Transcriptional regulatory control of mammalian nephron progenitors revealed by multi-factor cistromic analysis and genetic studies.

Nephron progenitor number determines nephron endowment; a reduced nephron count is linked to the onset of kidney disease. Several transcriptional regulators including Six2, Wt1, Osr1, Sall1, Eya1, Pax2, and Hox11 paralogues are required for specification and/or maintenance of nephron progenitors. However, little is known about the regulatory intersection of these players. Here, we have mapped nephron progenitor-specific transcriptional networks of Six2, Hoxd11, Osr1, and Wt1. We identified 373 multi-factor associated 'regulatory hotspots' around genes closely associated with progenitor programs. To examine their functional significance, we deleted 'hotspot' enhancer elements for Six2 and Wnt4. Removal of the distal enhancer for Six2 leads to a ~40% reduction in Six2 expression. When combined with a Six2 null allele, progeny display a premature depletion of nephron progenitors. Loss of the Wnt4 enhancer led to a significant reduction of Wnt4 expression in renal vesicles and a mildly hypoplastic kidney, a phenotype also enhanced in combination with a Wnt4 null mutation. To explore the regulatory landscape that supports proper target gene expression, we performed CTCF ChIP-seq to identify insulator-boundary regions. One such putative boundary lies between the Six2 and Six3 loci. Evidence for the functional significance of this boundary was obtained by deep sequencing of the radiation-induced Brachyrrhine (Br) mutant allele. We identified an inversion of the Six2/Six3 locus around the CTCF-bound boundary, removing Six2 from its distal enhancer regulation, but placed next to Six3 enhancer elements which support ectopic Six2 expression in the lens where Six3 is normally expressed. Six3 is now predicted to fall under control of the Six2 distal enhancer. Consistent with this view, we observed ectopic Six3 in nephron progenitors. 4C-seq supports the model for Six2 distal enhancer interactions in wild-type and Br/+ mouse kidneys. Together, these data expand our view of the regulatory genome and regulatory landscape underpinning mammalian nephrogenesis.

The Morphology of the Infraorbital Nerve and Foramen in the Presence of an Accessory Infraorbital Foramen.

BACKGROUND: The accessory infraorbital foramen (AIOF) is an anatomical variation associated with the infraorbital foramen (IOF) and nerve (ION). Its occurrence and neural contents have clinical implications regarding failure of loco-regional anesthesia and risk of neural damage during surgical interventions involving the maxillary region. Thus, morphologic characterization of the AIOF and neural contents as well as the spatial relationships to the IOF are potentially useful for optimizing surgical procedures. Additionally, predictive features of the AIOF based on its relationship to IOF morphology could enable the surgeon to anticipate its presence and proceed accordingly. The purpose of this study was to determine whether the presence of an AIOF and its neural contents affected the size, shape, and composition of the IOF and ION. The specific hypothesis tested was that the topography and fascicular composition of the ION and IOF differs between individuals possessing an AIOF and those lacking this anatomical variant. METHODS: Gross topographic features of the IOF (42 crania) were compared between specimens possessing (test) or lacking (control) an AIOF. Nerve fascicles of ION (60 cadaveric sides) were examined histologically and compared morphometrically between specimens presenting or lacking an AIOF. An additional sample of 30 crania was subjected to cone-beam computed tomography (CBCT) analysis to determine the course of the canal leading to the AIOF. RESULTS: The AIOF incidence was 47.6% (20 crania) and 32.1% of the sides (27 sides). A single AIOF was observed in 24 sides and double AIOF in three sides. The AIOF occurred bilaterally in 7 specimens (16.7%). The majority of AIOF (86.7%) were located superomedial to IOF. A slightly higher frequency of the AIOF was found in left side compared to the right. Using CBCT, a patient sample showed an AIOF incidence in 21 sides of 16 patients (65.6%). A single AIOF was observed in 19 sides. Only 1 double AIOF was found in the scans, whereas 3 were found in the dry skull group. The AIOF occurred bilaterally in 3 scans (10%). The majority of AIOF (90.4%) were located superomedial to the IOF based on the CBCT scans. The AIOF was consistently seen connected to the infraorbital canal and progressed superiorly and medially from the infraorbital canal to the maxillary surface. The size of the ION without an AIOF was not significantly different than the ION in the presence of an AIOF (1.45 × 10/1.32 × 10 µm, P < 0.35) based on fascicular area. However, the number of ION fascicles was greater in specimens without an AIOF compared to those showing this feature (15.15/12.71, P < 0.04) CONCLUSION:: Results indicate that the area of the ION is not affected by an AIOF, suggesting that the field of innervation of this area is not modified by its occurrence. However, the ION appears to divide more proximally into its component branches when the AIOF is present.

