Tubule Cells Research Articles

IL-22 is secreted by proximal tubule cells and regulates DNA damage response and cell death in acute kidney injury

Acute kidney injury (AKI) affects over 13 million people worldwide annually and is associated with a 4-fold increase in mortality. Our lab and others have shown that DNA damage response (DDR) governs the outcome of AKI in a bimodal manner. Activation of DDR sensor kinases protects against AKI, while hyperactivation of DDR effector proteins, such as p53, induces cell death and worsens AKI. The factors that trigger DDR to switch from pro-repair to pro-cell death remain to be resolved. Here we investigated the role of interleukin 22 (IL-22), an IL-10 family member whose receptor (IL-22RA1) is expressed on proximal tubule cells (PTCs), in DDR activation and AKI. Using cisplatin and aristolochic acid (AA) induced nephropathy as models of DNA damage, we identified PTCs as a novel source of urinary IL-22. Functionally, IL-22 binding IL-22RA1 on PTCs amplified the DDR. Treating primary PTCs with IL-22 alone induced rapid activation of the DDR. The combination of IL-22 and either cisplatin- or AA-induced cell death in primary PTCs, while the same dose of cisplatin or AA alone did not. Global deletion of IL-22 protected against cisplatin- or AA-induced AKI, reduced expression of DDR components, and inhibited PTC cell death. To confirm PTC IL-22 signaling contributed to AKI, we knocked out IL-22RA1 specifically in kidney tubule cells. IL-22RA1ΔTub mice displayed reduced DDR activation, cell death, and kidney injury compared to controls. Thus, targeting IL-22 represents a novel therapeutic approach to prevent the negative consequences of the DDR activation while not interfering with repair of damaged DNA.

Susceptibility of the cultured Amazonian fish, Colossoma macropomum, to experimental infection with Aeromonas species from ornamental fish

The global ornamental fish trade carries important risk factors for spreading pathogens between different countries and regions, not only for ornamental fish but also for cultured fish and even other animal species. In the current study, we reported the capacity of Aeromonas veronii and A. hydrophila isolated from ornamental fish to experimentally infect the reared Amazonian fish Colossoma macropomum. For this, those bacteria were identified, and a primary characterization was performed. Fish were inoculated with 0.1 mL of increasing concentrations of A. hydrophila or A. veronii (C1 = 1 × 102; C2 = 1.8 × 104; C3 = 2.1 × 106; C4 = 2.4 × 108 bacterial cells per mL) in the coelomic cavity. In the control group, fish received the same volume of sterile saline solution (0.9 %). Fish presented petechiae, skin suffusions, and mortality rates up to 100 % according to the inoculum concentration. Histopathologically, fish presented necrosis with karyolysis, loss of the cytoplasmic delimitation of cells of the renal tubules and hepatocytes, hemorrhage, cellular edema, and the presence of bacterial cells. The LD50–96h of A. veronii on C. macropomum was estimated at 2.4 × 106 CFU mL−1 and of A. hydrophila at 1.408 × 105 CFU mL−1. The results demonstrated that it is possible that Aeromonas species isolated from ornamental fish affect C. macropomum, causing similar clinical signs and lesions. This shows the importance of promoting risk control measures worldwide regarding the trade of ornamental fish.

The Effect of Dosing with Lemon Juice on the Renal Tissue of Laboratory Rats with Kidney Stones (Urolithiasis/ Nephrolithiasis) Experimentally

Kidney stones and renal tissue damage are common diseases that are affected by many causes, including nutritional causes. There have been many ways to treat kidney disease and ways to prevent it through appropriate diets. The research aimed to study the effectiveness of lemon juice in preventing kidney disease in laboratory rats. As 15 animals were distributed in three groups, the first was considered a control group that was dosed with distilled water and the other group was dosed with ethylene glycol as a cause of kidney injury, while lemon juice was used to treat the latter. It was found through the histological sections of the kidneys that the lemon extract helped to preserve the kidneys of the animals, as the glomeruli appeared regularly with the integrity of the epithelial cells of the renal tubules. It was found that the rates of damage to the renal glomeruli in the group treated with lemon did not differ significantly from the animals that were not exposed to infection (the control group). And the damage rate is about 22%. The percentage of damage to the glomeruli in the histological sections of animals that were dosed with the causative agent of stone formation (ethylene glycol) and not treated was about 75%, as the results of histological cutting indicated that they contained atrophy in the renal glomeruli with dissection of Bowman’s capsule and hemorrhage on the surface with calcium crystals deposits.

