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Reduced white fat mass in adult mice bearing a truncated Patched 1.

Li Z, Zhang H, Denhard LA, Liu LH, Zhou H, Lan ZJ - Int. J. Biol. Sci. (2008)

Bottom Line: Hedgehog (Hh) signaling emerges as a potential pathway contributing to fat formation during postnatal development.Reduced total white fat mass and epididymal adipocyte cell size were observed in naturally occurring spontaneous mesenchymal dysplasia (mes) adult mice (Ptc1(mes/mes)), which carry a deletion of Ptc1 at the carboxyl-terminal cytoplasmic region.Taken together, our results indicate that deletion of carboxyl-terminal tail of Ptc1 can lead to the reduction of white fat mass during postnatal development.

View Article: PubMed Central - PubMed

Affiliation: Birth Defects Center, Department of Molecular, Cellular, Craniofacial Biology, University of Louisville Health Sciences Center, Louisville, KY 40202, USA.

ABSTRACT
Hedgehog (Hh) signaling emerges as a potential pathway contributing to fat formation during postnatal development. In this report, we found that Patched 1 (Ptc1), a negative regulator of Hh signaling, was expressed in the epididymal fat pad of adult mice. Reduced total white fat mass and epididymal adipocyte cell size were observed in naturally occurring spontaneous mesenchymal dysplasia (mes) adult mice (Ptc1(mes/mes)), which carry a deletion of Ptc1 at the carboxyl-terminal cytoplasmic region. Increased expression of truncated Ptc1, Ptc2 and Gli1, the indicators of ectopic activation of Hh signaling, was observed in epididymal fat pads of adult Ptc1(mes/mes) mice. In contrast, expression of peroxisome proliferator-activated receptor gamma, CCAAT/enhancer binding protein alpha, adipocyte P2 and adipsin were reduced in epididymal fat pads of adult Ptc1(mes/mes) mice. Taken together, our results indicate that deletion of carboxyl-terminal tail of Ptc1 can lead to the reduction of white fat mass during postnatal development.

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Increased expression of Ptc1, Ptc2, Gli1 and Gli2 and reduced expression of PPARgamma, CEBPalpha, aP2 and adipsin in epididymal WAT of Ptc1mes/mesmice. a. Representative RT-PCR analyses showing the loss of carboxyl-terminal Ptc1 mRNA (using Ptc1-F and Ptc1-R primers, 36 cycles) and increased expression of amino-terminal Ptc1 mRNA (using Ptc1-F2 and Ptc1-R2 primers, 32 cycles) in WAT of two 8-10-week-old Ptc1mes/mesmice. Deletion of 32 bp coding sequence within the exon 22 of Ptc1 in Ptc1mes/mes mice is highlighted by a yellow box. Two sets of RT-PCR primers for Ptc1 mRNA are indicated by arrows. b. Representative semi-quantitative RT-PCR showing the increased expression of amino-terminal Ptc1, Ptc2, Gli1 and Gli2 and the reduced expression of PPARgamma, CEBPalpha, aP2 and adipsin in epididymal WAT of 33-week-old Ptc1mes/mes mice. The numbers of PCR reaction cycles for carboxyl-terminal Ptc1 [Ptc1 (C): using primers Ptc1-F and Ptc1-R], amino-terminal Ptc1 [Ptc1-(N):using primers Ptc1-F2 and Ptc1-R2], Ptc2, Gli1, Gli2, Gli3, Gilz, CEBPalpha, PPARgamma, aP2, adipsin and beta-actin were 36, 38, 38, 26, 26, 38, 36, 28, 28, 20, 20 and 32, respectively. Experiments were repeated twice on independent RNA samples.
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Figure 3: Increased expression of Ptc1, Ptc2, Gli1 and Gli2 and reduced expression of PPARgamma, CEBPalpha, aP2 and adipsin in epididymal WAT of Ptc1mes/mesmice. a. Representative RT-PCR analyses showing the loss of carboxyl-terminal Ptc1 mRNA (using Ptc1-F and Ptc1-R primers, 36 cycles) and increased expression of amino-terminal Ptc1 mRNA (using Ptc1-F2 and Ptc1-R2 primers, 32 cycles) in WAT of two 8-10-week-old Ptc1mes/mesmice. Deletion of 32 bp coding sequence within the exon 22 of Ptc1 in Ptc1mes/mes mice is highlighted by a yellow box. Two sets of RT-PCR primers for Ptc1 mRNA are indicated by arrows. b. Representative semi-quantitative RT-PCR showing the increased expression of amino-terminal Ptc1, Ptc2, Gli1 and Gli2 and the reduced expression of PPARgamma, CEBPalpha, aP2 and adipsin in epididymal WAT of 33-week-old Ptc1mes/mes mice. The numbers of PCR reaction cycles for carboxyl-terminal Ptc1 [Ptc1 (C): using primers Ptc1-F and Ptc1-R], amino-terminal Ptc1 [Ptc1-(N):using primers Ptc1-F2 and Ptc1-R2], Ptc2, Gli1, Gli2, Gli3, Gilz, CEBPalpha, PPARgamma, aP2, adipsin and beta-actin were 36, 38, 38, 26, 26, 38, 36, 28, 28, 20, 20 and 32, respectively. Experiments were repeated twice on independent RNA samples.

