1. Investigación

Flipped classroom (FC) is a teaching method where traditional learning roles are inverted. Students are provided with material in advance and are expected to study the content prior to in-class sessions. These sessions are subsequently utilized to clarify doubts and examine in greater depth the previously acquired knowledge. Despite the widespread nature of its approach in health education, its application in basic veterinary subjects remains poorly described. This study explores the implementation of the FC approach in veterinary physiology, biochemistry, anatomy, and embryology. Pre-class material was mainly provided in video format, and class sessions facilitated quizzes and interactive activities aimed to reinforce understanding. The findings indicate a high level of student involvement and effective class preparation, as evidenced by over 84% of students participating in FC in-class sessions and generally achieving satisfactory scores on quizzes. A survey conducted at the end of the first semester shows that a high proportion of students positively valued pre-class material (>90%), quizzes (82%), and the FC approach (66%). However, by the end of the second semester, traditional lectures were preferred by more students than FC (45% and 25%, respectively), while 30% of the students mentioned having no preference between the two methods. Analysis of open-ended responses underscored positive facets of the FC approach, including self-organization, enhanced understanding, and availability of pre-class material. However, it also emphasized challenges associated with FC, such as the significant time and effort required. In conclusion, this study suggests that the FC approach can be well received in integrated basic veterinary subjects if it does not imply an excessive student workload, underscoring the potential benefits of a blended teaching approach that combines elements of both traditional and FC methods.

The forebrain includes the cerebral cortex, the thalamus, and the striatum and globus pallidus (GP) in the subpallium. The formation of these structures and their interconnections by specific axonal tracts take place in a precise and orchestrated time and spatial-dependent manner during development. However, the knowledge of the molecular and cellular mechanisms that are involved is rather limited. Moreover, while many extracellular cues and specific receptors have been shown to play a role in different aspects of nervous system development, including neuron migration and axon guidance, examples of intracellular signaling effectors involved in these processes are sparse. In the present work, we have shown that the atypical RhoGTPase, Rnd3, is expressed very early during brain development and keeps a dynamic expression in several brain regions including the cortex, the thalamus, and the subpallium. By using a gene-trap allele (Rnd3gt) and immunological techniques, we have shown that Rnd3gt/gt embryos display severe defects in striatal and thalamocortical axonal projections (SAs and TCAs, respectively) and defects in GP formation already at early stages. Surprisingly, the corridor, an important intermediate target for TCAs is still present in these mutants. Mechanistically, a conditional genetic deletion approach revealed that Rnd3 is primarily required for the normal development of Medial Ganglionic Eminence-derived structures, such as the GP, and therefore acts non-cell autonomously in SAs and TCAs. In conclusion, we have demonstrated the important role of Rnd3 as an early regulator of subpallium development in vivo and revealed new insights about SAs and TCAs development.

Neural stem cells directly or indirectly generate all neurons and macroglial cells and guide migrating neurons by using a palisade-like scaffold made of their radial fibers. Here, we describe an unexpected role for the radial fiber scaffold in directing corticospinal and other axons at the junction between the striatum and globus pallidus. The maintenance of this scaffold, and consequently axon pathfinding, is dependent on the expression of an atypical RHO-GTPase, RND3/RHOE, together with its binding partner ARHGAP35/P190A, a RHO GTPase-activating protein, in the radial glia-like neural stem cells within the ventricular zone of the medial ganglionic eminence. This role is independent of RND3 and ARHGAP35 expression in corticospinal neurons, where they regulate dendritic spine formation, axon elongation, and pontine midline crossing in a FEZF2-dependent manner. The prevalence of neural stem cell scaffolds and their expression of RND3 and ARHGAP35 suggests that these observations might be broadly relevant for axon guidance and neural circuit formation.

Glucose uptake into the mammalian nervous system is mediated by the family of facilitative glucose transporter proteins (GLUT). In this work we investigate how the expression of the main neuronal glucose transporters (GLUT3, GLUT4 and GLUT8) is modified during cerebellar cortex maturation. Our results reveal that the levels of the three transporters increase during the postnatal development of the cerebellum. GLUT3 localizes in the growing molecular layer and in the internal granule cell layer. However, the external granule cell layer, Purkinje cell cytoplasm and cytoplasm of the other cerebellar cells lack GLUT3 expression. GLUT4 and GLUT8 have partially overlapping patterns, which are detected in the cytoplasm and dendrites of Purkinje cells, and also in the internal granule cell layer where GLUT8 displays a more diffuse pattern. The differential localization of the transporters suggests that they play different roles in the cerebellum, although GLUT4 and GLUT8 could also perform some compensatory or redundant functions. In addition, the increase in the levels and the area expressing the three transporters suggests that these roles become more important as development advances. Interestingly, the external granule cells, which have been shown to express the monocarboxylate transporter MCT2, express none of the three main neuronal GLUTs. However, when these cells migrate inwardly to differentiate in the internal granule cells, they begin to produce GLUT3, GLUT4 and GLUT8, suggesting that the maturation of the cerebellar granule cells involves a switch in their metabolism in such a way that they start using glucose as they mature.

