Laura Cecilia Giojalas

Molecular Cell Biology (MCB) should be taught according to the scientific practices, avoiding cumulative and memory knowledge construction, but favoring scientific thinking. A way to achieve this goal is to apply activities involving scientific news, which construct knowledge through significant learning and the development of critical thinking. The study aimed to evaluate the implementation of learning activities involving scientific news in the MCB course at the undergraduate level. The perspective of design‐based research was applied, whereas the cognitive and interpersonal aspects were evaluated by means of the professors' narration, class registration, questionnaires answered by the students, and the evaluation of the activities carried out by the students. Results showed that the activity involving scientific news reinforces the construction and integration of new knowledge with that previously acquired and consolidates the acquisition of scientific thinking. Even though the completion of the activity involved a complex process, according to the professor and students' opinion, the students observed positive aspects such as the application of biological and scientific language and the motivation to search for related information. Regarding the cooperative learning strategy, students perceived that it is a methodology that facilitates their learning. In summary, incorporating scientific news into MCB courses will enhance professors' effectiveness in achieving didactic goals while also fostering scientific thinking in students, equipping them for future roles as biologists and professors in biological sciences.

The sperm selection assay (SSA) is a method based on chemotaxis to obtain spermatozoa at the optimum physiological state to successfully fertilize the egg. It consists of a device made of acrylic and an attractant solution which includes progesterone. We evaluate potential cytotoxicity interactions by means of Neutral Red uptake, the MTT and colony formation assays, according to ISO normative. Here we showed that even stressing the conditions of the assays, the SSA device alone or together with the progesterone solutions employed, showed to be innocuous for the cells. Suggesting that SSA could be incorporated into the ART procedures.

Nowadays Molecular Cell Biology (MCB) must be taught as science is practiced. Even though there are several approaches based on scientific practices, a key aspect is to define the purpose of each of these teaching strategies and, most importantly, their implementation. Our goal was to train students to acquire, understand, and communicate new scientific knowledge in the field. The main feature of our new teaching methodology was progressive training in scientific practices associated with a back‐and‐forward interplay between activities and assessments. The methodology was implemented over 4 years, in students attending the MCB course of the undergraduate degree in Biological Sciences. In the first two modules, the students were prepared to comprehend MCB concepts and techniques and to experience activities based on scientific practices. In the third module, the students analyzed a primary paper in‐depth. They were assessed by midterm exams based on a primary paper, written laboratory reports, and the oral presentation of a scientific paper. Our teaching proposal was evaluated through the students' academic performance and by their opinion on the teaching methodology. Most students were satisfied since they improved their acquisition of concepts, their interpretation and integration of scientific knowledge, and developed skills to communicate scientific knowledge in writing and orally. The novelty of transversal interconnections and progressive training in scientific practices provides students with skills in acquiring and understanding new scientific information, even beyond the MCB course.

Sperm chemotaxis may facilitate the finding of the oocyte. Only capacitated spermatozoa can orient their movement by chemotaxis, which as well as capacitation, is regulated in part by the cAMP-PKA pathway. Reactive oxygen species (ROS) are produced during sperm capacitation which is closely related to chemotaxis. Then, the ROS participation in the chemotactic signaling can be expected. Here we studied the role of ROS in the chemotaxis signaling of equine spermatozoa which produce high quantities of ROS because of their energy metabolism. The level of capacitated and chemotactic spermatozoa was increased with 0.1 and 0.2 mM hydrogen peroxide (H₂O₂), which was involved in the chemotactic signaling. By combining a concentration gradient of H₂O₂ with inhibitors/chelators of some of the signaling pathway elements, we showed that the activation of NOX (membrane NADPH oxidase) increases the intracellular ROS which activate the chemotaxis AMPc-PKA pathway. Our results provide evidence about the participation of ROS in the chemotactic signaling mediated by progesterone (P).

Infertility is a common medical condition encountered by health systems throughout the world. Despite the development of complex in vitro fertilization techniques, only one-third of these procedures are successful. New lab-on-a-chip systems that focus on spermatozoa selection require a better understanding of sperm behavior under ultra-confined conditions in order to improve outcomes. Experimental studies combined with models and simulations allow the evaluation of the efficiency of different lab-on-a-chip devices during the design process. In this work, we provide experimental evidence of the dynamics of sperm interacting with a lateral wall in a shallow chamber. We observe a decrease in average sperm velocity during initial wall interaction and partial recovery after the alignment of the trajectory of the cell. To describe this phenomenon, we propose a simple model for the sperm alignment process with a single free parameter. By incorporating experimental motility characterization into the model, we achieve an accurate description of the average velocity behavior of the sperm population close to walls. These results will contribute to the design of more efficient lab-on-a-chip devices for the treatment of human infertility.Infertility is a common medical condition encountered by health systems throughout the world. Despite the development of complex in vitro fertilization techniques, only one-third of these procedures are successful. New lab-on-a-chip systems that focus on spermatozoa selection require a better understanding of sperm behavior under ultra-confined conditions in order to improve outcomes. Experimental studies combined with models and simulations allow the evaluation of the efficiency of different lab-on-a-chip devices during the design process. In this work, we provide experimental evidence of the dynamics of sperm interacting with a lateral wall in a shallow chamber. We observe a decrease in average sperm velocity during initial wall interaction and partial recovery after the alignment of the trajectory of the cell. To describe this phenomenon, we propose a simple model for the sperm alignment process with a single free parameter. By incorporating experimental motility characterization into the model, we ach...

