• Animal cells and systems
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• 제16권6호
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• pp.462-468
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• 2012

Urodeles including newt and salamander have remarkable regenerative capacity during the postembryonic life span. Some of the unique features are the formation of the well-developed wound epidermis and the active dedifferentiation process in the early phase of regeneration. These are regarded as key events for the successful regeneration since no further regenerative activity is possible without them. In this study, we investigated the role of retinoic acid (RA) in salamander limb regeneration by blocking RA synthesis using disulfiram, an inhibitor of aldehyde dehydrogenase that oxidizes retinal to RA. Disulfiram treatment resulted in delaying the limb regeneration processes via inhibition of wound epidermis formation and dedifferentiation process. When RA was administered after disulfiram treatment, the inhibitory effect of disulfiram was rescued. In addition, disulfiram treatment after the dedifferentiation stage resulted in the mild retardation of limb regeneration, suggesting that RA might also be involved in the blastema outgrowth. Furthermore, salamander MMP-9 gene expression was also inhibited by disulfiram treatment. Collectively, our findings indicate that endogenous RA may play an important role(s) in the early phase of limb regeneration by regulating the expression of molecules responsible for the modification of intracellular and extracellular environment during salamander limb regeneration.

• Animal cells and systems
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• 제14권2호
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• pp.73-81
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• 2010

In this study, monoclonal antibodies against lysosomal acid phosphatase (LAP) of a salamander, Hynobius leechii, were used to determine the spatial and temporal expression of the LAP in the regenerating limbs. The Western blot and immunohistochemical analysis in the limb regeneration revealed that LAP was highly expressed at the dedifferentiation stage, especially in the wound epidermis and dedifferentiating limb tissues such as muscle and cartilage. With RA treatment, the LAP expression became upregulated in terms of both level and duration in the wound epidermis, blastemal cell and dedifferentiating limb tissues. In addition, in situ activity staining of LAP showed a similar result to that of immunohistochemistry. Thus, the activity profile of LAP activity coincides well with the expression profile of LAP during the dedifferentiation period. Furthermore, to examine the effects of lysosomal enzymes including LAP on salamander limb regeneration, lysosome extract was microinjected into limb regenerates. Interestingly, when the lysosome extract was microinjected into limb regenerates with a low dose of RA($50\;{\mu}g/g$ body wt.), skeletal pattern duplication occurred frequently in the proximodistal and transverse axes. Therefore, lysosomal enzymes might cause the regenerative environment and RA plays dual roles in the modification of positional value as well as evocation of extensive dedifferentiation for pattern duplication. In conclusion, these results support the hypothesis that dedifferentiation is a crucial event in the process of limb regeneration and RA-evoked pattern duplication, and lysosomal enzymes may play important role(s) in this process.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 1998년도 제4차 학술발표대회 및 정기총회
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• pp.57-58
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• 1998

From the present study, following conclusions can be drawn: 1. lide in other species, axolotl FGF-8 is proposed to play a similar role in the early phase of limb development. However, the mechanism of its expression might be somewhat different from amniotes considering its characteristic mesenchymal expression. 2. In the regenerating axolotl limbs, Fgf-8 expression profile suggests that it is involved in wound gealing, dedifferentiation, and blastema formation. 3. Exoggenously supplied FGF-8 can accelerate blastema formation and concomitantly increase the Msx-1 expression level at the early stage of limb regeneration. Furthermore, it can partially substitute for nerve factor(s) as has been indicated by the induction of blastema formation in the denervated regenerates after FGF-8 application. 4. The unique expression feature of Fgf-8 in hte mesenechymal tissue of the regenerating axolotl limb might be casually related to its remarkable regeneration capacity of urodele.

• Animal cells and systems
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• 제4권2호
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• pp.145-150
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• 2000

Retinoic acid (RA) evokes pattern duplication in the regenerating salamander limb. Interestingly, it also enhances dedifferentiation in the regenerate by the morphological, histological and biochemical criteria. To examine whether there is any correlation between the RA-evoked pattern duplication and de novo protein synthetic profile in the regenerating salamander limb, especially during dedifferentiation, we analyzed stage-specific protein synthesis pattern in the normal and RA-treated regenerating limbs by metabolic labeling followed by two-dimensional gel electrophoresis. In the regenerating limbs without RA treatment, a few hundred kinds of proteins were found to be synthesized at the stage of wound healing and the total number of protein synthesized increased greatly as regeneration proceeded. The same trend was also observed in the RA-treated regenerating limbs. Interestingly, some protein spots were noted to be either newly synthesized or highly expressed by the RA treatment especially at the stage of dedifferentiation. The results shows that the enhancement of dedifferentiation state after the RA treatment correlates well with the protein synthesis profile, and suggest that those proteins are important for the RA-evoked pattern duplication in the regenerating limbs of salamander.