The society for craniofacial genetics and developmental biology 39th annual meeting.

The Society for Craniofacial Genetics and Developmental Biology (SCGDB) aims to promote education, research, and communication, about normal and abnormal development of the tissues and organs of the head. Membership of the SCGDB is broad and diverse-including clinicians, orthodontists, scientists, and academics-but with all members sharing an interest in craniofacial biology. Each year, the SCGDB hosts a meeting where members can share their latest research, exchange ideas and resources, and build on or establish new collaborations. © 2017 Wiley Periodicals, Inc.

The society for craniofacial genetics and developmental biology 38th annual meeting.

The mission of the Society for Craniofacial Genetics and Developmental Biology (SCGDB) is to promote education, research, and communication about normal and abnormal development of the tissues and organs of the head. The SCGDB welcomes as members undergraduate students, graduate students, post doctoral researchers, clinicians, orthodontists, scientists, and academicians who share an interest in craniofacial biology. Each year our members come together to share their novel findings, build upon, and challenge current knowledge of craniofacial biology. © 2016 Wiley Periodicals, Inc.

Cone beam computed tomography of plastinated hearts for instruction of radiological anatomy.

Radiological anatomy education is an important aspect of the medical curriculum. The purpose of this study was to establish and demonstrate the use of plastinated anatomical specimens, specifically human hearts, for use in radiological anatomy education. Four human hearts were processed with routine plastination procedures at room temperature. Specimens were subjected to cone beam computed tomography and a graphics program (ER3D) was applied to generate 3D cardiac models. A comparison was conducted between plastinated hearts and their corresponding computer models based on a list of morphological cardiac features commonly studied in the gross anatomy laboratory. Results showed significant correspondence between plastinations and CBCT-generated 3D models (98 %; p < .01) for external structures and 100 % for internal cardiac features, while 85 % correspondence was achieved between plastinations and 2D CBCT slices. Complete correspondence (100 %) was achieved between key observations on the plastinations and internal radiological findings typically required of medical student. All pathologic features seen on the plastinated hearts were also visualized internally with the CBCT-generated models and 2D slices. These results suggest that CBCT-derived slices and models can be successfully generated from plastinated material and provide accurate representations for radiological anatomy education.

Midline craniofacial malformations with a lipomatous cephalocele are associated with insufficient closure of the neural tube in the tuft mouse.

BACKGROUND: Genetic variations affecting neural tube closure along the head result in malformations to the face and brain, posing a significant impact on health care costs and the quality of life. METHODS: We have established a mouse line from a mutation that arose spontaneously in our wild-type colony that we called tuft. Tuft mice have heritable midline craniofacial defects featuring an anterior lipomatous cephalocele. RESULTS: Whole-mount skeletal stains indicated that affected newborns had a broader interfrontal suture where the cephalocele emerged between the frontal bones. Mice with a cephalocele positioned near the rostrum also presented craniofacial malformations such as ocular hypertelorism and midfacial cleft of the nose. Gross and histological examination revealed that the lipomatous cephalocele originated as a fluid filled cyst no earlier than E14.5 while embryos with a midfacial cleft was evident during craniofacial development at E11.5. Histological sections of embryos with a midfacial cleft revealed the cephalic neuroectoderm remained proximal or fused to the frontonasal ectoderm about the closure site of the anterior neuropore, indicating a defect to neural tube closure. We found the neural folds along the rostrum of E9 to E10.5 embryos curled inward and failed to close as well as embryos with exencephaly and anencephaly at later stages. Whole-mount in situ hybridization of anterior markers Fgf8 and Sonic hedgehog indicated closure of the rostral site was compromised in severe cases. CONCLUSION: We present a model demonstrating how anterior cranial cephaloceles are generated following a defect to neural tube closure and relevance to subsequent craniofacial morphogenesis in the tuft mouse.