Mentions: It has been reported that Ptc1mes mutant mice carry a deletion of a 32 bp DNA fragment in the exon 22 of Ptc1 allele 22 (Figure 3a). To confirm this, we isolated RNA from epididymal WAT of 8-10-week-old wild type and Ptc1mes/mes mice and performed RT-PCR analyses. As shown in Figure 3a, a positive DNA band covering exons 22 and 23 (using Ptc1-F and Ptc1-R primers) was detected in wild type, but not Ptc1mes/mesmice. These results confirmed the absence of the carboxyl-terminus of Ptc1 mRNA in epididymal WAT of Ptc1mes/mes mice. Since Ptc1 is a known downstream target gene of Hh signaling, RT-PCR analyses were also performed to determine whether expression of the amino region of the Ptc1 gene will be upregulated in epididymal WAT of Ptc1mes/mes mice. As shown Figure 3a, positive DNA bands covering exons 15 and 18 of Ptc1 (using Ptc1-F2 and Ptc1-R2 primers) were detected in wild type mice and even more pronounced in Ptc1mes/mes mice. Similar Ptc1 expression patterns were observed in epididymal WAT from 33-week-old mice (Figure 3b), indicating that the amino-terminal Ptc1 mRNA is overexpressed in WAT of Ptc1mes/mesmice.


Reduced white fat mass in adult mice bearing a truncated Patched 1.

Li Z, Zhang H, Denhard LA, Liu LH, Zhou H, Lan ZJ - Int. J. Biol. Sci. (2008)