Granulocyte-macrophage colony-stimulating factor (GM-CSF)-induced maturation of spermatogonial cells from prepubertal mice in vitro is enhanced by testosterone.

Spermatogenesis is the complicated process of sperm generation. During this process, spermatogonial cells proliferate and differentiate via meiotic and post-meiotic stages to produce mature sperm. This process is under the regulation of testicular autocrine/paracrine factors. In addition, endocrine factors are crucial to complete spermatogenesis. We aimed to localize granulocyte-macrophage colony-stimulating factor (GM-CSF) and its receptor (GM-CSFR) in testicular cells and further evaluate its involvement in the development of spermatogenesis in vitro. Weisolated cells from seminiferous tubule cells of seven-day-old mice and cultured them in vitro using a methylcellulose culture system (MCS), in the presence of GM-CSF and/or testosterone for four weeks. The cells were then examined for markers of different stages of spermatogenesis by immunofluorescence staining and/or qPCR analyses. Our results revealed the presence of GM-CSF and GM-CSFR in testicular cells (premeiotic and meiotic cells as well as somatic cells; Leydig and Sertoli cells). We further demonstrated the development of colonies/spheroids in the MCS which contained pre-meiotic, meiotic, and post-meiotic cells. The addition of GM-CSF to the MCS significantly increased the percentage of pre-meiotic and meiotic cells compared to control. Furthermore, the addition of GM-CSF and testosterone together significantly increased the percentage of cells in the post-meiotic stage compared to the addition of each separately. In conclusion, our results indicate that testicular cells express GM-CSF/GM-CSFR, and that GM-CSF is involved in the development of different stages of spermatogenesis in vitro. Furthermore, testosterone enhances the development of spermatogenic cells and potentiates the effect of GMCSF on the development of post-meiotic cells. These findings provide evidence that GM-CSF and testosterone are involved in the development of spermatogenesis in vitro and in vivo. In brief: Testicular somatic and germ cells express GM-CSF and GM-CSFR. Our study suggests that testicular GM-CSF is involved in the development of spermatogenesis, which is potentiated by testosterone.

High-Glucose-Induced Injury to Proximal Tubules of the Renal System Is Alleviated by Netrin-1 Suppression of Akt/mTOR.

The kidneys have a high level of Netrin-1 expression, which protects against some acute and chronic kidney disorders. However, it is yet unknown how Netrin-1 affects renal proximal tubule cells in diabetic nephropathy (DN) under pathological circumstances. Research has shown that autophagy protects the kidneys in animal models of renal disease. In this study, we looked at the probable autophagy regulation mechanism of Netrin-1 and its function in the pathogenesis of DN. We proved that in HK-2 cell, high blood sugar levels caused Netrin-1 to be downregulated, which then triggered the Akt/mTOR signaling pathway and enhanced cell death and actin cytoskeleton disruption. By adding Netrin-1 or an autophagy activator in vitro, these pathogenic alterations were reverted. Our results indicate that Netrin-1 stimulates autophagy by blocking the Akt/mTOR signaling pathway, which underlies high-glucose-induced malfunction of the renal proximal tubules. After HK-2 cells were incubated with Netrin-1 recombination protein and rapamycin under HG conditions for 24 h, the apoptosis was significantly reduced, as shown by the higher levels of Bcl-2, as well as lower levels of Bax and cleaved caspase-3 (P = 0.012, Cohen's d = 0.489, Glass's delta = 0.23, Hedges' g = 0.641). This study reveals that targeting Netrin-1-related signaling has therapeutic potential for DN and advances our knowledge of the processes operating in renal proximal tubules in DN.

MiR-125a-5p/miR-125b-5p contributes to pathological activation of angiotensin II-AT1R in mouse distal convoluted tubule cells by the suppression of Atrap

