Validation of salivary oxytocin and vasopressin as biomarkers in domestic dogs

MacLean, Evan L.; Gesquiere, Laurence R.; Gee, Nancy R.; Levy, Kerinne; Martin, W.H.; Carter, C. Sue

doi:10.1016/j.jneumeth.2017.08.033

Cited by 94 publications

(98 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Finally, the dogs were led back to their crates for an additional 15 min, when the last samples were collected (T2), in accordance with the timings previously reported for determining salivary cortisol concentrations in dogs after exposure to a stressor [43,44]. All samples were immediately refrigerated and brought to the Etovet laboratory of the Department of Veterinary Sciences-University of Pisa (Italy) for centrifugation and stocking at −20 • C, until they were analyzed for oxytocin quantification with a Cayman Chemical ELISA Kit ® (Item #500440) (Ann Arbor, MI, USA), previously validated for use in dogs [33], and cortisol quantification by Diametra Cortisol Enzyme Immunoassay Kit ® (Spello, PG, Italy), according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

“…Lastly, salivary OXT sampling has none of these disadvantages; it is a noninvasive procedure, it can be performed with specific and accurate timing (although the right timing of OXT peak in saliva has not yet been accurately established), and it can be done directly by owners in the dogs' usual environment. Furthermore, salivary EIA measurement can also be carried out without a solid-phase extraction [33].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Effects of Stroking on Salivary Oxytocin and Cortisol in Guide Dogs: Preliminary Results

Ogi

Mariti

Baragli

et al. 2020

Animals

View full text Add to dashboard Cite

This pilot study aimed at investigating how salivary oxytocin levels are affected by human interaction and isolation in eight guide dogs (six Labrador retrievers and two golden retrievers; four males and four females, 21.87 ± 1.36 months old) just before assignment to the blind person. Each dog engaged, at one-week intervals, in a positive (5 min of affiliative interaction with their trainer) and a negative (5 min of isolation) condition. Saliva samples used for Enzyme Immunoassay (EIA) quantification of salivary oxytocin were collected before and immediately after both experimental conditions. In order to assess potential hypothalamic pituitary adrenal (HPA) axis activation that could have affected oxytocin levels, saliva samples were collected 15 min after both experimental conditions for EIA quantification of salivary cortisol and a behavioral assessment was performed during the negative condition. The results were compared using the Wilcoxon test (p < 0.05). Oxytocin concentrations showed a statistically significant increase after the positive interaction (p = 0.036) and no difference after the negative one (p = 0.779). Moreover, no difference (p = 0.263) was found between the cortisol concentrations after each experimental condition and no signs of distress were observed during the isolation phase. These preliminary findings support the hypothesis that stroking dogs has positive effects on their emotional state independently of hypothalamic pituitary adrenal axis activation.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effects of Stroking on Salivary Oxytocin and Cortisol in Guide Dogs: Preliminary Results

Ogi

Mariti

Baragli

et al. 2020

Animals

View full text Add to dashboard Cite

show abstract

“…To assess the levels of relevant hormones, we examined their concentrations in saliva. A recent study by MacLean [31] validated salivary measures of OT and AVP in dogs. Because saliva collection is simple and painless, it is better suited for assessments in dogs than other methods, such as using plasma or urine, which present a host of challenges related to design, analysis, and welfare [31].…”

Section: Introductionmentioning

confidence: 99%

Salivary Vasopressin as a Potential Non–Invasive Biomarker of Anxiety in Dogs Diagnosed with Separation–Related Problems

Pirrone

Pierantoni²,

Bossetti

et al. 2019

Animals

View full text Add to dashboard Cite

Physiological biomarkers of canine anxiety have not been extensively investigated to date. To identify new biomarkers in dogs, we compared behaviorally normal dogs (Control group, N = 13) to dogs diagnosed with separation problems (Case group, N = 13) as they were introduced into a novel environment in the presence of two strangers and subjected to a short episode of separation and reunion with the owner. During the separation phase, dogs in the Case group explored significantly less than controls and were significantly more persistent in expressing passive stress-coping strategies aimed at seeking proximity to their owners. When the owners returned, dogs with separation distress spent significantly more time jumping up on the strangers than control dogs did. Salivary oxytocin and vasopressin concentrations did not differ between samples taken before and after the separation. However, vasopressin concentrations immediately after separation were significantly higher in the Case than in the Control group and remained higher, although not significantly so, 10 min later. These results indicated that dogs with separation distress became more anxious than typical dogs when separated from their owner in an unfamiliar environment and provided preliminary support for the use of salivary vasopressin as a possible biomarker for anxiety-related responses in dogs.

show abstract

“…61‐6520, Thermo Fisher, Waltham, MA, USA) was added and incubated for 2 hours. After rinsing, the plate was incubated for 10 minutes at 26°C with ELISA‐peroxidase substrate 3,3′,5,5′‐tetramethylbenzidine (100 µL well ‐1 ; Nacalai Tesque, Kyoto, Japan), as described previously . Absorbance was measured at 450 nm using a 680 microplate reader (Bio‐Rad).…”

Section: Methodsmentioning

confidence: 99%

A monoclonal antibody raised against a synthetic oxytocin peptide stains mouse hypothalamic neurones

Kogami

Tsuji

et al. 2020

J Neuroendocrinology

View full text Add to dashboard Cite

A monoclonal antibody against oxytocin was generated in 7a5 hybridoma cells derived from myeloma cells and lymphocytes from the spleen of mice immunised with a synthetic oxytocin peptide. The 7a5 monoclonal antibody bound with oxytocin in enzyme‐linked immunosorbent assays. 7a5 cell growth medium was diluted up to 5000‐fold and used for immunohistochemistry. First, to test the specificity of the 7a5 antibody against oxytocin, we stained brain tissues of oxytocin knockout mice, comprising mice in which the first exon of the oxytocin‐neurophysin gene is deleted. No 7a5 immunoreactivity was detected in the paraventricular nucleus (PVN) of the hypothalamus of oxytocin knockout mice; however, this area was strongly stained with the anti‐vasopressin polyclonal antibody, HM07. Tissue preparations of the wild‐type mouse PVN and supraoptic nucleus (SON) displayed 7a5 immunoreactivity that was indistinguishable from the staining produced with an anti‐oxytocin polyclonal antibody, HM06. The immunoreactivity of HM06 in the PVN was similar to that of an anti‐oxytocin monoclonal antibody, PS38. We then examined the cross‐reactivity of 7a5 with arginine vasopressin. The majority of cell soma and processes stained by 7a5 were not co‐stained with the vasopressin antibody in SON and PVN regions. Furthermore, the suprachiasmatic nucleus was stained by the vasopressin antibody but not by 7a5. These results demonstrate that 7a5 is a new anti‐oxytocin monoclonal antibody recognising oxytocin and not vasopressin; therefore, 7a5 can be used to investigate the role of oxytocin in the brain.

show abstract

Validation of salivary oxytocin and vasopressin as biomarkers in domestic dogs

Abstract: OT and AVP are present in dog saliva, and ELISAs for their detection are methodologically valid.

Cited by 94 publications

References 49 publications

Effects of Stroking on Salivary Oxytocin and Cortisol in Guide Dogs: Preliminary Results

Effects of Stroking on Salivary Oxytocin and Cortisol in Guide Dogs: Preliminary Results

Salivary Vasopressin as a Potential Non–Invasive Biomarker of Anxiety in Dogs Diagnosed with Separation–Related Problems

A monoclonal antibody raised against a synthetic oxytocin peptide stains mouse hypothalamic neurones

Contact Info

Product

Resources

About