Increased expression of Ptc1, Ptc2, Gli1 and Gli2 and reduced expression of PPARgamma, CEBPalpha, aP2 and adipsin in epididymal WAT of Ptc1mes/mesmice. a. Representative RT-PCR analyses showing the loss of carboxyl-terminal Ptc1 mRNA (using Ptc1-F and Ptc1-R primers, 36 cycles) and increased expression of amino-terminal Ptc1 mRNA (using Ptc1-F2 and Ptc1-R2 primers, 32 cycles) in WAT of two 8-10-week-old Ptc1mes/mesmice. Deletion of 32 bp coding sequence within the exon 22 of Ptc1 in Ptc1mes/mes mice is highlighted by a yellow box. Two sets of RT-PCR primers for Ptc1 mRNA are indicated by arrows. b. Representative semi-quantitative RT-PCR showing the increased expression of amino-terminal Ptc1, Ptc2, Gli1 and Gli2 and the reduced expression of PPARgamma, CEBPalpha, aP2 and adipsin in epididymal WAT of 33-week-old Ptc1mes/mes mice. The numbers of PCR reaction cycles for carboxyl-terminal Ptc1 [Ptc1 (C): using primers Ptc1-F and Ptc1-R], amino-terminal Ptc1 [Ptc1-(N):using primers Ptc1-F2 and Ptc1-R2], Ptc2, Gli1, Gli2, Gli3, Gilz, CEBPalpha, PPARgamma, aP2, adipsin and beta-actin were 36, 38, 38, 26, 26, 38, 36, 28, 28, 20, 20 and 32, respectively. Experiments were repeated twice on independent RNA samples.
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Figure 3: Increased expression of Ptc1, Ptc2, Gli1 and Gli2 and reduced expression of PPARgamma, CEBPalpha, aP2 and adipsin in epididymal WAT of Ptc1mes/mesmice. a. Representative RT-PCR analyses showing the loss of carboxyl-terminal Ptc1 mRNA (using Ptc1-F and Ptc1-R primers, 36 cycles) and increased expression of amino-terminal Ptc1 mRNA (using Ptc1-F2 and Ptc1-R2 primers, 32 cycles) in WAT of two 8-10-week-old Ptc1mes/mesmice. Deletion of 32 bp coding sequence within the exon 22 of Ptc1 in Ptc1mes/mes mice is highlighted by a yellow box. Two sets of RT-PCR primers for Ptc1 mRNA are indicated by arrows. b. Representative semi-quantitative RT-PCR showing the increased expression of amino-terminal Ptc1, Ptc2, Gli1 and Gli2 and the reduced expression of PPARgamma, CEBPalpha, aP2 and adipsin in epididymal WAT of 33-week-old Ptc1mes/mes mice. The numbers of PCR reaction cycles for carboxyl-terminal Ptc1 [Ptc1 (C): using primers Ptc1-F and Ptc1-R], amino-terminal Ptc1 [Ptc1-(N):using primers Ptc1-F2 and Ptc1-R2], Ptc2, Gli1, Gli2, Gli3, Gilz, CEBPalpha, PPARgamma, aP2, adipsin and beta-actin were 36, 38, 38, 26, 26, 38, 36, 28, 28, 20, 20 and 32, respectively. Experiments were repeated twice on independent RNA samples.
Mentions: It has been reported that Ptc1mes mutant mice carry a deletion of a 32 bp DNA fragment in the exon 22 of Ptc1 allele 22 (Figure 3a). To confirm this, we isolated RNA from epididymal WAT of 8-10-week-old wild type and Ptc1mes/mes mice and performed RT-PCR analyses. As shown in Figure 3a, a positive DNA band covering exons 22 and 23 (using Ptc1-F and Ptc1-R primers) was detected in wild type, but not Ptc1mes/mesmice. These results confirmed the absence of the carboxyl-terminus of Ptc1 mRNA in epididymal WAT of Ptc1mes/mes mice. Since Ptc1 is a known downstream target gene of Hh signaling, RT-PCR analyses were also performed to determine whether expression of the amino region of the Ptc1 gene will be upregulated in epididymal WAT of Ptc1mes/mes mice. As shown Figure 3a, positive DNA bands covering exons 15 and 18 of Ptc1 (using Ptc1-F2 and Ptc1-R2 primers) were detected in wild type mice and even more pronounced in Ptc1mes/mes mice. Similar Ptc1 expression patterns were observed in epididymal WAT from 33-week-old mice (Figure 3b), indicating that the amino-terminal Ptc1 mRNA is overexpressed in WAT of Ptc1mes/mesmice.

Bottom Line: Hedgehog (Hh) signaling emerges as a potential pathway contributing to fat formation during postnatal development.Reduced total white fat mass and epididymal adipocyte cell size were observed in naturally occurring spontaneous mesenchymal dysplasia (mes) adult mice (Ptc1(mes/mes)), which carry a deletion of Ptc1 at the carboxyl-terminal cytoplasmic region.Taken together, our results indicate that deletion of carboxyl-terminal tail of Ptc1 can lead to the reduction of white fat mass during postnatal development.

View Article: PubMed Central - PubMed

Affiliation: Birth Defects Center, Department of Molecular, Cellular, Craniofacial Biology, University of Louisville Health Sciences Center, Louisville, KY 40202, USA.

ABSTRACT
Hedgehog (Hh) signaling emerges as a potential pathway contributing to fat formation during postnatal development. In this report, we found that Patched 1 (Ptc1), a negative regulator of Hh signaling, was expressed in the epididymal fat pad of adult mice. Reduced total white fat mass and epididymal adipocyte cell size were observed in naturally occurring spontaneous mesenchymal dysplasia (mes) adult mice (Ptc1(mes/mes)), which carry a deletion of Ptc1 at the carboxyl-terminal cytoplasmic region. Increased expression of truncated Ptc1, Ptc2 and Gli1, the indicators of ectopic activation of Hh signaling, was observed in epididymal fat pads of adult Ptc1(mes/mes) mice. In contrast, expression of peroxisome proliferator-activated receptor gamma, CCAAT/enhancer binding protein alpha, adipocyte P2 and adipsin were reduced in epididymal fat pads of adult Ptc1(mes/mes) mice. Taken together, our results indicate that deletion of carboxyl-terminal tail of Ptc1 can lead to the reduction of white fat mass during postnatal development.

Show MeSH
Related in: MedlinePlus