The renin-angiotensin system plays a crucial role in the regulation of blood pressure. Activation of the angiotensin II (Ang II)-Ang II type 1 receptor (AT1R) signaling pathway contributes to the pathogenesis of hypertension and subsequent organ damage. AT1R-associated protein (ATRAP) has been identified as an endogenous inhibitory protein of the AT1R pathological activation. We have shown that mouse Atrap (Atrap) represses various Ang II-AT1R-mediated pathologies, including hypertension in mice. The expression of human ATRAP (ATRAP)/Atrap can be altered in various pathological states in humans and mice, such as Ang II stimulation and serum starvation. However, the regulatory mechanisms of ATRAP/Atrap are not yet fully elucidated. miRNAs are 21 to 23 nucleotides of small RNAs that post-transcriptionally repress gene expression. Single miRNA can act on hundreds of target mRNAs, and numerous miRNAs have been identified as the Ang II-AT1R signaling-associated disease phenotype modulator, but nothing is known about the regulation of ATRAP/Atrap. In the present study, we identified miR-125a-5p/miR-125b-5p as the evolutionarily conserved miRNAs that potentially act on ATRAP/Atrap mRNA. Further analysis revealed that miR-125a-5p/miR-125b-5p can directly repress both ATRAP and Atrap. In addition, the inhibition of miR-125a-5p/miR-125b-5p resulted in the suppression of the Ang II-AT1R signaling in mouse distal convoluted tubule cells. Taken together, miR-125a-5p/miR-125b-5p activates Ang II-AT1R signaling by the suppression of ATRAP/Atrap. Our results provide new insights into the potential approaches for achieving the organ-protective effects by the repression of the miR-125 family associated with the enhancement of ATRAP/Atrap expression.

The Effect of Fermented Honey on Seminiferous Tubule Cells While being Induced by Cisplatin and Islamic Perspective Review

Background: Cisplatin is the most effective cancer treatment used in chemotherapy. Cisplatin works by inducing oxidative stress, but it also has toxic side effects on healthy cells in the male reproductive organs. Therefore, there is a need for a solution to reduce the side effects of cisplatin. Honey fermentation is a natural substance that can reduce the toxicity of cisplatin. Honey is known as “The food of God” which is a gift of sustenance from Allah SWT due to its extraordinary benefits. Methodology: This study utilized primary data with an experimental posttest-only control group design conducted in vivo. The research involved four experimental groups of rats: a control group without any treatment, a cisplatin group, a 5% honey fermentation group exposed to cisplatin, and a 10% honey fermentation group exposed to cisplatin. All rats were weighed before, during, and after the treatment. Subsequently, HE staining was performed for both descriptive and quantitative observations. Results: Honey fermentation can provide a protective effect on primary spermatocytes and Leydig interstitial cells, almost reaching normal levels. The highest improvement was observed in the 5% honey fermentation group for primary spermatocytes and the 10% group for Leydig interstitial cells. Conclusion: Honey fermentation administration can be protective for testicular organ cells against the side effects of cisplatin. From an Islamic perspective, Allah has created honey for consumption, as mentioned in Surah An-Nahl.

Pax protein depletion in proximal tubules triggers conserved mechanisms of resistance to acute ischemic kidney injury preventing transition to chronic kidney disease

Acute kidney injury (AKI) is a common condition that lacks effective treatments. In part, this shortcoming is due to an incomplete understanding of the genetic mechanisms that control pathogenesis and recovery. Identifying the molecular and genetic regulators unique to nephron segments that dictate vulnerability to injury and regenerative potential could lead to new therapeutic targets to treat ischemic kidney injury. Pax2 and Pax8 are homologous transcription factors with overlapping functions that are critical for kidney development and are re-activated in AKI. Here, we examined the role of Pax2 and Pax8 in recovery from ischemic AKI and found them upregulated after severe AKI and correlated with chronic injury. Surprisingly, proximal-tubule-selective deletion of Pax2 and Pax8 resulted in a less severe chronic injury phenotype. This effect was mediated by protection against the acute insult, similar to pre-conditioning. Prior to injury, Pax2 and Pax8 mutant mice develop a unique subpopulation of proximal tubule cells in the S3 segment that displayed features usually seen only in acute or chronic injury. The expression signature of these cells was strongly enriched with genes associated with other mechanisms of protection against ischemic AKI including caloric restriction, hypoxic pre-conditioning, and female sex. Thus, our results identified a novel role for Pax2 and Pax8 in mature proximal tubules that regulates critical genes and pathways involved in both the injury response and protection from ischemic AKI.

Significance of Organic Anion Transporter 2 and Organic Cation Transporter 2 in Creatinine Clearance: Mechanistic Evaluation Using Freshly Prepared Human Primary Renal Proximal Tubule Cells.

