Chamber specific expression of Myosin heavy chain 7b in the heart of vertebrates

In extant vertebrates, myosin heavy chain (MyHC) 6 and 7 are the main isoforms of atrial and ventricular myocardium respectively, whereas MyHC7b has been proposed to be an ancient cardiac isoform only expressed during embryonic development in modern species. In preliminary immunohistochemical studies of the heart of the lesser spotted dogfish (Scyliorhinus canicula; Chondrichthyes), we have observed that while MF20 labels homogeneously all the myocardium, A4.1025 labels the inflow cardiac segments (sinus venosus and atrium) but not the outflow segments (ventricle and conus arteriosus). In order to interpret these results, we have performed western and slot blots from samples of dogfish and hamster (Mesocricetus auratus) hearts, HPLC-ESI-MS/MS from dogfish samples, and immunohistochemistry in hearts of representative species of vertebrates, namely elasmobranchs, polypteriforms, acipenseriforms, teleosts and mammals, using MF20 and A4.1025 antibodies. Western and slot blot results confirmed the specificity of MF20 and A4.1025 for MyHC in dogfish, as well as their differential reactivity against different myocardial segments. HPLC-ESI-MS/MS using protein databases from Callorhinchus milii (Chondrichthyes) and Chordata revealed the presence of MyHC6 and 7 in all the dogfish myocardial segments, and of MyHC7b only in the outflow segments. Immunohistochemistry showed that while MF20 signals were homogeneous in all the myocardial segments of all the species studied, A4.1025 signals were restricted to the inflow myocardial segments in elasmobranchs, homogeneous in teleosts and acipenseriforms, and low in the ventricle of polypteriforms. It can be inferred that the A4.1025 antibody, as opposed to MF20, has a low affinity for MyHC7b, at least in the dogfish. In addition, we show that MyHC distribution in the cardiac chambers has changed during the evolution of gnathostomes.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech.Study supported by grants CGL2017-85090-P and CGL2014-52356-P (Ministerio de Economía y Competitividad), FPU15/03209 (Ministerio de Educación, Cultura y Deporte), contract UMAJI75 (Junta de Andalucía, European Social Found), FEDER and Universidad de Málaga

Myocardial trabeculation in embryos of Scyliorhinus canicula (Elasmobranchii, Chondrichthyans)

Currently, three types of ventricular myoarchitecture are recognized in vertebrates, namely compact, spongy (trabeculated) and mixed myocardium. Mixed myocardium, which has been recently proposed as the primitive condition in gnathostomes, is composed of two myocardial layers: an inner trabeculated and an outer compact one. The trabeculation process has been studied in teleosts, showing exclusively spongy myocardium, and mammals and birds, characterized by a compact myocardial ventricular wall. In zebrafish, mouse and chicken embryos, the trabeculae develop as luminal myocardial ridges protruding into the lumen. In mammals and birds, further compactation of trabeculae leads to the formation of a compact layer. The potential mechanisms that may contribute to the formation of the ridges are under discussion and include myocardial proliferation, endocardial invagination, and bending of the entire myocardial layer. However, no description of the development of the mixed myocardium is available. To shed some light on this issue, we have studied the heart development of an elasmobranch species with mixed myocardium, the lesser spotted dogfish (Scyliorhinus canicula; Chondrichthyes), by means of histological and immunohistochemical techniques for light microscopy, semithin sections, scanning electron microscopy and transmission electron microscopy. Our results suggest that in the dogfish the intertrabecular spaces develop by connections between early intramyocardial spaces and the lumen of the ventricle through invaginations of the endocardial line. Chondrichthyans are the earliest diverged lineage of gnathostomes and, consequently, they have the most primitive cardiac design. Although chicken, mouse, and recently zebrafish have been considered powerful vertebrate models to study heart development, we propose that the trabeculation process in the dogfish is representative of the early steps of the ventricular morphogenesis in vertebrates.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech.Study supported by grant CGL2017-85090-P and CGL2014-52356-P (Ministerio de Economía y Competitividad), FPU15/03209 (Ministerio de Educación, Cultura y Deporte), FEDER, and Universidad de Málaga

