Computational protocol: Seasonal morphometry of the vomeronasal organ in the marsupial mouse, Antechinus subtropicus

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Protocol publication

[…] The data generated were used to investigate the following research questions: Is body size influenced by breeding condition?Are there differences between the size of the left and right parts of the VNO?Is the size of the VNO influenced by body size?Is the size of the VNO influenced by breeding condition when adjusted for any effects of body size?Is the amount of sensory tissue contained in the VNO influenced by body size?Is the amount of sensory tissue contained in the VNO influenced by breeding condition when adjusted for any effects of body size? Is body size influenced by breeding condition?Are there differences between the size of the left and right parts of the VNO?Is the size of the VNO influenced by body size?Is the size of the VNO influenced by breeding condition when adjusted for any effects of body size?Is the amount of sensory tissue contained in the VNO influenced by body size?Is the amount of sensory tissue contained in the VNO influenced by breeding condition when adjusted for any effects of body size?Statistical analysis was performed using SPSS Version 22 (Armonk, NY: IBM Corp), figures were drawn using the R package ggplot2 version 2.1.0. The statistical analysis is described in detail in the Supporting Information, and summarized here: Tests of normality (Skewness and Kurtosis) of the dependent variables associated with body size were carried out on the whole population, the population divided by sex and on the individual groups. Homogeneity of variance (Levene's test) was tested on the groups of the male and female populations. The majority of the parameters showed a normal distribution and homogeneity of variance for all breeding groups but because of a few exceptions and also because the small group sizes (n = 6) limited the power of these tests to detect non‐normality and heterogeneity of variance, corrections for heterogeneity of variance (Welch's correct) and post hoc testing which did not assume equal variance (Games‐Howell), were used in the analysis of variance for these parameters. Possible asymmetry of the VNO and associated structures was investigated using paired comparisons (paired t test, left vs. right) and when it was found that no biologically significant asymmetry existed, data was pooled for further investigation. For VNO metrics involving length or height, the mean of left and right were used, for metrics involving areas of volumes, left and right were summed.The relationship between the length of the VNO and the size of the animal was then investigated using multiple linear regression with length of VNO as dependent variable and the body‐size related parameters as predictor variables.Finally, since a relationship between body size and VNO was demonstrated, the differences in VNO metrics between breeding groups were explored in male and female animals using analysis of covariance (ANCOVA) with the best predictor of VNO length as the covariate.Because this analysis involved multiple dependent variables, Benjamini–Hochberg correction was used to maintain an overall 5% false discovery rate. In addition, because of the small group sizes, which had limited power to detect non‐normality and heterogeneity of variance, the analyses were carried out with 1,000 sample bootstrapping generating bias‐corrected accelerated confidence intervals. In all cases, bootstrapping produced results which supported the initial analysis. All of the figures and tables show the results of the initial analyses. […]

Pipeline specifications

Software tools SPSS, Ggplot2
Application Miscellaneous
Organisms Mus musculus, Homo sapiens