Creatinine, a clinical marker for kidney function, is predominantly cleared by glomerular filtration, with active tubular secretion contributing to about 30% of its renal clearance. Recent studies suggested the potential involvement of organic anion transporter (OAT)2, in addition to the previously known organic cation transporter (OCT)2-mediated basolateral uptake, in creatinine active secretion. Here we characterized the transport mechanisms of creatinine using transfected human embryonic kidney (HEK)293 cells and freshly prepared human primary renal proximal tubule epithelial cells (hPTCs). Creatinine showed transport by OAT2 in transfected HEK293 cells. In addition, both creatinine and metformin showed transport by OCT2 and multidrug and toxin extrusion pump (MATE)1 and MATE2K, while penciclovir was selective for OAT2. Time-dependent cell accumulation was observed for creatinine and metformin in hPTCs. Their accumulation was increased by pyrimethamine but inhibited by decynium-22, likely due to differential inhibition of OCT2 versus MATEs. Additionally, indomethacin (an OAT2 inhibitor) reduced penciclovir uptake (∼75%) in hPTCs illustrating functional OAT2 activity. However, no modulation of creatinine and metformin cell accumulation was apparent with indomethacin. Creatinine transport characteristics in the presence of inhibitors approached those of metformin, an OCT2/MATE substrate, but were distinct from those of penciclovir, an OAT2-selective substrate. Moreover, indomethacin showed no significant effect on the basolateral-to-apical transport and net secretion of creatinine across hPTC monolayers. Collectively, the functional studies suggest OCT2 as the primary basolateral uptake mechanism and that OAT2 has a minimal role, in creatinine renal secretion. Our results highlight the utility of hPTCs to enable the functional assessment of renal transport mechanisms. SIGNIFICANCE STATEMENT: Our results obtained with primary hPTCs indicate that OCT2/MATE (vs. OAT2) play a major role in the active renal secretion of creatinine. Quantitative pharmacokinetic models should therefore focus on OCT2/MATE when describing serum creatinine and creatinine clearance modulation by inhibitor drugs and genotype- or disease-related activity changes. The present study highlights the utility of freshly isolated hPTCs to support solute carrier phenotyping to enable the functional assessment of renal transport mechanisms.

Reactive Blue 2 Labels Protamine in Late-Haploid Spermatids and Spermatozoa and Can Be Used for Toxicity Evaluation

Reactive blue 2 (RB2) dye specifically binds to the nuclei of human spermatozoa under weakly alkaline conditions, thereby providing a new method for assessing sperm quality. However, this technique has not yet been applied to other mammalian species, such as well-established rodent models, which would allow evaluation of the male reproductive toxicity of new drug candidates in nonclinical studies. We aimed to evaluate the usefulness of RB2 staining in assessing testicular and epididymal sperm toxicity in mice using a busulfan-induced infertility model. Male C57BL/6J mice were intraperitoneally administered 40 mg/kg of busulfan. After 28 days, the testes and epididymis were collected and stained with RB2 at pH 10. In vitro evaluations were conducted on uncoated glass slides with RB2 mixed with mouse synthetic protamines, protamines extracted from the human spermatozoa or intracellular protein components from somatic cells without protamines. Following peanut agglutinin lectin histochemistry, RB2-positive cells were observed in elongating and elongated spermatids at all stages except for stages IX–XI of the seminiferous epithelium. After busulfan administration, the proportion of RB2-positive germ cells in the seminiferous tubules was significantly decreased, and no RB2-positive spermatozoa were found in the caput epididymis of treated mice. Aggregates were observed in a mixture of RB2 dye (pH 10) and protamines but not in a mixture of intracellular protein components without protamines, and this specificity was lost at a neutral pH. Our study demonstrated that RB2 specifically stains steps 12–16 spermatids, indicating specific binding to the protamines expressed in these spermatids. The RB2 staining technique has potential as a biomarker for male reproductive toxicity, allowing for the rapid visualization of protamination in an animal model commonly used for the evaluation of male reproductive toxicity.

IL-22 Secreted from Proximal Tubule Cells Regulates Cell Fate During AKI

IL-22 Secreted from Proximal Tubule Cells Regulates Cell Fate During AKI

Proteomic Profiling of a Novel Immortalised Human Distal Convoluted Tubule Cell Line

Proteomic Profiling of a Novel Immortalised Human Distal Convoluted Tubule Cell Line

The SGLT2 inhibitor dapagliflozin improves kidney function in glycogen storage disease XI.

