The immunological synapse (IS), a highly organized structure that forms at the point of contact between a T cell and an APC, is essential for the proper development of signaling events, including the Ca²⁺ response. Kv1.3 channels control Ca²⁺ homeostasis in human T cells and move into the IS upon Ag presentation. However, the process involved in channel accumulation in the IS and the functional implications of this localization are not yet known. Here we define the movement of Kv1.3 into the IS and study whether Kv1.3 localization into the IS influences Ca²⁺ signaling in Jurkat T cells. Crosslinking of the channel protein with an extracellular Ab limits Kv1.3 mobility and accumulation at the IS. Moreover, Kv1.3 recruitment to the IS does not involve the transport of newly synthesized channels and it does not occur through recycling of membrane channels. Kv1.3 localization in the IS modulates the Ca²⁺ response. Blockade of Kv1.3 movement into the IS by crosslinking significantly increases the amplitude of the Ca²⁺ response triggered by anti-CD3/anti-CD28-coated beads, which induce the formation of the IS. On the contrary, the Ca²⁺ response induced by TCR stimulation without the formation of the IS with soluble anti-CD3/anti-CD28 Abs is unaltered. The results presented herein indicate that, upon Ag presentation, membrane-incorporated Kv1.3 channels move along the plasma membrane to localize in the IS. This localization is important to control the amplitude of the Ca²⁺ response, and disruption of this process can account for alterations of downstream Ca²⁺-dependent signaling events.

Aberrant T cell responses during T cell activation and immunological synapse (IS) formation have been described in systemic lupus erythematosus (SLE). Kv1.3 potassium channels are expressed in T cells where they compartmentalize at the IS and play a key role in T cell activation by modulating Ca²⁺ influx. Although Kv1.3 channels have such an important role in T cell function, their potential involvement in the etiology and progression of SLE remains unknown. This study compares the K channel phenotype and the dynamics of Kv1.3 compartmentalization in the IS of normal and SLE human T cells. IS formation was induced by 1–30 min exposure to either anti-CD3/CD28 Ab-coated beads or EBV-infected B cells. We found that although the level of Kv1.3 channel expression and their activity in SLE T cells is similar to normal resting T cells, the kinetics of Kv1.3 compartmentalization in the IS are markedly different. In healthy resting T cells, Kv1.3 channels are progressively recruited and maintained in the IS for at least 30 min from synapse formation. In contrast, SLE, but not rheumatoid arthritis, T cells show faster kinetics with maximum Kv1.3 recruitment at 1 min and movement out of the IS by 15 min after activation. These kinetics resemble preactivated healthy T cells, but the K channel phenotype of SLE T cells is identical to resting T cells, where Kv1.3 constitutes the dominant K conductance. The defective temporal and spatial Kv1.3 distribution that we observed may contribute to the abnormal functions of SLE T cells.

T cell receptor engagement results in the reorganization of intracellular and membrane proteins at the T cell-antigen presenting cell interface forming the immunological synapse (IS), an event required for Ca²⁺ influx. KCa3.1 channels modulate Ca²⁺ signaling in activated T cells by regulating the membrane potential. Nothing is known regarding KCa3.1 membrane distribution during T cell activation. Herein, we determined whether KCa3.1 translocates to the IS in human T cells using YFP-tagged KCa3.1 channels. These channels showed electrophysiological and pharmacological properties identical to wild-type channels. IS formation was induced by either anti-CD3/CD28 antibody-coated beads for fixed microscopy experiments or Epstein-Barr virus-infected B cells for fixed and live cell microscopy. In fixed microscopy experiments, T cells were also immunolabeled for F-actin or CD3ε, which served as IS formation markers. The distribution of KCa3.1 was determined with confocal and fluorescence microscopy. We found that, upon T cell activation, KCa3.1 channels localize with F-actin and CD3ε to the IS but remain evenly distributed on the cell membrane when no stimulus is provided. Detailed imaging experiments indicated that KCa3.1 channels are recruited in the IS shortly after antigen presentation and are maintained there for at least 15–30 min. Interestingly, pretreatment of activated T cells with the specific KCa3.1 blocker TRAM-34 blocked Ca²⁺ influx, but channel redistribution to the IS was not prevented. These results indicate that KCa3.1 channels are a part of the signaling complex that forms at the IS upon antigen presentation.

The integration of artificial intelligence (AI) into the biological sciences marks a transformative era, reshaping research methodologies, data analysis, and hypothesis generation. This technological advancement accelerates discoveries and enhances our understanding of complex biological systems. As AI increasingly influences decision-making processes, the necessity for students and scientists to critically assess AI-generated outputs becomes paramount. The current narrative review explores the evolving role of critical thinking in biological sciences amidst the rise of AI, emphasizing the importance of skepticism, contextual understanding, and ethical considerations. It argues that while AI provides powerful tools for data interpretation and pattern recognition, human oversight and critical analysis remain indispensable to validate findings and prevent biases inherent in automated systems. Higher education institutions play a crucial role in fostering a culture of critical thinking, equipping biological scientists to effectively harness AI technologies while ensuring the integrity of their research and upholding scientific and ethical standards. Furthermore, AI tools, including chatbots, could be strategically employed in active learning methodologies, such as problem-based learning, flipped classrooms, and online learning. These methodologies enhance the ability of students to effectively utilize AI technologies while ensuring the rigor of scientific research. In conclusion, the current review underscores the benefits, challenges, and educational implications of AI integration, offering actionable insights for educators and learners seeking to adapt effectively to this rapidly evolving technological landscape.

Microbial physiology is a basic course taught throughout biomedical science disciplines. Students study the structure, growth, and metabolism of microorganisms and often find it difficult to learn the information, usually because they fail to see the wider applications. The current microbiology laboratory series describes how to transform a “cookbook” undergraduate laboratory to an inquiry-based one by incorporating problem-based learning. The students use a food poisoning case study that develops over a series of seven experiments and take on the role of the microbiology technician who is responsible for coming up with the answer and submitting a report to a clinician. The case provides coherence to the sessions, and the students are given the opportunity to learn about, and practice, common techniques they would encounter in a clinical microbiology laboratory. Those include the aseptic method, cultivation of bacteria, quantification of bacteria in culture, isolation of pure culture, morphological observation by light microscopy, Gram staining, the use of selective and differential media, and the effectiveness of a variety of antimicrobials and antibiotics. This laboratory series has been designed so that it can be implemented in any setting, using simple materials and inexpensive, nonspecialized equipment. The experiments are carried out in small groups, and a facilitator may tutor up to two groups of 10 students at a time. The current method has been successfully implemented for the past 2 yr, and the students demonstrated greater motivation in learning and understanding.