• 한국동물학회:뉴스레터
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• 제12권2호
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• pp.100.2-100
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• 1995

No Abstract, See Full Text

• 한국동물학회:학술대회논문집
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• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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• pp.228.2-228
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• 1995

No Abstract, See Full Text

• BMB Reports
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• 제54권7호
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• pp.356-367
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• 2021

Cell-based therapy is a promising approach in the field of regenerative medicine. As cells are formed into spheroids, their survival, functions, and engraftment in the transplanted site are significantly improved compared to single cell transplantation. To improve the therapeutic effect of cell spheroids even further, various biomaterials (e.g., nano- or microparticles, fibers, and hydrogels) have been developed for spheroid engineering. These biomaterials not only can control the overall spheroid formation (e.g., size, shape, aggregation speed, and degree of compaction), but also can regulate cell-to-cell and cell-to-matrix interactions in spheroids. Therefore, cell spheroids in synergy with biomaterials have recently emerged for cell-based regenerative therapy. Biomaterials-assisted spheroid engineering has been extensively studied for regeneration of bone or/and cartilage defects, critical limb ischemia, and myocardial infarction. Furthermore, it has been expanded to pancreas islets and hair follicle transplantation. This paper comprehensively reviews biomaterials-assisted spheroid engineering for regenerative therapy.

• Animal cells and systems
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• 제6권4호
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• pp.301-304
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• 2002

In our previous study, we have shown that Fgf-8 is expressed in the basal layer of the apical epithelial cap (AEC) and in the underlying thin layer of mesenchymal tissue of the regenerating limbs of Mexican axolotl, Amby-stoma mexicanum. Our present RT-PCR data also demonstrate that Fgf-8 transcript is localized both in the mesenchymal and epidermal tissues. To understand the effect of retinoic acid (RA) on the expression of Fgf-8 in the regenerating axolotl limbs, RA was injected intraperitoneally at the dediffer-entiation stage of limb regeneration. The RA treatment caused 8 change in the Fgf-8 expression profile of the regenerating limbs. In RA-treated limbs, duration of Fgf-8 expression was prolonged and a high level of expression was maintained during dedifferentiation and blastema formation stages. These results suggest that Fgf-8 is an important molecule in the process of pattern duplication of regenerating salamander limbs evoked by RA treatment.

• 한국발생생물학회지:발생과생식
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• 제2권1호
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• pp.53-62
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• 1998

리소솜 acid phosphatase(LAP)를 포함하는 리소솜 산성 가수 분해 효소는 세포 내,외 성분의 분해에 중요한 기능을 수행하며, 유미 양서류의 다리 재생 과정에서 LAP의 효소 활성도는 탈분화시기에 특이적으로 증가하는 것으로 알려져 있다. 본 연구에서는 멕시코산 도룡뇽의 일종인 axolotl(Ambystoma mexicanum)의 다리 재생 가정에서 정상적인 다리 재생 조직과 retinoic acid (RA)가 처리된 다리 재생 조직에서 LAP의 분포 및 발현을 면역 화학적 방법으로 조사하였다. Axolotl의 다리 재생 조직에서 한국산 도룡뇽(Hynobius leechii)의 LAP에 대한 단일 항체를 이용한 면역 반응도는 줄 탈분화시기에 있는 wound epidermis와 미분화된 중배엽성 세포인 blastema 세포, 탈분화 상태의 근육과 연골 부위 세포에서 나타나는 것으로 조사되었다. 또한 LAP의 면역 반응도는 재생단게의 초기에 점차 증가하여 탈분화시기 동안 최대치에 도달하였으며, 재분화가 시작되면서 basal level 까지 서서히 감소하는 것으로 나타나\ulcorner. 유미 양서류의 다리 재생 과정에서 골격 패턴의 복제를 유발시키는 것으로 알려져 있는 RA 처리는 LAP의 면역 반응도를 강화시키는 것으로 나타나\ulcorner. 즉, RA가 처리된 다리 재생 조직에서 LAP의 면역 반응도는 정상적인 다리 재생 조직에서 보다 강하게 나타났으며, 면역 반응도가 나타나는 시기 또한 연장되는 것으로 조사되었다. 이와 같은 결과는 다리 재생 조직에서 RA 처리가 탈분화시기의 연장에 관여한다는 것을 시사하고 있다.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1998년도 한국생물과학협회 학술발표대회
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• pp.234.1-234
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• 1998

No Abstract, See Full Text