Emotion Recognition in Horses with Convolutional Neural Networks

Creating intelligent systems capable of recognizing emotions is a difficult task, especially when looking at emotions in animals. This paper describes the process of designing a "proof of concept" system to recognize emotions in horses. This system is formed by two elements, a detector and a model. The detector is a fast region-based convolutional neural network that detects horses in an image. The model is a convolutional neural network that predicts the emotions of those horses. These two elements were trained with multiple images of horses until they achieved high accuracy in their tasks. In total, 400 images of horses were collected and labeled to train both the detector and the model while 40 were used to test the system. Once the two components were validated, they were combined into a testable system that would detect equine emotions based on established behavioral ethograms indicating emotional affect through head, neck, ear, muzzle and eye position. The system showed an accuracy of 80% on the validation set and 65% on the test set, demonstrating that it is possible to predict emotions in animals using autonomous intelligent systems. Such a system has multiple applications including further studies in the growing field of animal emotions as well as in the veterinary field to determine the physical welfare of horses or other livestock.Comment: 14 pages, 11figure

Hamsters’ (mesocricetus auratus) memory in a radial maze analog: the role of spatial versus olfactory cues

The golden hamster’s (Mesocricetus auratus) performance on radial maze tasks has not been studied a lot. Here we report the results of a spatial memory task that involved eight food stations equidistant from the center of a circular platform. Each of six male hamsters depleted the food stations along successive choices. After each choice and a 5-s retention delay, the hamster was brought back to the center of the platform for the next choice opportunity. When only one baited station was left, the platform was rotated to evaluate whether olfactory traces guided hamsters’ choices. Results showed that despite the retention delay hamsters performed above chance in searching for food. The choice distributions observed during the rotation probes were consistent with spatial memory and could be explained without assuming guidance by olfactory cues. The radial maze analog we devised could be useful in furthering the study of spatial memory in hamsters.Francois Tonneau was supported by the Portuguese FCT during the writing of this article

Ventricular myocardial trabeculation in chondrichthyans. Evolutionary implications

The formation of trabeculae (trabeculation) in vertebrates occurs during cardiac development in the three structural types of ventricular myocardium, namely, compact, spongy and mixed. The compact type is mainly composed of compacted muscular fibers and the spongy type of muscular trabeculae. The mixed type, with an inner trabecular and an outer compact layer, has been proposed as the primitive condition in gnathostomes. In vertebrate models, trabeculation initiates following two alternative mechanisms: (1) in chicken and mouse, the endocardial cells evaginate towards the two-layered myocardium; (2) in zebrafish, cardiomyocytes from the mono-layered myocardium invaginate towards the endocardium. Trabeculation in the mixed myocardium has not been described yet. We have studied the mixed myocardium formation in the dogfish (Scyliorhinus canicula, Elasmobranchii) using light, scanning and transmission electron microscopy. At stage 27, the ventricle consists of a two-layered myocardium internally lined by endocardium, both separated by cardiac jelly. Trabeculation starts at stage 28, when small spaces between cardiomyocytes appear, the cardiac jelly become thinner and the endocardium focally contacts the myocardium. At stage 29 the spaces between cardiomyocytes increase in size and get lined by the endocardium, shaping the presumptive trabeculae. At later stages, the trabeculae increase in complexity and the outer cardiomyocytes proliferate and get compacted, delineating the definitive trabeculated and compact myocardia. We conclude that early trabeculation in elasmobranchs matches that described in tetrapods. Thus, the mechanism of trabeculation of the mixed ventricular myocardium has been conserved in the formation of the compact myocardium of tetrapods. Additional studies in actinopterygians with different myocardial types may uncover how trabeculation has evolved during the evolution of gnathostomes.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. CGL2017-85090-P, FPU15/03209, UMAJI75 and FEDER