Altered beta-1,4-galactosyltransferase I expression during early inflammation after spinal cord contusion injury.
Post-traumatic inflammation has been implicated in secondary tissue damage after spinal cord injury (SCI). beta-1,4-Galactosyltransferase I (beta-1,4-GalT-I) is a key inflammatory mediator that plays a critical role in the initiation and maintenance of inflammatory reaction in diseases. The aim of the current study was to investigate whether beta-1,4-GalT-I is expressed in SCI. Spinal cord contusion model was established in adult rats. Real-time PCR and Western blot analysis were used to detect the spatio-temporal expression of beta-1,4-GalT-I after SCI. Lectin-fluorescent staining with RCA-I was used to detect the galactosylation of the membrane glycoproteins. The interaction and colocalization between beta-1,4-GalT-I and E-selectin in the injured spinal cords were also assessed by immunoprecipitation of E-selectin and double immunofluorescent staining, respectively. Real-time PCR revealed that beta-1,4-GalT-I mRNA reached the peak at 1d after spinal cord contusion. In situ hybridization indicated that beta-1,4-GalT-I mRNA was mainly distributed in the local inflammatory cells, adjacent to the center of injury. Double immunofluorescent staining showed that beta-1,4-GalT-I mostly overlapped with ED1-positive macrophages 1d after SCI, partly colocalized with microglia, neutrophils and a few with oligodendrocytes and astrocytes. The result of Lectin-fluorescent staining with RCA-I was similar to that of double immunofluorescent staining. Terminal galactosylation of E-selectin underwent obvious changes between sham and 3d after SCI by immunoprecipitation of E-selectin. Thus, the transient expression of high levels of beta-1,4-GalT-I may provide new insight into the early inflammation after SCI.
Niu
Shuqiong
S
The Jiangsu Province Key Laboratory of Neuroregeneration, Nantong University, Nantong, People's Republic of China.
Fei
Min
M
Cheng
Chun
C
Yan
Meijuan
M
Gao
Shangfeng
S
Chen
Mengling
M
Wang
Haibo
H
Li
Xin
X
Yu
Xiaowei
X
Qian
Ji
J
Qin
Jing
J
Zhao
Jian
J
Gu
Jianxin
J
Shen
Aiguo
A
D000818:Animals; D015153:Blotting, Western; D003288:Contusions / Q000150:complications / Q000201:enzymology*; D019040:E-Selectin / Q000096:biosynthesis / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005700:Galactosyltransferases / Q000096:biosynthesis* / Q000235:genetics; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D017403:In Situ Hybridization; D007249:Inflammation / Q000201:enzymology* / Q000209:etiology; D008297:Male; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics; D012313:RNA / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013119:Spinal Cord Injuries / Q000150:complications / Q000201:enzymology*
D019040:E-Selectin; D009419:Nerve Tissue Proteins; D012313:RNA; D005700:Galactosyltransferases; C119701:beta-1,4-galactosyltransferase I