Alterations in spinal cord gene expression after hindpaw formalin injection.
Heme oxygenase type 2 (HO-2) is an enzyme that uses heme as a substrate to produce iron, biliverdin, and carbon monoxide (CO). This enzyme participates in regulation of nociceptive signal transmission in spinal cord tissue. We set out to identify genes undergoing alterations in expression in a model of inflammatory pain and to determine whether HO-2 participates in that regulation. After the hindpaw injection of formalin in mice, we measured changes in expression of immediate early genes including c-fos, c-jun, jun B, nerve growth factor induced genes (NGFI-A and NGFI-B) and activity-related cytoskeletal protein (ARC) using real-time PCR. The mRNA corresponding to these genes increased in abundance in the first hour after formalin injection and then slowly declined. Changes in the abundance of prodynorphin, extracellular signal related kinases (ERK1 and ERK2) and N-methyl-D-aspartate (NMDA) receptor R1 subunit mRNA generally peaked between 8 and 12 hr after formalin injection. In HO-2 null mutant mice, the enhancement of expression was less for all genes studied. We went on to quantify gene expression in superficial dorsal horn tissue using laser capture microdissection followed by RNA amplification and real-time PCR. The results confirmed that the changes in gene expression were occurring in regions of the spinal cord involved in nociceptive processing. We conclude that the hindpaw injection of formalin leads to enhanced early and late expression of many genes in spinal cord dorsal horn tissue, and that this enhancement of expression relies to a degree on the presence of HO-2.
Li
Xiangqi
X
Department of Anesthesiology, Stanford University, Palo Alto, California 94304, USA.
Lighthall
Geoff
G
Liang
De-Yong
DY
Clark
J David
JD
D000704:Analysis of Variance; D000818:Animals; D001522:Behavior, Animal; D004195:Disease Models, Animal; D005557:Formaldehyde / Q000633:toxicity*; D007839:Functional Laterality; D015870:Gene Expression / Q000187:drug effects* / Q000502:physiology; D006419:Heme Oxygenase (Decyclizing) / Q000172:deficiency / Q000235:genetics; D006614:Hindlimb / Q000187:drug effects; D007150:Immunohistochemistry / Q000379:methods; D007249:Inflammation / Q000139:chemically induced / Q000150:complications / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D042282:Microdissection / Q000379:methods; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D010146:Pain / Q000209:etiology / Q000378:metabolism*; D011519:Proto-Oncogenes / Q000235:genetics / Q000502:physiology; D012333:RNA, Messenger / Q000096:biosynthesis; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013116:Spinal Cord / Q000187:drug effects* / Q000378:metabolism; D013997:Time Factors
D012333:RNA, Messenger; D012097:Repressor Proteins; D005557:Formaldehyde; D006419:Heme Oxygenase (Decyclizing); C105245:heme oxygenase-2; D020928:Mitogen-Activated Protein Kinases