Infertility is a common medical condition encountered by health systems throughout the world. Despite the development of complex in vitro fertilization techniques, only one-third of these procedures are successful. New lab-on-a-chip systems that focus on spermatozoa selection require a better understanding of sperm behavior under ultra-confined conditions in order to improve outcomes. Experimental studies combined with models and simulations allow the evaluation of the efficiency of different lab-on-a-chip devices during the design process. In this work, we provide experimental evidence of the dynamics of sperm interacting with a lateral wall in a shallow chamber. We observe a decrease in average sperm velocity during initial wall interaction and partial recovery after the alignment of the trajectory of the cell. To describe this phenomenon, we propose a simple model for the sperm alignment process with a single free parameter. By incorporating experimental motility characterization into the model, we achieve an accurate description of the average velocity behavior of the sperm population close to walls. These results will contribute to the design of more efficient lab-on-a-chip devices for the treatment of human infertility.

Cells can communicate with other neighboring or distant cells through the secretion of extracellular vesicles (EV), composed of a lipid bilayer and bearing surface molecules that allow them to recognize target cells. In this way, EV induce signaling via different mechanisms, modulating the physiological state of the recipient cell. EV have been identified in both male and female reproductive fluids, however, the possible role of EV isolated from female reproductive fluids has become an emerging field only recently. It is known that ejaculated mammalian spermatozoa need to undergo physiological preparation in the female reproductive tract to fertilize the egg. EV secreted by different regions of the female tract constitute signals that may have a key role in regulating sperm functions. The aims of the present study were isolating EV from different regions of the bovine oviduct and analyzing their interaction and physiological effects on spermatozoa. Here, we report the characterization of bovine oviductal fluid EV from the isthmus and ampulla region and their effect on the induced acrosome reaction and signaling events associated with sperm capacitation. EV induced an increase in sperm protein tyrosine phosphorylation, while cell survival of cryopreserved bovine spermatozoa was maintained. We also show that EV uptake regulates the sperm calcium levels by inducing an immediate increase in the intracellular calcium concentration and sperm priming, after a pre‐incubation period, of the progesterone‐induced intracellular calcium rise. Our data contribute to understand the role of EV in the communication between the female reproductive tract and the sperm physiology, information that may be used to improve the efficiency of reproductive assisted technologies.

Human sperm show a variety of different behaviours (types of motility) that have different functional roles. Previous reports suggest that sperm may reversibly switch between these behaviours. We have recorded and analysed the behaviour of individual human sperm (180 cells in total), each cell monitored continuously for 3–3.5 min either under control conditions or in the presence of Ca²⁺-mobilising stimuli. Switching between different behaviours was assessed visually (1 s bins using four behaviour categories), and was verified by fractal dimension analysis of sperm head tracks. In the absence of stimuli, ~90% of cells showed at least one behavioural transition (mean rate under control conditions = 6.4 ± 0.8 transitions.min⁻¹). Type 1 behaviour (progressive, activated-like motility) was most common, but the majority of cells (>70%) displayed at least three behaviour types. Treatment of sperm with Ca²⁺-mobilising agonists had negligible effects on the rate of switching but increased the time spent in type 2 and type 3 (hyperactivation-like) behaviours (P < 2^*10⁻⁸; chi-square). Treatment with 4-aminopyridine under alkaline conditions (pH_o = 8.5), a highly-potent Ca²⁺-mobilising stimulus, was the most effective in increasing the proportion of type 3 behaviour, biasing switching away from type 1 (P < 0.005) and dramatically extending the duration of type 3 events (P < 10⁻¹⁶). Other stimuli, including 300 nM progesterone and 1% human follicular fluid, had qualitatively similar effects but were less potent. We conclude that human sperm observed in vitro constitutively display a range of behaviours and regulation of motility by [Ca²⁺]_i, at the level of the single cell, is achieved not by causing cells to adopt a ‘new’ behaviour but by changing the relative contributions of those behaviours.