Nerve growth factor (NGF) is synthesized in cutaneous wound tissue, and its higher levels in the neonate may contribute to more efficient wound healing. We used in situ hybridization and immunohistochemistry to define NGF mRNA and protein expression in intact skin and following excision wounding in neonatal and adult rats. To determine whether NGF is associated with wound contractile fibroblasts (myofibroblasts), we also examined expression of alpha-smooth muscle actin (alpha-SMA) mRNA and protein, established markers for these cells. In intact skin, NGF mRNA and protein were present in vascular and arrector pili smooth muscle, hair follicle sheath cells, keratinocytes, and hypodermal fibroblasts. Neonatal adipocytes and Schwann cells also expressed NGF mRNA and protein, while adult adipocytes and Schwann cells displayed only NGF-ir. Following wounding, NGF mRNA expression was exuberant in these cell types, and increased similarly at both ages and appeared de novo in skeletal muscle cells. Additionally, both NGF mRNA and protein were present in macrophages and myofibroblasts, and expression in myofibroblasts was significantly greater in neonates. Wound myofibroblasts also expressed alpha-SMA. Surprisingly, after wounding alpha-SMA mRNA and protein were present in essentially all cells in which NGF mRNA was detected. We conclude that NGF expression is enhanced in many cell types after wounding, but greater NGF synthesis in neonates appears to be due to a more robust myofibroblast response. In addition, cell types which demonstrated NGF mRNA also expressed alpha-SMA, and staining for both markers increased following wounding, suggesting synthesis of both proteins is regulated in a coordinated fashion.

The periorbital sheath serves as a major pathway for sympathetic nerves traveling to distal orbital targets in the rat. This tissue accommodates sympathetic fiber sprouting in the neonate but becomes impassable by postnatal day 30 (PND 30). In contrast, smooth muscle target remains receptive to sympathetic ingrowth. To determine the attributes of receptive and nonreceptive tissues, we compared periorbital pathway and target tissue phenotypes prior to (PND 5 and PND 15) and after (PND 30 and PND 60) the period when pathway receptivity is lost. Both pathway cells and superior tarsal smooth muscle cells expressed α‐smooth muscle actin and smooth muscle myosin heavy chain throughout development. At PND 5–15, both tissues also expressed vimentin, collagen IV, laminin 1 and laminin β2, whereas fibronectin was detected only in pathway tissue. At PND 30, vimentin, collagen IV, and fibronectin were absent in tarsal muscle but were robust in pathway tissue. Laminin 1 and laminin β2 expression was maintained in muscle; however, in pathway cells, laminin 1 declined modestly, and laminin β2 decreased precipitously to barely detectable levels. Quantitative competitive polymerase chain reaction showed that nerve growth factor mRNA was present in the pathway throughout development at levels that were greater than both surrounding connective tissue and tarsal muscle. We conclude that the loss of pathway receptivity to sympathetic nerve ingrowth is associated with a transition from a phenotype similar to fetal smooth muscle cells to one that is more consistent with myofibroblast‐like cells. J. Comp. Neurol. 408:352–364, 1999. © 1999 Wiley‐Liss, Inc.