title: Distribution and expression of SLC45A2 in the skin of sheep with different coat colors.
abstract: BACKGROUND
  To investigate whether the membrane-associated transporter protein SLC45A2 is differentially expressed in the skin of sheep with different coat colors and to determine its correlation with coat color establishment in sheep.
  
  
  METHODS
  The expression of SLC45A2 in sheep skin samples with different coat colors was qualitatively and quantitatively analyzed by PCR amplification, RT-PCR, immunohistochemical staining and Western blotting.
  
  
  RESULTS
  A 193-bp SLC45A2 CDS sequence was successfully amplified from sheep skin samples with diverse coat colors. RT-PCR analysis revealed that SLC45A2 mRNA was expressed in all sheep skin samples tested, with relative expression levels of 512.74 ± 121.51 in black skin, 143.38 ± 119.31 and 1.36 ± 0.09 in black dots and white dots of piebald skin, respectively, and 1.02 ± 0.23 in white skin (p < 0.01**). Positive SLC45A2 protein bands were also detected in all skin samples by Western blot analysis, with relative expression levels of 0.85 ± ± 0.17** in black skin, 0.60 ± 0.05** and 0.34 ± 0.07 in black dots and white dots of piebald skin, respectively, and 0.20 ± 0.05 in white skin (p < 0.01**). Immunohistochemical assays revealed that SLC45A2 was expressed in the hair follicle matrix, the inner and outer root sheath, and the dermal papilla in the skin tissues with different coat colors. These patterns were quantified by optical density (OD) analysis, which yielded relative expression levels of 0.23 ± 0.11 in black skin, 0.19 ± 0.09 and 0.10 ± 0.03 in black dots and white dots of piebald skin, respectively, and 0.08 ± 0.01 in white skin (p < 0.05*).
  
  
  CONCLUSIONS
  SLC45A2 is detectably expressed in sheep skin of all coat colors, though at significantly different levels. SLC45A2 may participate in the establishment of coat color by regulating the synthesis and trafficking of melanin.
authors.forename: Haidong
authors.initials: H
authors.lastname: Wang
authors.forename: Linli
authors.initials: L
authors.lastname: Xue
authors.forename: Yanan
authors.initials: Y
authors.lastname: Li
authors.forename: Bingling
authors.initials: B
authors.lastname: Zhao
authors.forename: Tianzhi
authors.initials: T
authors.lastname: Chen
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Lucheng
authors.initials: L
authors.lastname: Chang
authors.affiliation: Department of Geriatrics, Tongji Hospital of Shanghai affiliated with Tongji University, Shanghai. whd1232123@163.com.
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D015870:Gene Expression; D006200:Hair Color / Q000502:physiology*; D018859:Hair Follicle / Q000166:cytology / Q000378:metabolism*; D007150:Immunohistochemistry; D026901:Membrane Transport Proteins / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012756:Sheep / Q000378:metabolism*; D034561:Sheep, Domestic; D012867:Skin / Q000378:metabolism; D012880:Skin Pigmentation / Q000502:physiology
chemical_list: D026901:Membrane Transport Proteins; D012333:RNA, Messenger
keywords: IHC; Q-PCR; SLC45A2; Western blot; coat color; sheep