Sampling technique to discriminate the different tissue layers of growing antler tips for gene discovery

Li, Chunyi; Clark, Dawn E.; Lord, E.A.; Stanton, Jo‐Ann L.; Suttie, J. M.

doi:10.1002/ar.10120

Cited by 99 publications

(97 citation statements)

References 10 publications

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“…The distal 5 cm of each antler main beam was used for this study. Tissue for mRNA extraction was rapidly separated into six morphologically defined layers (Li et al, 2002) under a dissecting microscope, snap-frozen in liquid nitrogen, and stored at À808C. Tissue for in situ hybridization and immunohistochemistry was fixed in 10% formalin for 12 hr and wax-embedded.…”

Section: Materials and Methods Tissue Collectionmentioning

confidence: 99%

“…The resulting library was thus enriched for antler-specific gene sequences. Details can be found in the paper by Li et al (2002). The clone used for in situ hybridization was CANS000A0044 and covers the sequence highlighted in Figure 8.…”

Section: Cloningmentioning

confidence: 99%

“…1). Antlerogenic stem cells have been found to be concentrated in the reserve mesenchyme underlying the dermis (Li et al, 2002(Li et al, , 2005a. Cells from the reserve mesenchyme divide and differentiate, thus contributing chondroblasts to the precartilage region, where progressive maturation occurs.…”

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confidence: 99%

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Expression of VEGF and pleiotrophin in deer antler

2006

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Deer antlers represent a unique model of mammalian regeneration in that they cast and fully regenerate every year. The deer antler thus provides a fascinating model of both rapid angiogenesis and chondrogenesis and the opportunity to investigate unique growth regulatory processes. One such phenomenon is the presence of vascularized cartilage in the growing antler tip-unlike other cartilage, which is typically avascular. The mechanisms by which blood vessels grow in the cartilage as well as the factors that drive antler extension at approximately 1 cm a day have been hitherto largely unknown. The aim of this study was to determine the expression of VEGF and pleiotrophin within the growing antler tip. We isolated cervine VEGF121 and VEGF165 from deer antler and found that mRNA is produced for VEGF in the precartilage and cartilage regions. By in situ hybridization, we examined whether the VEGF receptors Flt-1 and KDR are present in deer antler and found only KDR mRNA within the endothelial cells of the precartilage region. This finding is compatible with VEGF having an angiogenic effect within antler. Pleiotrophin mRNA was found in the vascular smooth muscle cells of the dermis, thus supporting a possible role in vascular growth. High levels of pleiotrophin mRNA were also detected in the precartilage region with possible implications for both angiogenesis and chondrogenesis. This is the first report of cervine angiogenic growth factors within the growing antler tip. Anat Rec Part A, 288A: 1281-1293, 2006. 2006 Key words: angiogenesis; cartilage; chondrogenesis; endothelial Systems where growth occurs at an astonishing rate such as the placenta, embryo, and deer antler can give important insights into the mechanisms involved in angiogenesis. Deer antlers are unique mammalian appendages in that they have an annual cycle of regeneration. During regeneration, the cartilage, skin, nerves, and blood vessels of the antler must grow at a rapid rate. This full regrowth occurs under conditions of low testosterone and high plasma IGF-1 during spring (Li et al., 2003). Antler grows at varying rates in different species, with growth rates reaching 12.5 mm/day in sika deer (Gao and Li, 1988), 27.5 mm/day in moose (Goss, 1970), and approximately 10 mm/day in red deer (Fennessy et al., 1991). Antler growth is driven from the tip, which consists of dermis, mesenchyme, precartilage, and the cartilage that transforms to bone and that then progressively calcifies within the antler (Fig. 1). Antlerogenic stem cells have been found to be concentrated in the reserve mesenchyme underlying the dermis (Li et al., 2002(Li et al., , 2005a. Cells from the reserve mesenchyme divide and differentiate, thus contributing chondroblasts to the precartilage region, where progressive maturation occurs. The cartilage zone contains columns of chondrocytes and large parallel vascular channels, which give a red appearance. In the region shown as the remodeling zone (Fig. 1) THE ANATOMICAL RECORD PART A 288A: 1281-1293 (2006) expand at a rapi...

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Section: Materials and Methods Tissue Collectionmentioning

confidence: 99%

Section: Cloningmentioning

confidence: 99%

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Expression of VEGF and pleiotrophin in deer antler

2006

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“…Price et al (2005a) used the term mesenchymal growth zone for the region of intense cell division (corresponding to Banks and Newbrey's reserve mesenchyme) present underneath the fibrous layer of the perichondrium. The zone proximally adjacent to the mesenchymal growth zone is referred to as the chondroprogenitor zone by Price et al (2005a), while Li et al (2002) use the term precartilage zone, which is also applied here (Figs. 3 and 4).…”

Section: Basic Aspects Of Antler Regeneration and Comparison With Limmentioning

confidence: 99%

Deer antler regeneration: Cells, concepts, and controversies

Kierdorf

Szuwart

2007

Journal of Morphology

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The periodic replacement of antlers is an exceptional regenerative process in mammals, which in general are unable to regenerate complete body appendages. Antler regeneration has traditionally been viewed as an epimorphic process closely resembling limb regeneration in urodele amphibians, and the terminology of the latter process has also been applied to antler regeneration. More recent studies, however, showed that, unlike urodele limb regeneration, antler regeneration does not involve cell dedifferentiation and the formation of a blastema from these dedifferentiated cells. Rather, these studies suggest that antler regeneration is a stem-cell-based process that depends on the periodic activation of, presumably neural-crest-derived, periosteal stem cells of the distal pedicle. The evidence for this hypothesis is reviewed and as a result, a new concept of antler regeneration as a process of stem-cell-based epimorphic regeneration is proposed that does not involve cell dedifferentiation or transdifferentiation. Antler regeneration illustrates that extensive appendage regeneration in a postnatal mammal can be achieved by a developmental process that differs in several fundamental aspects from limb regeneration in urodeles.

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“…These antlers were cleaned with 75% ethanol (v/v) carefully, removed by a surgical hand-saw, and then stored at À 80 1C after blood removal with a vacuum pump. The antler growing tips were collected according to the sampling technique described by Li et al [15]. Briefly, the incision was made surround the antler shaft approximately 1.3 cm proximal to the antler tip to collect the tip section, which is composed of skin, reserve mesenchyme and precartilage tissues.…”

Section: Sample Preparationmentioning

confidence: 99%