| In 1979, Brown GH and Wolken JJ systematically summarized entire research proceedings on liquid crystalline in biological organism in their book "Liquid Crystals and Biological Structures". The book became an authoritative reference in the research field. In 1978 and 1979, findings during chicken development have caught researchers'attention. For the first time, the work revealed that liquid crystalline exists massively in liver, yolk sac, blood, and many other more developing tissues and organs during chicken embryogenesis. After these the field seems very quiet for more than two decades. Rationale caused this silence is not because importance of liquid crystal had been doubted. Reason is how to link what we knew on liquid crystal to current biological and biomedical knowledge. The following questions could be considered. Among vertebrate physiological processes, how liquid crystal functions in cells, tissues or organs? Is any disease related with Liquid crystal disorder/defect; can appearance of it be developed as an early biomarker to represent or predict the early syndrome of human disease? In life origin or embryonic development, does liquid crystal play its function with the same molecular mechanism? Major question on this issue could be focus on what is the comprehension of molecular biology. Is any genes related with appearance or disappearance during normal or abnormal biological processes? Are these same genes related to any human diseases? What are the best models or are there any best models for us to explore these questions?In the recent years, some research reports raised public attention on topics of life origin, organ morphogenesis, and pathogenesis of human cardiac disease, eye disease, renal disease and cerebrovascular dysfunction. In 2007, Nakata M et al revealed for the first time that small base pair double strand oligos (5-20bp) can form nematic arid columnar liquid crystals in water. This finding provides a key linker in the emergence of life on earth. Helary C et al in 2005, Giraud-Guille MM et al in 2008 an d others revealed that, in liquid crystal collagen 3D culture system, ECM protein including, stem cell from bone differentiated to an epithelium-like cells promoted by extracellular matrices. Meanwhile, application of liquid crystal such as liquid-crystal monitoring stem cell differentiation became one of scientific inventions that caught the most attention in community in 2006. To contribute our research results to this field, our studies presented here are mainly categorized as four parts. The works were carried out with two animal models, avian development and mouse transgenic model.(?) In vivo analysis in chicken embryogenesis and in vitro mimicking of hepatic LCLD. The data broaden our understanding for human Steatohepatitis or fatty-liver disease. Comprehension could direct us to develop a new diagnostics procedure. Using X-ray diffraction, small angle X-ray scattering and phase transition array, we concluded that:(A) the liquid crystallization in liver is routine biological process during avian embryogenesis. Our inspection covers chicken, duck, pigeon and Taihe fowl. The liquid crystallization occurred at earlier stage and the massive hepatic LCLD accumulated in hepatocytes till later stage. In first two weeks of avian postnatal development the hepatic LCLD would be absorbed gradually. (B) Small angle X-ray scattering revealed that the periodic spaces of liquid crystalline are distributed from 36 to 39 A. In all avian embryonic livers we inspected, the small angle scattering peak can be detected directly from fresh embryonic livers. (C) X-ray diffraction combining thermal measurement revealed that the phase transitions between liquid crystalline, crystalline and anisotropic states can take place in the avian hepatic LCLD. (D) Based on component analysis of chicken hepatic LCLD, we constructed the liquid crystal lipid droplet in vitro. The mimicked hepatic LCLDs are in possession of all characteristics that we described above.(?) Characteristics of renal LCLD in chicken embryogenesis. LCLD analysis during renal development from mesonephros to metanephros indicated that LCLD is very similar to the birefringence particles, which has become a typical diagnostic characteristic used to evaluate the kidney complications of Anderson-Fabry Disease. The disease is an X-linked recessive disorder located on chromosome Xq22.1 characterized by a deficiency of the lysosomal enzyme?-galactosidase A. Through the use of polarization microscopy and electron microscopy, we demonstrated (A) The Maltese crosses are initially observed in E8 in mesonephros. The number of these Maltese crosses increases during development of the embryo and the size of the birefringent myelin bodies continuously enlarge until E13; (B) in the metanephros phase, the Maltese crosses are first observed in E14 embryos. Both the number and size of the Maltese crosses increase with the development of the embryo as well; (C) our ultrastructural studies demonstrated that the Maltese cross is a typical configuration of myelin bodies with concentric lamellation situated in the cytoplasm of proximal tubular epithelial cells and the lumen of proximal tubules in both mesonephros and metanephros tissues.(?) LCLD and Calcium Preservation in Yolk Sac development. In this part of our study, we demonstrated that abundant liquid crystal droplets (LCDLs) subsist with calcium carbonate vaterite in the yolk sac during chicken development and 'revealed a potential function of LCDL in calcium carbonate crystallization. We hypothesized that the accumulation of calcium carbonate on lipid lamina in LCDLs is the major mechanism for calcium preservation in yolk sac during avian embryogenesis. Our finding facilitates the comprehension of the function of LCDLs in animal development, an advancement combined with the analysis on the co-subsistence of LCDLs and calcium carbonate calcite, aragonite and vaterite crystal in several organisms that offer valuable understanding for human calcium stone disease. It may also benefit to make highly efficient calcium carbonate involved microcapsule for drug delivery system.(?) Alb-p37 transgenic mouse model reveals liquid crystal involved Steatohepatitis disease linked to cardiac hypotrophy and arrhythmia triggered by high protein diet. To investigate function of liquid crystalline in animal development we identified three hepatic liquid-crystalline related proteins p37, p25 and p18 from newborn and E18 embryonic chicken. Accordingly we generated liver specific p37 overexpression transgenic mouse model. Although the Alb-p37 mice appear normal on histology and behave analysis, their abnormality was revealed when they were on different diet. Under the high protein diet, the Alb-p37 mice exhibit the following disorders on their liver size, body weight and heart size. These results indicated that p37 protein could be a critical factor linking steatohepatitis and cardiac hypotrophy with the involvement of hepatic liquid crystalline.Utilizing Alb-p37 transgenic animal model, we concluded the following results. (A) On normal diet condition, the mice have no any observable morphology phenotype. However on high protein diet, the liver-specific expression of p37 driven by Alb promoter directly results in liver enlargement with massive hepatic liquid crystalline in hepatocytes. (B) Although no any liquid crystalline could be observed in heart and other tissues or organs of Alb-p37 transgenic animals, significant enlargement were observed on their heart and body-sizes. (C) With the increase of heart and body sizes, the disorder of their cardiac function was observed. Compared to their littermates, the ECGs revealed cardiac dysfunction or heart beat abnormality in the transgenic mice. The heart beat slowing-down w as characterized as severer arrhythmia bradicardia. (D) Under the treadmill-exercise condition, the arrhythmia bradicardia developed to premature QRS complex that leads to sudden cardiac death. |
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