Antagonistic regulation of homeologous uncx.L and uncx.S genes orchestrates myotome and sclerotome differentiation in the evolutionarily divergent vertebral column of Xenopus laevis.

In anurans, the vertebral column diverges widely from that of other tetrapods; yet the molecular mechanisms underlying its morphogenesis remain largely unexplored. In this study, we investigate the role of the homeologous uncx.L and uncx.S genes in the vertebral column morphogenesis of the allotetraploid frog Xenopus laevis. We initiated our study by cloning the uncx orthologous genes in the anuran Xenopus and determining their spatial expression patterns using in situ hybridization. Additionally, we employed gain-of-function and loss-of-function approaches through dexamethasone-inducible uncx constructs and antisense morpholino oligonucleotides, respectively. Comparative analysis of the messenger RNA sequences of homeologous uncx genes revealed that the uncx.L variant lacks the eh1-like repressor domain. Our spatial expression analysis indicated that in the presomitic mesoderm and somites, the transcripts of uncx.L and uncx.S are located in overlapping domains. Alterations in the function of uncx genes significantly impact the development and differentiation of the sclerotome and myotome, resulting in axial skeleton malformations. Our findings suggest a scenario where the homeologous genes uncx.L and uncx.S exhibit antagonistic functions during somitogenesis. Specifically, uncx.S appears to be crucial for sclerotome development and differentiation, while uncx.L primarily influences myotome development. Postallotetraploidization, the uncx.L gene in X. laevis evolved to lose its eh1-like repressor domain, transforming into a "native dominant negative" variant that potentially competes with uncx.S for the same target genes. Finally, the histological analysis revealed that uncx.S expression is necessary for the correct formation of pedicles and neural arch of the vertebrae, and uncx.L is required for trunk muscle development.

Ontogeny of the meniscus in the anuran Xenopus laevis.

The anuran knee joint is subjected to the jump, one of the tetrapods' most demanding mechanical stresses. Consistent with this continuous effort, the knee of the anurans has a complex structure comparable to that of an amniote. Here, we describe the ontogeny of the Xenopus knee tissues and study the morphogenesis of the knee joint shape by performing a geometric morphometric analysis of specially selected anatomical structures: the menisci and the long bone epiphyses. A meniscus is a crescent-shaped fibrocartilaginous structure, with a triangular cross-section inserted between joints surfaces. A meniscus transmits load across the tibiofemoral joint by increasing congruity of the long bone epiphysis and decreasing the resulting stress exerted on the articular cartilage. We ask two questions: (1) what is the tissue composition along the ontogeny of the menisci of a swimming frog? (2) How do the menisci acquire the shape that will allow their adjustment? We studied the structures and tissue ontogeny of the knee of several specimens of Xenopus laevis and evaluated the congruity of the knee structures across the species ontogeny. Histological sections showed that the cavitation process responsible for separating the menisci and the epiphyses seems to be pivotal in shaping the conformity of these structures and the long bone epiphyses of the hindlimbs. The geometric morphometric analysis allowed us to interpret three phases of differentiation associated with limb functionality. The characteristic shape of the meniscus appears early in the ontogeny of the knee, simultaneously with the epiphysis contours.

A novel endothelium-independent effect of insulin on basal vascular tone in cafeteria diet-induced hypertensive rats.

Insulin vasorelaxant effect in metabolic syndrome has been shown on precontracted vessels. However, the insulin effects on basal vascular tone and its interrelationship with nitric oxide (NO) and K-channels are unknown. To test the effect of insulin on the basal vascular tone in isolated aortic rings from the cafeteria diet-induced hypertensive rats and to determine the role of NO and K-channels on this insulin effect. Male Wistar rats were randomized into two groups: one group fed with a cafeteria diet (CafR) and another fed with a standard chow diet (control rats: CR). Then, in isolated aortic rings, the insulin effect on the basal tone and the role of K-channels were evaluated. Also, the endothelial function, NO levels, and resting membrane potential were measured. CafR increased blood pressure (138 ± 6.2 mmHg; n = 9 vs. CR: 109 ± 1.4 mmHg; n = 9; p < 0.001) and vascular basal tone. Insulin 400 mU/ml reduced basal tone in aortic rings (-284 ± 47 mg; n = 9). This effect was unaffected by endothelium removal or NG-nitro-l-arginine methyl ester (L-NAME) treatment. Likewise, CafR showed low NO levels and a hyperpolarized resting membrane potential. Insulin decreased the resting membrane potential and the KCa and Kv channels blockers abolished this effect. In CafR, endothelial dysfunction is accompanied by an increased basal tone. Insulin reduced it by Kv and KCa channels dependent mechanisms, using an endothelium-independent pathway. These results highlight a novel insulin effect on basal tone of aortic rings from animals with metabolic syndrome and endothelial dysfunction, pathophysiological conditions associated with human hypertension.

Delineating the anuran axial skeleton.

The axial skeleton of the anurans has undergone an evolutionary reduction of its bone elements. This structural plan is strongly preserved throughout the order and would have emerged as a highly specialized anatomical adaptation to its locomotor jumping pattern. The development programs that direct the vertebral morphogenesis of the anurans are poorly described and the molecular bases that have caused their pattern to differ from other tetrapods are completely unknown. In this work, we review the ontogeny of the spinal column of the anurans and explore the genetic mechanisms that could explain the morphological difference and the maintenance of the body plan during evolution. Here, we propose that the absence of caudal osseous elements, as a consequence of the inability of sclerotomes to form cartilaginous condensations in frogs, could be due to changes in both pattern and expression levels of Hox, Pax1, Pax9 and Uncx4.1 genes along the anteroposterior axis. The anteriorised expression of the Hox genes together with the reduction in the expression levels of Pax1, Pax9 and Uncx4 in the posterior somites could explain, at least partly, the loss of caudal vertebrae in the anurans during evolution.