학술논문
SMJEŠTAJ TFF1 I TFF3 PROTEINA U RAZLIČITIM STADIJIMA RAZVOJA MIŠJEG ZAMETKALocalization of TFF1 and TFF3 proteins at different stages of the mouse embryonic development
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TEXT
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Source
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Language
Croatian
Abstract
Cilj istraživanja: TFF proteini uključeni su u razne fiziološke procese u probavnom, dišnom, mokraćnom i živčanom sustavu. Osim toga, prisutni su i u patološkim stanjima. Utječu na migraciju, diferencijaciju i apoptozu stanica, kao i na angiogenezu i imunološki odgovor te se intenzivno istražuju u zdravlju i bolesti. S obzirom da ima vrlo malo podataka o ekspresiji TFF proteina tijekom embrionalnog razvoja, u ovom radu imunohistokemijskom metodom istražena je prisutnost TFF1 i TFF3 proteina u tkivima mišjih zametaka starosti od 14 do 18 dana. Analizirani su i histomorfološki parametri epifizne ploče rasta i sekundarnih centara okoštavanja na proksimalnom dijelu tibije divljeg tipa i miševa s isključenim genom TFF3 starih mjesec dana. Materijal i metode: Imunohistokemijski pokusi izvedeni su na preparatima 23 zametka CD1 miševa fiksiranih 4%-nim paraformaldehidom i uklopljenih u parafin, starosti od 14 do 18 dana. Histomorfometrijski parametri analizirani su na tibijama miševa divljeg tipa visoko srođenih životinja razvijenih križanjem sojeva 129/Sv i C57BL/6j te miševa s isključenim genom TFF3. Korišteno je 5 miševa po skupini, a histološki preparati tibija obojeni su trikromnim bojenjem po Massonu. Rezultati: Prisutnost TFF 1 i 3 proteina u mišjim zamecima uočena je u sluznici probavnog i dišnog sustava, gušterači, kanalićima bubrega, epidermisu kože, živčanom sustavu te okoštavanju, a pojavljivala se ovisno o stadiju razvoja miša i stupnju diferencijacije tkiva. Smještaj TFF1 i TFF3 proteina u tkivima mišjih zametaka nije bio povezan s podrijetlom tkiva prema embrionalnim zametnim listićima. Nisu pronađene značajne histomorfološke razlike u debljini epifizne ploče rasta, pojedinih zona enhondralnog okoštavanja niti u gustoći hondrocita po jedinici površine između divljeg tipa miševa i miševa s isključenim genom TFF3, međutim, u sekundarnim centrima okoštavanja u epifizama mišjih tibija u u životinja s isključenim genom TFF3 vrijednosti gustoće volumena, gustoće površine i broja koštanih gredica bile su znatno manje, a razdvojenost koštanih gredica znatno veća. Zaključak: TFF1 i TFF3 protein zbog svojih brojnih fizioloških uloga važni su u razvoju zametka te u nekim slučajevima mogu poslužiti kao markeri fiziološkog sazrijevanja tkiva. TFF3 vjerojatno ima važan učinak na stvaranje i kvalitetu spužvaste kosti u sekundarnim centrima okoštavanja.
Objectives: TFF proteins are involved in different physiological processes in the digestive, respiratory, urinary and nervous system and are also present in different pathological conditions. These proteins influence cell migration, differentiation and apoptosis, as well as angiogenesis and immune response and are under active research in health and disease. Since very limited data are available on the expression of TFF proteins in the embryonic tissues, presence of TFF1 and TFF3 in the tissues of 14 to 18-day old mice was investigated in this work. Also, histomorphological parameters of the epiphyseal growth plate and of the secondary ossification centers were investigated in the proximal tibia of the wild type and TFF3 knock-out mice. Material and methods: Immunohistochemical experiments were done manually, using 23 formalin fixed, paraffin embedded CD1 mouse embryos, 14 to 18-day old. Histomorphometric parameters were analyzed in the tibiae of inbred wild-type mice developed by crossing 129/Sv and C57BL/6j strain and TFF3 knock-out mice. 5 mice per group were used, and histological sections of tibiae were stained using Masson’s trichrome stain. Results: Presence of TFF1 and TFF3 proteins was detected in the gastrointestinal and respiratory mucosa, pancreas, kidney tubules, epidermis, nervous system and areas of ossification of mouse embryos and occurred depending on the stage of embryonic development and tissue differentiation. Localization of TFF1 and TFF3 protein in mouse embryonic tissues didn’t depend on the germinal layer from which the tissue originated. No significant histomorphological differences between wild-type mice and TFF3 knock out mice were found in epiphyseal plate thickness, in the thickness of different zones of endochondral ossification, and in the chondrocyte density. However, cancellous bone in the secondary ossification centers of mouse tibia epiphyses showed significantly lower values of volume density, surface density and trabecular number, while trabecular separation was significantly higher. Conclusion: Because of their numerous physiological roles, TFF1 and TFF3 are important in the embryonic development, and in some cases may serve as early markers of tissue maturation. TFF3 probably has an important effect on the formation and quality of the cancellous bone in the secondary ossification centers.
Objectives: TFF proteins are involved in different physiological processes in the digestive, respiratory, urinary and nervous system and are also present in different pathological conditions. These proteins influence cell migration, differentiation and apoptosis, as well as angiogenesis and immune response and are under active research in health and disease. Since very limited data are available on the expression of TFF proteins in the embryonic tissues, presence of TFF1 and TFF3 in the tissues of 14 to 18-day old mice was investigated in this work. Also, histomorphological parameters of the epiphyseal growth plate and of the secondary ossification centers were investigated in the proximal tibia of the wild type and TFF3 knock-out mice. Material and methods: Immunohistochemical experiments were done manually, using 23 formalin fixed, paraffin embedded CD1 mouse embryos, 14 to 18-day old. Histomorphometric parameters were analyzed in the tibiae of inbred wild-type mice developed by crossing 129/Sv and C57BL/6j strain and TFF3 knock-out mice. 5 mice per group were used, and histological sections of tibiae were stained using Masson’s trichrome stain. Results: Presence of TFF1 and TFF3 proteins was detected in the gastrointestinal and respiratory mucosa, pancreas, kidney tubules, epidermis, nervous system and areas of ossification of mouse embryos and occurred depending on the stage of embryonic development and tissue differentiation. Localization of TFF1 and TFF3 protein in mouse embryonic tissues didn’t depend on the germinal layer from which the tissue originated. No significant histomorphological differences between wild-type mice and TFF3 knock out mice were found in epiphyseal plate thickness, in the thickness of different zones of endochondral ossification, and in the chondrocyte density. However, cancellous bone in the secondary ossification centers of mouse tibia epiphyses showed significantly lower values of volume density, surface density and trabecular number, while trabecular separation was significantly higher. Conclusion: Because of their numerous physiological roles, TFF1 and TFF3 are important in the embryonic development, and in some cases may serve as early markers of tissue maturation. TFF3 probably has an important effect on the formation and quality of the cancellous bone in the secondary ossification centers.