Peripheral nerve axotomy in adult mice elicits a complex response that includes increased glucose uptake in regenerating nerve cells. This work analyses the expression of the neuronal glucose transporters GLUT3, GLUT4 and GLUT8 in the facial nucleus of adult mice during the first days after facial nerve axotomy. Our results show that whereas GLUT3 levels do not vary, GLUT4 and GLUT8 immunoreactivity increases in the cell body of the injured motoneurons after the lesion. A sharp increase in GLUT4 immunoreactivity was detected 3 days after the nerve injury and levels remained high on Day 8, but to a lesser extent. GLUT8 also increased the levels but later than GLUT4, as they only rose on Day 8 post-lesion. These results indicate that glucose transport is activated in regenerating motoneurons and that GLUT4 plays a main role in this function. These results also suggest that metabolic defects involving impairment of glucose transporters may be principal components of the neurotoxic mechanisms leading to motoneuron death.

Rho proteins are a large family of GTPases involved in the control of actin cytoskeleton dynamics, proliferation and survival. Rnd1, Rnd2 and RhoE/Rnd3 form a subfamily of Rho proteins characterized by being constitutively active. The role of these proteins has been studied during the last years in several systems; however, little is known about their expression and functions in the reproductive organs. In this work we analysed the localization and the effect of RhoE deficiency in the testes using mice lacking RhoE expression (RhoE gt/gt), and our research shows some unexpected and relevant results. First, we have observed that RhoE is only expressed in Leydig cells within the testicular parenchyma and it is absent of seminiferous tubules. In addition, RhoE is expressed in the excurrent ducts of the testis, including the ductuli efferentes, epididymis and ductus deferens. Moreover, the testes of postnatal 15-day-old RhoE null mice are smaller, both in absolute values and in relation to the body weight. Furthermore, the dimensions of their seminiferous tubules are also reduced compared with wild-types. In order to study the role of RhoE in the adult, we analysed heterozygous animals as RhoE null mice die early postnatally. Our results show that the testes of adult RhoE heterozygous mice are also smaller than those of the wild-types, with a 17% decrease in the ratio testis weight/body weight. In addition, their seminiferous tubules have reduced tubular diameter (12%) and a thinner epithelial wall (33%) that appears disorganized and with a swollen lumen. Finally, and probably as a consequence of those alterations, the sperm concentration of heterozygous animals was found to be lower than in the wild-types. These results indicate that accurate levels of RhoE in the testes are necessary for a correct development and function of male gonads, and suggest novel and unexpected roles of Rnd GTPases in the reproductive physiology.

The use of blended learning strategies is increasingly common in health sciences, including veterinary medicine; however, there are very few descriptions of these methods being applied to practicals. We describe here the application of blended learning based on the implementation of flipped classrooms with collaborative learning and gamification to the 2020–2021 veterinary medicine gross anatomy practicals at CEU Cardenal Herrera University (Spain). Students prepared for the sessions by pre-viewing videos and taking a quiz before the start. The sessions were conducted in small groups where students learned through collaborative work and reviewed their learning with a card game. A small but significant increase was observed when comparing the scores of practical exams of the locomotor apparatus with those of 2018–2019 (6.79 ± 2.22 vs. 6.38 ± 2.24, p < 0.05), while the scores were similar (7.76 ± 1.99 vs. 7.64 ± 1.92) for the organ system exams. Students’ responses in a satisfaction survey were mostly positive (>80%) regarding the motivating and learning-facilitating effect of this educational method. Our work shows that the application of blended learning in anatomy practicals based on a flipped classroom and with elements of gamification and collaborative work can be an effective way to improve the learning experience of students.

The use of cadavers is essential for veterinary anatomy learning. However, facing an animal corpse can be stressful for veterinary students because of their empathy toward animals. The objective of this study was to evaluate veterinary medicine students' emotions, feelings, and anxiety levels related to practicals with dog cadavers. Two questionnaires were administered to 1st year students (n = 168) at CEU Cardenal Herrera University in Valencia (Spain) before and after their first practical session with cadavers. The application of State–Trait Anxiety Inventory questionnaires showed that “state anxiety” decreased significantly (p < 0.05), from a score of 14.8 before the practical to 10.4 after, and that female students showed higher but not significantly different levels than males. Most (64%) of the students were not willing to donate the bodies of their pets, and those students were more stressed before the practical than their peers, although their anxiety levels significantly decreased by the end of the session. The majority of the students answered positively about emotions, such as feeling calm, safe, not nervous, relaxed and not worried before the practical, and this increased significantly to more than 80% by the end of the session. The visualization of educational videos prior to the session was evaluated positively by students. These results agree with those reported in other health science disciplines, showing that students face practical sessions with corpses in a similar way and suggesting that the use of videos can help decrease anxiety and enhance their learning experience.

Ultrasound-guided quadratus lumborum block (QLB) is a locoregional technique described in canine cadavers. The aim of this study was to assess a modified approach to QLB to minimise potential complications such as abdominal organ puncture. Nine canine cadavers were included and were positioned in lateral recumbency. An ultrasound-guided QLB was performed on each side. The probe was placed in the transverse position over the lumbar muscles just caudal to the last rib, and a needle was advanced in-plane from a dorso-lateral to a ventro-medial. A volume of 0.2 mL kg􀀀1 of a mixture of iomeprol and methylene blue was injected. Computed tomography (CT) and dissection were performed to evaluate the spreading. Success was defined as staining of the nerve with a length of more than 0.6 cm. Potential complications such as intra-abdominal, epidural, or intravascular spreading of the mixture were also assessed. The CT images showed a T13 to L7 vertebra distribution, with a median of 5 (3–6). Dissection showed staining of the nerves from T13 to L4, with a median of 3 (2–5). No complications were found. This modified approach to QLB is safe and shows similar results to the previous studies in canine carcass.