Epimorphic regeneration is the "holy grail" of regenerative medicine. Research aimed at investigating the various models of epimorphic regeneration is essential if a fundamental understanding of the factors underpinning this process are to be established. Deer antlers are the only mammalian appendages that are subject to an annual cycle of epimorphic regeneration. In our previous studies, we have reported that histogenesis of antler regeneration relies on cells resident within the pedicle periosteum (PP). The present study elaborates this finding by means of functional studies involving the deletion of PP. Four yearling and four 2-year-old stags were selected for total PP deletion or partial PP deletion experiments. Of the animals in the total PP deletion group, one showed no signs of antler regeneration throughout the antler growth season. Two showed substantial and one showed marginal delays in antler regeneration (at 34, 20 and 7 days, respectively) compared with the corresponding sham-operated sides. Histological investigation revealed that the delayed antlers were derived from regenerated PP. Unexpectedly, the regenerative capacity of the antler from the total periosteum-deleted pedicles depended on antler length at surgery. Of the four deer that had partial PP deletion, two regenerated antlers exclusively from the left-over PP on the pedicle shafts in the absence of participation from the pedicle bone proper. The combined results from the PP deletion experiments convincingly demonstrate that the cells of the PP are responsible for antler regeneration.

The process of angiogenesis is of interest because of the significant clinical benefits associated with controlling vascular growth. Within the antler, chondrogenesis and antler elongation are occurring at the rate of 1-2 cm per day and thus blood vessels are growing at this same rapid pace. We demonstrate that the process of angiogenesis in the antler is controlled at various tissue locations. The findings clearly differentiate the spatial location of the stem cells that drive chondrogenesis from the proliferation process driving the angiogenesis. Vessels within the lateral dermis contained BrdU-positive cells, suggesting that these vessels were elongating. Within the precartilage region, proliferating vessels were detected in bundles of complex structure evenly distributed throughout this tissue layer. The support cells within these bundles of vessels were detected by staining with a-smooth muscle actin, while the endothelial cells were negative. Additionally, the a-smooth muscle actin staining was found in association with the cartilage cells of the antler. The marked proliferation of the vascular associated cells in the precartilage region identified this area as a major region of vascular growth in the antler. We propose that within the precartilage region, the most likely mechanisms to explain the observed vascular morphology are that of vascular extension of the existing vessels and intussusceptive angiogenesis or sprouting to generate the small bundles of vessels. Anat Rec Part A, 288A: 973-981, 2006.2006 Wiley-Liss, Inc.

In a previous study (Li et al., 2001. J Exp Zool 290:18-30) a nude mouse model was established to investigate deer antler development. In that study we found nude mice could support the singularly implanted antlerogenic periosteum (AP) to form pedicle-like, but not antler-like, bony protuberances. To further develop the model and at the same time to use the updated model for the investigation of antler formation, three experiments were carried out in this study. The results showed that (1) antler-like protuberances were successfully induced on the nude mouse heads via subcutaneous co-transplantation of AP and deer skin, and subsequent exposure through wounding of the deer xenografts; (2) deer skin epidermis and its attached half thickness of dermis were sufficient to interact with the AP, and the interactions were capable of transforming adult scalp skin into velvet; (3) the putative initial inductive molecules were primarily derived from the AP cellular layer, rather than fibrous layer; (4) initiation of the ossification center in the avascular cartilage of each mouse "antler" took place via metaplasia, rather than classical endochondral ossification. Further research is required to identify means for effective stimulation of calcification of the "mouse antlers" in order to create the opportunity to investigate antler regeneration using the nude mouse model. Overall, the nude mouse model, once further developed, has the potential to become a powerful tool to study underlying mechanism of antlerogenesis and organogenesis/regeneration in general.

The aim of this study was to evaluate the effect of varying dietary P on bovine immune function. Nine first- or second-lactation Holstein cows were fed diets varying in P in a 3 x 3 Latin square design. Diets were formulated to contain either low (0.34%, no supplementary P), medium (0.43%), or high (0.52%) P. All 3 diets were formulated to meet or exceed current NRC requirements for P content. Between d 21 and 26 of each period, blood samples were collected and serum inorganic P concentration, lymphocyte proliferation, and neutrophil bactericidal activity were measured. Serum P increased with increasing dietary P intake and was greatest in the first lactation compared with subsequent lactations. There was a stage of lactation-dependent increase in lymphocyte proliferation after stimulation with concanavalin A, phytohemagglutinin, or pokeweed mitogen. However, dietary P did not alter lymphocyte proliferation or neutrophil bactericidal activity in vitro. In conclusion, decreasing dietary P to reduce manure P content and the risk of P losses from farms to surface water does not have an adverse effect on the innate or cell-mediated immune responses of lactating dairy cattle.

Deer antlers are the only mammalian appendage to display an annual cycle of full regeneration. The growth phase in antler involves the rapid proliferation of several tissues types, including epidermis, dermis, cartilage, bone, blood vessels, and nerves. Antlers thus provide an excellent model to study the developmental regulation of these tissues. We describe here the identification of two genes, pigment epithelium-derived factor (PEDF) and cyclin-dependent kinase inhibitor 1C (CDKN1C), both of which are known to be involved in cell proliferation and differentiation. These genes were identified as the result of screening an expressed sequence tag database derived from a cDNA library enriched for sequences from the growing antler tip. PEDF mRNA was detected in developing skin, cartilage, and bone during endochondral ossification. PEDF mRNA was not detected within endothelial cells that exhibited positive immunoreactivity to a CD146 antibody. CDKN1C mRNA was expressed by only the immature chondrocytes within the precartilage region. These results suggested that PEDF and CDKN1C are important genes involved in cell proliferation and differentiation during antler growth. Anat Rec 290: 994-1004, 2007. 2007 Wiley-Liss, Inc.