Glycogen storage disease XI, also known as Fanconi-Bickel syndrome (FBS), is a rare autosomal recessive disorder caused by mutations in the SLC2A2 gene that encodes the glucose-facilitated transporter type 2 (GLUT2). Patients develop a life-threatening renal proximal tubule dysfunction for which no treatment is available apart from electrolyte replacement. To investigate the renal pathogenesis of FBS, SLC2A2 expression was ablated in mouse kidney and HK-2 proximal tubule cells. GLUT2Pax8Cre+ mice developed time-dependent glycogen accumulation in proximal tubule cells and recapitulated the renal Fanconi phenotype seen in patients. In vitro suppression of GLUT2 impaired lysosomal autophagy as shown by transcriptomic and biochemical analysis. However, this effect was reversed by exposure to a low glucose concentration, suggesting that GLUT2 facilitates the homeostasis of key cellular pathways in proximal tubule cells by preventing glucose toxicity. To investigate whether targeting proximal tubule glucose influx can limit glycogen accumulation and correct symptoms in vivo, we treated mice with the selective SGLT2 inhibitor dapagliflozin. Dapagliflozin reduced glycogen accumulation and improved metabolic acidosis and phosphaturia in the animals by normalizing the expression of Napi2a and NHE3 transporters. In addition, in a patient with FBS, dapagliflozin was safe, improved serum potassium and phosphate concentrations, and reduced glycogen content in urinary shed cells. Overall, this study provides proof of concept for dapagliflozin as a potentially suitable therapy for FBS.

Early Transcriptional Changes in Distal Convoluted Tubule Cells Are Evident in PKD1 (Polycystin 1) Mutant Mice Prior to Cyst Development

Early Transcriptional Changes in Distal Convoluted Tubule Cells Are Evident in PKD1 (Polycystin 1) Mutant Mice Prior to Cyst Development

Regulation of the Adaptive Proximal Tubule Cell State by ELF3, KLF6, and KLF10

Regulation of the Adaptive Proximal Tubule Cell State by ELF3, KLF6, and KLF10

Morphofunctional state of the kidneys of laboratory rats during acute respiratory distress syndrome

One of the most common complications of acute respiratory distress syndrome is acute kidney injury, the mechanisms of development of which remain not completely clarified. The purpose of this study is to examine morphofunctional changes in the kidneys of rats with induced acute respiratory distress syndrome at various time intervals after modelling the pathology. For the study, 56 healthy sexually mature male rats weighing 200-220 g were used, divided into 7 groups: control, 6, 24 hours, 3, 7, 14, and 28 days of the experiment. Respiratory distress in animals was caused by inhalation of lipopolysaccharide (5 mg/kg body weight). The kidneys of intact rats had a typical histological structure without specific features. Histological changes in the renal parenchyma of rats in the study groups included compaction of Malpighian bodies, damage and desquamation of epithelial cells of the nephron tubules, and the appearance of signs of disseminated intravascular coagulation. A month after the start of the experiment, both pathological changes in the nephrons and restored or preserved structural components of the kidney are observed, which indicates activation of intracellular reparative processes. The expression of TGF-β1 fibrosis marker as well as CD68 panmacrophage marker increased on days 3 and 7 of the experiment. The number of macrophages in the kidney samples remained consistently high until the end of the experiment, while the level of TGF-β1 decreased on day 28, indicating the start of the resolution phase. Biochemical analysis of renal markers showed an undulating course of inflammatory processes in the kidneys of experimental rats. The maximum concentration of creatinine, urea, and uric acid in the blood serum was observed at 24 hours of the experiment, which indicated the onset of acute kidney injury as a complication of respiratory distress. Preclinical examination of morphofunctional changes in the kidneys during acute respiratory distress syndrome will help choose an effective method for treating this pathological condition in humans in the future

Investigations on methyl pentanoate-air mixtures confined explosion and cellularity

Combustible fuels like methyl pentanoate (MPe) could cause accidental explosions and fire. Therefore, MPe explosion characteristics have to be well characterized and understood for its safe and reliable use in industries and combustion devices. Considering this, MPe’s explosion pressure, maximum explosion pressure, rate of pressure rise, maximum rate of pressure rise, heat release rate, etc., were investigated in this study at the initial conditions of 1, 2, and 4 bar and 423 K, 453 K, and 483 K using the equivalence ratios of 0.7–1.4. The maximum explosion pressures and maximum rate of pressure rise of MPe increased as the initial pressures increased but the maximum rate of pressure rise was insignificantly affected by the initial temperature. The explosion duration decreased with a decrease in the initial pressure. The maximum heat release rate and deflagration index increased with increasing initial pressure. The MPe maximum deflagration index was 224.8 bar•m/s, which occurred at a higher initial pressure and fuel-rich mixture. Therefore, the explosion severity of MPe was enhanced at higher initial pressures and fuel-rich mixtures. Moreover, the flame cellularity study showed that MPe unstable flame crack length and cell number were more affected by increments in the initial pressure and equivalence ratio. Therefore, flame instability and cellularity would significantly influence explosion severity at higher pressures and equivalence ratios. This study’s findings provide first-hand data that could be used to further investigate and characterize MPe’s explosion risk.