Accessory tympanic plate ossicle: a new osteological entity.

The auricular cartilage, which is typically soft and flexible, can calcify or ossify because of diseases such as diabetes mellitus, trauma, radiation therapy for cancer, and more commonly from frostbite. Calcified, ossified, or hardened auricular cartilage is a rare finding in the clinical literature and appears to be absent in the physical and forensic anthropological literature. This study examines the ossified auricular cartilage and tests whether the hypothesis can be identified in postmortem skeletonized tissue and be part of the external auditory meatus. A total of 290 crania were examined for accessory ossicles. A descriptive and interpretative analysis was performed grossly, histologically, and morphometrically to document the morphology and location of the ossicles, investigate their structure, and perform hypothesis testing. Results revealed that seven females and one male crania from a total of 290 crania (2.76%) exhibit semi-ossified auricular cartilage attached to the tympanic plate of the temporal bone. The morphology and location of the ossicles at the junction of the auricle and external auditory meatus indicate they are hardened auricular cartilage that was verified with histological observations. Regression analysis indicates that addition of the ossicle to the depth of the auditory tube significantly changes coefficient of determination (R2) with respect to cranial breadth. In conclusion, results indicate that small cartilaginous structures of the external ear may ossify forming accessory tympanic plate ossicles that potentially could be identified in skeletal remains as a new osteological entity. This report highlights the types of information that can be gained using an approach that integrates forensic anthropology, gross anatomy, and histology.

Regression analysis of nasal shape from juvenile to adult ages for forensic facial reconstruction.

The nose is a prominent feature for facial recognition and reconstruction. To investigate the relationship of the nasal shape with the piriform aperture in Korean adults and juveniles, we performed regression analysis. By regression analysis, prediction equations for nasal shape were obtained in relation to the shape of the piriform aperture considering sex and age groups. Three-dimensional skull and face models, rendered from computed tomography images, were assessed (331 males and 334 females). Juveniles (<20 years) were divided into three age groups according to the development of the dentition. Adults were divided into three age groups of two decades each, according to their age. To measure the nasal area, nine landmarks and nine measurements were chosen, while seven landmarks and five measurements were selected to measure the piriform aperture area. Four measurements were defined to explain the direct relationship between the nasal aperture and nasal shape. First, descriptive statistical analyses were performed according to sex and age groups. Subsequently, the correlation of nasal soft tissue measurements with piriform measurements was analyzed. Last, we performed a linear regression analysis of the measurements with higher correlations, considering sex and age groups as variables. Prediction equations were used to estimate the nasal bridge length, height, protrusion, and width. Equations considering sex and age groups showed better explanation ability. Measurements related to the height of the nasal bridge presented improvement. This study may assist in the more accurate approximation of nasal shape in facial reconstruction.

A comparison of pre- and post-clinical education learning preferences among medical students who elected to dissect compared to those who did not during the COVID-19 pandemic.

