シマヘビの器官の位置を研究したLASSOのデータの実際の値

Description

Table S1: The results of LASSO linear regression based on distances from the snout to each organ. Table S2: The results of LASSO linear regression based on vertebral counts from the first cervical.

Steps to reproduce

Twenty adult female E. quadrivirgata were obtained from the Japan Snake Institute (Ohta, Japan). Following Matsubara et al. (2014), individuals were euthanized after incision of the trunk. The ovaries and uterus were removed, and marker pins were inserted into the centers of the heart and gallbladder to mark corresponding vertebral positions. Because the liver and kidneys are elongated, pins were inserted at both anterior and posterior ends of each organ. Based on pin positions, 1 mm diameter copper wire was looped around the vertebrae located dorsally to each marked point and fixed in place. Internal organs were then removed. Because the copper wire remains after skeletal staining, it allowed identification of vertebral positions corresponding to each organ. Samples were skinned and fixed in 70% ethanol for 5 days. All staining procedures were conducted at room temperature. Samples were then transferred to 99.5% ethanol for 3 days, followed by 100% acetone for 3 days. After removal of acetone, samples were transferred to ion-exchanged water overnight. The following day, samples were immersed in 1% KOH for 3 hours and then stained in 0.005% Alizarin Red in 1% KOH for 10 days to visualize ossified tissue. After staining, samples were cleared in 1% KOH for 3 days, followed by treatment in 0.7% KOH with 30% glycerol for 2 days, and then 0.3% KOH with 70% glycerol for 2 weeks. Finally, samples were cleared in 100% glycerol for 1 month. After clearing, samples were removed from glycerol, and distances from the snout to each copper wire position were measured. Vertebral counts from the first cervical vertebra to each wire position were also recorded. These measurements provided organ positions in both length and vertebral units. Pearson’s correlation coefficients were calculated among all organ positions, and statistical significance was adjusted for multiple comparisons using the Benjamini–Hochberg procedure. LASSO regression was applied to evaluate explanatory relationships among organ positions: z_i ≈ ∑_(j≠i)▒〖w_ij z_j 〗, where z_i denotes the z-score-standardized position of an organ, and w_ij denotes the regression coefficient representing the contribution of z_j to the prediction of z_i.

Institutions

• Osaka Metropolitan University

  Osaka, Osaka

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