Molecular characteristics of circulating B cells and kidney cells at the single-cell level in special types of primary membranous nephropathy.

Although primary membranous nephropathy (pMN) associated with podocyte autoantibodies (POS) is becoming well-known, the molecular characteristics of the specific type of pMN that is negative for podocyte autoantibodies (NEG) is still unclear. We performed single-cell transcriptome sequencing and single-cell B cell receptor sequencing on circulating CD19+ cells and kidney cells of a NEG paediatric patient with pMN. The single-cell datasets of POS patients and healthy control individuals were included for integrative analysis. The gene expression characteristics and clonal expansion of naïve and memory B cells in the NEG patient changed significantly. We found that a group of CD38+ naïve B cells expanded in the NEG patient, which had the functional characteristics of cell activation. In addition, the conversion between immunoglobulin M (IgM)/IgD and IgG1 in the NEG patient was increased. Parietal epithelial cells (PECs) and podocytes shared similar signature genes (WT1, CLIC5), and new candidate marker genes for PECs, such as NID2, CAV1 and THY1, might contribute to the definition of cell subsets. PECs might have undergone significant changes in the disease, mainly manifested by changes in the expression of CCN2, PLAAT4 and SEPTIN2. The scores of gene sets related to extracellular matrix, cell adhesion and calcium channel in podocytes of the NEG patient was significantly increased. The gene expression of sodium transporter in a group of proximal tubule cells in the disease was significantly increased, especially SLC5A12, which might be related to the oedema of patients. Our research demonstrated the cell type-specific molecular features in the circulation and kidney of the NEG pMN patient.

Delivery and Transcriptome Assessment of an In Vitro Three-Dimensional Proximal Tubule Model Established by Human Kidney 2 Cells in Clinical Gelatin Sponges.

The high prevalence of kidney diseases and the low identification rate of drug nephrotoxicity in preclinical studies reinforce the need for representative yet feasible renal models. Although in vitro cell-based models utilizing renal proximal tubules are widely used for kidney research, many proximal tubule cell (PTC) lines have been indicated to be less sensitive to nephrotoxins, mainly due to altered expression of transporters under a two-dimensional culture (2D) environment. Here, we selected HK-2 cells to establish a simplified three-dimensional (3D) model using gelatin sponges as scaffolds. In addition to cell viability and morphology, we conducted a comprehensive transcriptome comparison and correlation analysis of 2D and 3D cultured HK-2 cells to native human PTCs. Our 3D model displayed stable and long-term growth with a tubule-like morphology and demonstrated a more comparable gene expression profile to native human PTCs compared to the 2D model. Many missing or low expressions of major genes involved in PTC transport and metabolic processes were restored, which is crucial for successful nephrotoxicity prediction. Consequently, we established a cost-effective yet more representative model for in vivo PTC studies and presented a comprehensive transcriptome analysis for the systematic characterization of PTC lines.

Renal Organic Anion Transporters 1 and 3 In Vitro: Gone but Not Forgotten.

Organic anion transporters 1 and 3 (OAT1 and OAT3) play a crucial role in kidney function by regulating the secretion of multiple renally cleared small molecules and toxic metabolic by-products. Assessing the activity of these transporters is essential for drug development purposes as they can significantly impact drug disposition and safety. OAT1 and OAT3 are amongst the most abundant drug transporters expressed in human renal proximal tubules. However, their expression is lost when cells are isolated and cultured in vitro, which is a persistent issue across all human and animal renal proximal tubule cell models, including primary cells and cell lines. Although it is well known that the overall expression of drug transporters is affected in vitro, the underlying reasons for the loss of OAT1 and OAT3 are still not fully understood. Nonetheless, research into the regulatory mechanisms of these transporters has provided insights into the molecular pathways underlying their expression and activity. In this review, we explore the regulatory mechanisms that govern the expression and activity of OAT1 and OAT3 and investigate the physiological changes that proximal tubule cells undergo and that potentially result in the loss of these transporters. A better understanding of the regulation of these transporters could aid in the development of strategies, such as introducing microfluidic conditions or epigenetic modification inhibitors, to improve their expression and activity in vitro and to create more physiologically relevant models. Consequently, this will enable more accurate assessment for drug development and safety applications.