Anatomy instructional methods varied widely during the COVID-19 pandemic and programs are assessing innovations for retention. Learning preferences were assessed among medical students dichotomized as elective dissectors (ED) or non-dissectors (ND) during the COVID-19 partial re-opening in 2020 (preclinical) and again in 2022 after clinical exposure (post-clinical) to assess the viability of elective dissection post-pandemic. A mixed-method approach was used for the assessment of test scores, learning preference surveys, learning activities rankings, and thematic analyses. No significant differences occurred in anatomy examination scores. Dissection was considered useful by both preclinical groups but significantly more so by ED, while the presence of an instructor was significantly preferred by ED although a majority of ND agreed. Elective dissection was significantly preferred by ND but also by a large minority of ED students. Pre- and post-clinical ND believed that elective dissection offered more academic flexibility, did not hinder clinical learning, and did not negatively impact medical education. The corresponding ED stated that confidence improved, clinical experiences were enhanced, and dissection was irreplaceable. Preclinical ND preferred self-learning, while ED students preferred online learning, but these differences largely disappeared post-clinically. Learning activity rankings were not significantly different among all groups (ND, ED, preclinical, and post-clinical). A hybrid laboratory with a virtual learning environment ranked highest across groups and preferences increased over time suggesting that students benefited from this instructional method during clinical exposure. The absence of laboratory experience ranked lowest, and preference decreased over time suggesting that anatomy dissection is valued.

Case of cephalothoracopagus conjoined twinning in an embryonic mouse.

BACKGROUND: Cephalothoracopagus twinning is extremely rare, and it is characterized by fusion of the head and thorax, two separate spines, pelves, and fore- and hindlimbs. CASE: In this case study, we describe cephalothoracopagus twinning in an embryonic mouse displaying a large but exencephalic head, median facial cleft, a single eye, and a second hindbrain rotated roughly 90° from a second spinal cord. There is a bony connection joining the clavicles, resulting in merged asternal thoracic cavities containing two hearts and four lungs. The abdominal cavities contain double caudal digestive tract structures, but a single esophagus and stomach. CONCLUSION: There are several proposed theories regarding the mechanism of spontaneous conjoined twinning; however, the specific mechanisms are still largely unknown. In this report, we highlight the morphological features in a murine example of cephalothoracopagus twinning, furthering our understanding of this rare occurrence while also demonstrating developmental morphogenesis consistent with that reported for human conjoined twins.

Assessment of bone channels other than the nasopalatine canal in the anterior maxilla using limited cone beam computed tomography.

PURPOSE: The anterior maxilla, sometimes also called premaxilla, is an area frequently requiring surgical interventions. The objective of this observational study was to identify and assess accessory bone channels other than the nasopalatine canal in the anterior maxilla using limited cone beam computed tomography (CBCT). METHODS: A total of 176 cases fulfilled the inclusion criteria comprising region of interest, quality of CBCT image, and absence of pathologic lesions or retained teeth. Any bone canal with a minimum diameter of 1.00 mm other than the nasopalatine canal was analyzed regarding size, location, and course, as well as patient gender and age. RESULTS: A total of 67 accessory canals ≥1.00 mm were found in 49 patients (27.8%). A higher frequency of accessory canals was observed in males (33.0%) than in females (22.7%) (p = 0.130). Accessory canals occurred more frequently in older rather than younger patients (p = 0.115). The mean diameter of accessory canals was 1.31 ± 0.26 mm (range 1.01-2.13 mm). Gender and age did not significantly influence the diameter. Accessory canals were found palatal to all anterior teeth, but most frequently palatal to the central incisors. In 56.7%, the accessory canals curved superolaterally and communicated with the ipsilateral alveolar extension of the canalis sinuosus. CONCLUSIONS: The study confirms the presence of bone channels within the anterior maxilla other than the nasopalatine canal. More than half of these accessory bone canals communicated with the canalis sinuosus. From a clinical perspective, studies are needed to determine the content of these accessory canals.

Cholesterol Granuloma in the Maxillary Sinus: A Rare Presentation Associated With an Odontogenic Cyst.

