A novel phenotypic marker for ATM-deficient 129S6/SvEvTac-ATMtm1Awb/J mice.

Ataxia-telangiectasia (A-T) is a human autosomal recessive disorder characterized by neuronal degeneration as well as many other physiological and somatic defects. ATM (A-T, mutated), the gene mutated in A-T, encodes a 370 kDa protein kinase. ATM knockout mouse models (ATM(-/-)) show growth retardation, infertility, neurological dysfunction, defects in T-lymphocytes, and extreme sensitivity to ionizing radiation. We have recently established multiple ATM(+/-) breeding pairs and discovered that all ATM(-/-) offspring exhibit a nonpigmented section of tail, usually at or near the tip. To our knowledge, this is the first time that a phenotype of nonpigmented tail has been reported in ATM(-/-) knockout mice. We believe that the sections of nonpigmented tail of 129S6/SvEvTac-ATM(tm1Awb)/J mice provide a novel phenotypic marker for research using this ATM knockout mouse model. Results from histochemistry and immunoblotting analysis further demonstrate that while melanocyte precursors or melanoblasts are present in the nonpigmented tail tissue of ATM(-/-) mice, they fail to differentiate fully into mature melanocytes. The potential connection between this phenotype and other clinical symptoms caused by ATM deficiency, such as progressive neuronal degeneration, is discussed in this article.

Vascular supplies differ in regenerating and nonregenerating amputated rodent digits.

Bone regenerates following amputation through the level of the nail, but bone is capped following amputation through more proximal levels. Because osteogenesis requires an ample blood supply, we postulated that a restricted vascular supply might be correlated with restricted regenerative ability at proximal levels. More than 40 rats and mice were injected with ink or resin to visualize vascular supplies of intact, regenerating, and nonregenerating rat and mouse digits. Ink-injected specimens were viewed as histological sections or cleared whole mounts. Partially digested resin casts were viewed using scanning electron microscopy. Contrary to our hypothesis, prior to amputation, proximal sites are more vascular than distal sites. At both proximal and distal levels, endosteal and periosteal vascular systems are evident. However, in proximal phalanges, additional subcutaneous and dermal layers encircle the bone. Beneath the distal nail, these layers are absent, and a single layer of vessels provides both periosteal and cutaneous supplies. After amputation at both levels, new vessels sprout profusely in osteogenic areas of both endosteum and periosteum. However, at proximal levels, the additional hypodermal and dermal vessels contribute to a vascular plexus that, paradoxically, may impair bone regrowth by contributing to the formation of dermal scar rather than bone.

Actin fiber patterns detected by Alexafluor 488 phalloidin suggest similar cell migration in regenerating and nonregenerating rodent toes.

Although mammals do not regenerate most appendages, they are able to regenerate toetips if the amputation occurs through the nail bed. The reasons for different outcomes following amputation at different levels are not understood. It is possible that cells at regenerating and nonregenerating sites migrate from fundamentally different tissues. If so, different migratory pathways could be detected. To identify putative migrating cells, microscope slides were made from both regenerating and nonregenerating toes of rats and mice on successive days after amputation. Fluorescent-labeled phalloidin, which binds polymerized f-actin, was used to identify actin filaments and fibers. Cells containing prominent actin bundles were distinguishable from those containing diffuse fibrils and those in which visible fibers were absent. Phalloidin labeling was similar in regenerating and nonregenerating digits after amputation. As early as 2 days after amputation at either proximal or distal levels, many cells of the hypodermis adjacent to the wound became labeled with phalloidin. The number and intensity of labeled hypodermal cells containing stress fiber-like bundles increased rapidly with time, and at successive times cells were seen progressively further distally. By approximately 7 days, they occupied the wound site immediately distal to bone of both regenerating and nonregenerating digits. Most dermal cells were unlabeled and endosteal and marrow cells contained only fibrillar actin. Phalloidin labeling does not support the concept of migration from different tissues in regenerating and nonregenerating amputation sites.