stimulation or the consumption of food led to variance in results. However, samples collected from the same dogs using different techniques tended to be positively correlated. We detected concurrent elevations in salivary and plasma OT during nursing. COMPARISON WITH EXISTING METHODS: There are currently no other validated methods for measuring OT/AVP in dog saliva. CONCLUSIONS: OT and AVP are present in dog saliva, and ELISAs for their detection are methodologically valid.
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