Journal: PLoS Genetics

Article Title: Rgs6 is Required for Adult Maintenance of Dopaminergic Neurons in the Ventral Substantia Nigra

doi: 10.1371/journal.pgen.1004863

Figure Lengend Snippet: (A) Rgs6 expression revealed by immunohistofluorescence (red) together with TH (green) in mDA neurons of SNc but not VTA. Scale bar 410 µm. (B) Co-immunofluorescence analysis of TH (green) and Rgs6 or Pitx3 (red, as indicated) in SNc and VTA of adult mice. Arrowheads point to Pitx3-negative or Rgs6-negative mDA neurons in dorsal SNc. Scale bar 100 µm. (C) Triple immunofluorescence staining for TH (green), Pitx3 (red) and Calb1 (blue) on tissue sections of adult mice indicates that the majority of TH+Pitx3− cells of dSNc (arrowheads) are positive for Calb1, while TH+Pitx3+ cells of vSNc are negative for Calb1, consistent with depiction in A. Scale bar 100 µm.

Article Snippet: Primary antibodies used were against Pitx3 (rabbit home-made, 1∶400), Rgs6 (rabbit home-made 1∶25), Th (Millipore MAB318, 1∶1000), Th (Millipore AB152, 1∶500), Calb1 (R&D Systems AF3320, 1∶40), Fgf10 (Millipore ABN44, 1∶1000), Slc6a3 (Santa Cruz sc-32258, 1∶250), Drd2 (Santa Cruz sc5303, 1∶100), Aldh1a1 (Abcam ab24343, 1∶400), BDNF (Abcam ab108319, 1∶25), phospho-p27 (Abcam ab32096, 1∶50), phospho-Erk1/2 (Cell Signaling 4376, 1∶25), DJ-1 (Abcam ab18257, 1∶500) Lrrk2 (Epitomics 3514-1, 1∶50), Pink1 (Novus Biologicals BC100-494, 1∶50), LC3B (Cell Signaling 3868, 1∶50), and Vmat2 (Millipore AB1598P, 1∶200).

Techniques: Expressing, Immunohistofluorescence, Immunofluorescence, Staining

Journal: Endocrinology

Article Title: Exogenous 1,25-dihydroxyvitamin D3 exerts a skeletal anabolic effect and improves mineral ion homeostasis in mice that are homozygous for both the 1alpha-hydroxylase and parathyroid hormone null alleles.

doi: 10.1210/en.2006-0403

Figure Lengend Snippet: FIG. 1. Effect of exogenous 1,25(OH)2D3 on serum calcium and phosphorus, urine calcium relative to creatinine, and renal calcium transporters in vivo. Serum calcium (A) and phosphorus (B) and urine calcium to creatinine ratio (C) were determined in 2-wk-old vehicle-treated wild-type (WT-vehicle), vehicle-treated PTH/1(OH)ase/ mice (DKO-vehicle), and 1,25(OH)2D3-treated PTH/1(OH)ase/ mice (DKO1,25D) as described in Materials and Methods. Each value is the mean SEM of determinations in six mice of each group. D, Representative RT-PCR analysis of renal extracts for gene expression of TRPV5, calbindin-D9K (CB9K), calbindin-D28K (CB28K), and NCX1. Glyceraldehyde-3- phosphate dehydrogenase (GAPDH) mRNA was the loading control. E, Representative Western blots of renal extracts for expression of TRPV5, CB9K, CB28K, and NCX1. -Tubulin was used as a loading control for Western blots. F, TRPV5, CB28K, CB9,K and NCX1 mRNA levels relative to GAPDH mRNA level were assessed by densitometric analysis and expressed relative to respective levels of these transporters in wild-type mice. Each value is the mean SEM of triplicate determinations. G, TRPV5, CB28K, CB9K, and NCX1 protein levels relative to -tubulin protein level were assessed by densitometric analysis and expressed relative to respective levels of these transporters in wild-type mice. Each value is the mean SEM of triplicate determinations. *, P 0.05; ***, P 0.001, compared with vehicle-treated wild-type mice; #, P 0.05; ###, P 0.001, compared with vehicle-treated double-KO mice.

Article Snippet: Immunoblotting was carried out as described (24–26) using antibodies against TRPV5 (ECaC1), calbindin-D28K, and calbindin-D9K, Na /Ca2 exchanger 1 (NCX1) (Swant, Bellinzona, Switzerland) and -tubulin (Santa Cruz Biotechnology Inc., Santa Cruz, CA).

Techniques: In Vivo, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Control, Western Blot, Expressing