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. 2016 Dec 19;5:e20914. doi: 10.7554/eLife.20914

Figure 5. Examining of Mg2+ balance in Trpm6-deficient adult mice.

(A–F) Assessment of 8 week-old Trpm6fl/+ (Control) and Trpm6Δ17/Δ17;Sox2-Cre (KO) littermate males. (A) Mg2+ levels in bones and (B) gastrocnemius muscle assessed by ICP-MS. (C) Immunostaining of kidney cryosections using a TRPM6-specific antibody. Representative images are shown (n = 2 tissues per genotype). The blue square indicates the position of the confocal and differential interference contrast magnified images acquired from control tissue. Arrows indicate labelling of the apical surface of renal tubules. (D) 24 hr urinary and (E) fecal Mg2+ excretion rates. (F) ISH on paraffin sections obtained from the colon of control and Trpm6-deficient mice (n = 2 tissues per genotype). (G–J) Examination of 6 month-old Trpm6fl/+ (Control) and Trpm6Δ17/fl;Ksp-Cre (Kidney KO) littermate males. (G) Immunostaining of TRPM6 in kidney cryosections. Arrows indicate labelling of renal tubules. (H–I) Determination of Mg2+ in serum (H) and bones (I). (J) 24 hr urinary Mg2+ excretion rate. (K–N) Assessment of 6 month-old Trpm6fl/+ (Control) and Trpm6Δ17/fl;Villin1-Cre (Intestine KO) littermate males. (K) ISH on paraffin sections of the colon using a Trpm6-specific probe (n = 2 tissues per genotype). (L, M) Mg2+ levels in the serum (L) and bones (M). (N) 24 hr urinary Mg2+ excretion rate. Data are represented as mean±SEM. ***-p≤0.001; **-p≤0.01; *-p≤0.05; n.s. – not significantly different (Student’s t-test); n – number of mice examined. Histological analysis in (F) and (K) was performed with n = 3 animals per group resulting in similar observations.

DOI: http://dx.doi.org/10.7554/eLife.20914.014

Figure 5.

Figure 5—figure supplement 1. Expression pattern of Trpm6 in the intestine.

Figure 5—figure supplement 1.

ISH on serial paraffin sections obtained from wildtype intestine using sense (left) and antisense (right) DIG-labelled probes for Trpm6. Boxes indicate the positions of the magnified images of the proximal and distal colon. Representative images of n = 3 tissues are shown. Arrows indicate Trpm6-positive cells in the colon. Note: Trpm6 transcripts were not detectable in the duodenum and ileum and specifically present in the absorptive epithelium cells of the proximal and distal colon.
DOI: http://dx.doi.org/10.7554/eLife.20914.015