Journal: BMC Gastroenterology

Article Title: Neuroprotective effects of vitamin D on high fat diet- and palmitic acid-induced enteric neuronal loss in mice

doi: 10.1186/s12876-018-0905-9

Figure Lengend Snippet: Overview of primary and secondary antibodies used in immunocytochemistry

Article Snippet: Stock solutions of palmitic acid (PA, P9767, Merck), VD (D1530, Merck), 7-dihydrocholesterol (7DHC), 16F16 (SML0021 Merck, PDIA3 inhibitor [ ]), GW69662 (M6191, Merck, peroxisome proliferator-activated receptor gamma (PPARγ) inhibitor [ ]), were prepared, aliquoted and stored at -20 °C.

Techniques: Purification

Journal: BMC Gastroenterology

Article Title: Neuroprotective effects of vitamin D on high fat diet- and palmitic acid-induced enteric neuronal loss in mice

doi: 10.1186/s12876-018-0905-9

Figure Lengend Snippet: Localisation of PDIA3 in mouse and primary cultures of myenteric neurons. Representative micrographs of ileum of normal diet 1x vitamin D animals ( a - c , g - i ) and of primary cultures myenteric neurons ( d - f , j - l ). a - f Immunocytochemical staining of neuronal cell bodies and fibres using PGP9.5 in ileum ( a ) and culture ( d ) and PDIA3 positive fibres and cell bodies in ileum ( b ) and culture ( e ), merged images showing PDIA3 is not co-located with enteric neurons in ileum ( c ) and culture ( f ). g - i Immunocytochemical staining to enteric glia using S100β in ileum ( g ) and culture ( j ) and PDIA3 positive fibres and cell bodies in ileum ( h ) and culture ( k ), merged images show co-localisation of S100β and PDIA3, suggesting PDIA3 is expressed in enteric glia. Bars represent 20μm

Article Snippet: Stock solutions of palmitic acid (PA, P9767, Merck), VD (D1530, Merck), 7-dihydrocholesterol (7DHC), 16F16 (SML0021 Merck, PDIA3 inhibitor [ ]), GW69662 (M6191, Merck, peroxisome proliferator-activated receptor gamma (PPARγ) inhibitor [ ]), were prepared, aliquoted and stored at -20 °C.

Techniques: Staining

Journal: BMC Gastroenterology

Article Title: Neuroprotective effects of vitamin D on high fat diet- and palmitic acid-induced enteric neuronal loss in mice

doi: 10.1186/s12876-018-0905-9

Figure Lengend Snippet: Effects of experimental treatment agent per se and on VD and VD+PA neuronal survival and PA-induced neuronal loss in primary cultures of myenteric neurons. a Supplementation with the protein disulphide isomerase family A member 3 (PDIA3) inhibitor 16F16 in the range 7x10 -8 M - 7x10 -7 M did not affect neuronal survival, but induced loss of all cells at 2x10 -5 M. b Supplementation with the PDIA3 inhibitor (10 -7 M) had no effect on the palmitic acid (PA, 4x10 -4 M), the 1α,25-hydroxy-vitamin D3 (VD, 10 -7 M) or the PA+VD effects on neuronal survival. c Peroxisome proliferator-activated receptor gamma (PPARγ) inhibitor, GW69662 (10 -6 M) did not prevent the PA-induced neuronal loss but prevented the VD-induced prevention of the PA-induced loss. d Supplementation with pre-immune sera (1:250) had no effects on PA-, VD- or PA+VD-induced effects on neuronal survival. e Supplementation of isocitrate lyase (ICL) immune sera (1:250) did not prevent the PA-induced loss but did prevent the protective effects of VD on PA-induced loss. Untreated controls were run in parallel. Data presented as mean ± SEM, control n =6-18 per treatment agent, PDIA3 inhibitor 16F16 n =6-12, PPARγ inhibitor, GW69662 n =6, pre-immune sera n =6, ICL immune sera n =6, PDIA3 inhibitor (10 -7 M) + PA/VD/VD+PA n =6, PPARγ inhibitor (10 -6 M) + PA/VD/VD+PA n =6-12, pre immune sera (1:250) n =12, ICL immune sera (1:250) n =12-18, ** p < 0.01, *** p <0.001

Article Snippet: Stock solutions of palmitic acid (PA, P9767, Merck), VD (D1530, Merck), 7-dihydrocholesterol (7DHC), 16F16 (SML0021 Merck, PDIA3 inhibitor [ ]), GW69662 (M6191, Merck, peroxisome proliferator-activated receptor gamma (PPARγ) inhibitor [ ]), were prepared, aliquoted and stored at -20 °C.

Techniques: