RESUMO

The embryological origin of the trapezius and sternocleidomastoid muscles has been debated for over a century. To shed light on this issue, the present anatomical study was performed. Five fresh frozen human cadavers, three males and two females, were used for this study. Samples from each specimen's trapezius and sternocleidomastoid were fixed in 10% formalin and placed in paraffin blocks. As Paired like homeodomain 2 (Pitx2) and T-box factor 1(Tbx1) have been implicated in the region and muscle type regulation, we performed Tbx1 and Pitx2 Immunohistochemistry (IHC) on these muscle tissue samples to identify the origin of the trapezius and sternocleidomastoid muscles. We have used the latest version of QuPath, v0.4.3, software to quantify the Tbx and Pitx2 staining. For the sternocleidomastoid muscle, for evaluated samples, the average amount of positively stained Tbx1 and Pitx2 was 25% (range 16%-30%) and 18% (range 12%-23%), respectively. For the trapezius muscles, for evaluated samples, the average amount of positively stained Tbx1 and Pitx2 parts of the samples was 17% (range 15%-20%) and 15% (14%-17%), respectively. Our anatomical findings suggest dual origins of both the trapezius and sternocleidomastoid muscles. Additionally, as neither Pitx2 nor Tbx1 made up all the staining observed for each muscle, other contributions to these structures are likely. Future studies with larger samples are now necessary to confirm these findings.

Assuntos

RESUMO

qRT-PCR requires reliable internal control genes stably expressed in different samples and experimental conditions. The stability of reference genes is rarely tested experimentally, especially in developing tissues given the singularity of these samples. Here we evaluated the suitability of a set of reference genes (Actb, Gapdh, Tbp, Pgk1 and Sdha) using samples from early mouse embryo tissues that are widely used in research (somites, prosencephalon and heart) at different developmental stages. The comparative ΔCq method and five software packages (NormFinder, geNorm, BestKeeper, DataAssist and RefFinder) were used to rank the most stable genes while GenEx and GeNorm programs determined the optimal total number of reference genes for a reliable normalization. The ranking of most reliable reference genes was different for each tissue evaluated: (1) in somite from embryos with 16-18 somite pairs stage, the combination of Pgk1 and Actb provided the best normalization and Actb also presented high stability levels at an earlier developmental stage; (2) Gapdh is the most stable gene in prosencephalon in the two developmental stages tested; and (3) in heart samples, Sdha, Gapdh and Actb were the best combination for qPCR normalization. The analysis of these three tissues simultaneously indicated the combination of Gapdh, Actb and Tbp as the most reliable internal control. This study highlights the importance of appropriate reference genes according to the cell type and/or tissue of interest. The data here described can be applied in future research using mouse embryos as a model for mammalian development.

Assuntos

RESUMO

BACKGROUND: In most vertebrates, the segmentation of the paraxial mesoderm involves the formation of metameric units called somites through a mesenchymal-epithelial transition. However, this process is different in Xenopus laevis because it does not form an epithelial somite. Xenopus somitogenesis is characterized by a complex cells rearrangement that requires the coordinated regulation of cell shape, adhesion, and motility. The molecular mechanisms that control these cell behaviors underlying somite formation are little known. Although the Paraxis has been implicated in the epithelialization of somite in chick and mouse, its role in Xenopus somite morphogenesis has not been determined. RESULTS: Using a morpholino and hormone-inducible construction approaches, we showed that both gain and loss of function of paraxis affect somite elongation, rotation and alignment, causing a severe disorganization of somitic tissue. We further found that depletion or overexpression of paraxis in the somite led to the downregulation or upregulation, respectively, of cell adhesion expression markers. Finally, we demonstrated that paraxis is necessary for the proper expression of myotomal and sclerotomal differentiation markers. CONCLUSIONS: Our results demonstrate that paraxis regulates the cell rearrangements that take place during the somitogenesis of Xenopus by regulating cell adhesion. Furthermore, paraxis is also required for somite differentiation.

Assuntos

RESUMO

Two paternal broiler lines (LL and LLc, originated from the same stock in 1998) of Embrapa Suinos e Aves genetic program were used. Line LL had been selected for eight generations for rapid growth while LLc had been mated randomly, without selection, as a control line. In the last generation, 30 males and 240 females were used in hierarchical matings with pedigree control to determine sire effect. The same number of birds were used, but matings were at random for LLc. Eggs of both lines were incubated and embryos were collected in different development stages (40, 45, 50, 55 and 60 hours after setting) in order to determine somite number (NOS). The number of evaluated embryos were 1,967 and 498 for LL and LLc, respectively. This study aimed to determine the changes in somite formation during embryonic development that resulted from artificial selection for heavier body weight. In the evaluated generation, the differences for body weight, feed conversion ratio, carcass and breast yields between LL and LLc were 546 g; 0.16 FC units, 2.20 and 2.19%, respectively. NOS was greater in the initial (40 h) and in the final (60 h) stages for LL and in the intermediary stages (45, 50 and 55 h) for LLc, indicating significant differences in the mechanisms of somite formation and development. Considering that both lines had the same genetic origin, such deviation indicates genetic changes due to artificial selection for body weight. The heritability estimates indicated low additive genetic effect for NOS in the different stages. Since the alterations were due to artificial selection for body weight, one could consider the hypothesis of using such genetic marker throughout family selection.

RESUMO

Two paternal broiler lines (LL and LLc, originated from the same stock in 1998) of Embrapa Suinos e Aves genetic program were used. Line LL had been selected for eight generations for rapid growth while LLc had been mated randomly, without selection, as a control line. In the last generation, 30 males and 240 females were used in hierarchical matings with pedigree control to determine sire effect. The same number of birds were used, but matings were at random for LLc. Eggs of both lines were incubated and embryos were collected in different development stages (40, 45, 50, 55 and 60 hours after setting) in order to determine somite number (NOS). The number of evaluated embryos were 1,967 and 498 for LL and LLc, respectively. This study aimed to determine the changes in somite formation during embryonic development that resulted from artificial selection for heavier body weight. In the evaluated generation, the differences for body weight, feed conversion ratio, carcass and breast yields between LL and LLc were 546 g; 0.16 FC units, 2.20 and 2.19%, respectively. NOS was greater in the initial (40 h) and in the final (60 h) stages for LL and in the intermediary stages (45, 50 and 55 h) for LLc, indicating significant differences in the mechanisms of somite formation and development. Considering that both lines had the same genetic origin, such deviation indicates genetic changes due to artificial selection for body weight. The heritability estimates indicated low additive genetic effect for NOS in the different stages. Since the alterations were due to artificial selection for body weight, one could consider the hypothesis of using such genetic marker throughout family selection.