Cholesterol granuloma is a histopathological finding characterized by a mass of connective tissue and granulation tissue. It is primarily observed in the middle ear, mastoid process, or paranasal sinuses, with rare occurrences in the dental odontogenic region. A dentigerous cyst encloses the crown of an unerupted tooth by expanding its follicle and attaches to the neck of the tooth. Here, we report a 63-year-old female who presented to the dental clinic complaining of an ill-fitted denture. A panoramic radiograph showed a well-defined radiolucent lesion in the upper left maxillary sinus with an impacted third molar. Computed tomography revealed loss of the anterior and lateral sinus walls. The cyst was enucleated surgically. The final diagnosis was confirmed by histopathological examination, which revealed focal areas of cholesterol clefts in the cystic wall of the dentigerous cyst.

Themes in neuronavigation research: A machine learning topic analysis.

Objective: To understand trends in neuronavigation we employed machine learning methods to perform a broad literature review which would be impractical by manual inspection. Methods: PubMed was queried for articles with "Neuronavigation" in any field from inception-2020. Articles were designated neuronavigation-focused (NF) if "Neuronavigation" was a major MeSH. The latent dirichlet allocation topic modeling technique was used to identify themes of NF research. Results: There were 3896 articles of which 1727 (44%) were designated as NF. Between 1999-2009 and 2010-2020, the number of NF publications experienced 80% growth. Between 2009-2014 and 2015-2020, there was a 0.3% decline. Eleven themes covered 1367 (86%) NF articles. "Resection of Eloquent Lesions" comprised the highest number of articles (243), followed by "Accuracy and Registration" (242), "Patient Outcomes" (156), "Stimulation and Mapping" (126), "Planning and Visualization" (123), "Intraoperative Tools" (104), "Placement of Ventricular Catheters" (86), "Spine Surgery" (85), "New Systems" (80), "Guided Biopsies" (61), and "Surgical Approach" (61). All topics except for "Planning and Visualization", "Intraoperative Tools", and "New Systems" exhibited a monotonic positive trend. When analyzing subcategories, there were a greater number of clinical assessments or usage of existing neuronavigation systems (77%) rather than modification or development of new apparatuses (18%). Conclusion: NF research appears to focus on the clinical assessment of neuronavigation and to a lesser extent on the development of new systems. Although neuronavigation has made significant strides, NF research output appears to have plateaued in the last decade.

Extended reality veterinary medicine case studies for diagnostic veterinary imaging instruction: Assessing student perceptions and examination performance.

Educational technologies in veterinary medicine aim to train veterinarians faster and improve clinical outcomes. COVID-19 pandemic, shifted face-to-face teaching to online, thus, the need to provide effective education remotely was exacerbated. Among recent technology advances for veterinary medical education, extended reality (XR) is a promising teaching tool. This study aimed to develop a case resolution approach for radiographic anatomy studies using XR technology and assess students' achievement of differential diagnostic skills. Learning objectives based on Bloom's taxonomy keywords were used to develop four clinical cases (3 dogs/1 cat) of spinal injuries utilizing CT scans and XR models and presented to 22 third-year veterinary medicine students. Quantitative assessment (ASMT) of 7 questions probing 'memorization', 'understanding and application', 'analysis' and 'evaluation' was given before and after contact with XR technology as well as qualitative feedback via a survey. Mean ASMT scores increased during case resolution (pre 51.6% (±37%)/post 60.1% (± 34%); p < 0.01), but without significant difference between cases (Kruskal-Wallis H = 2.18, NS). Learning objectives were examined for six questions (Q1-Q6) across cases (C1-4): Memorization improved sequentially (Q1, 2 8/8), while Understanding and Application (Q3,4) showed the greatest improvement (26.7%-76.9%). Evaluation and Analysis (Q5,6) was somewhat mixed, improving (5/8), no change (3/8) and declining (1/8).Positive student perceptions suggest that case studies' online delivery was well received stimulating learning in diagnostic imaging and anatomy while developing visual-spatial skills that aid understanding cross-sectional images. Therefore, XR technology could be a useful approach to complement radiological instruction in veterinary medicine.

Predicting the eyebrow from the orbit using three-dimensional CT imaging in the application of forensic facial reconstruction and identification.

Eyebrows are the most important facial feature in facial recognition with its shape rated to be more helpful than color or density for facial reconstruction or approximation. However, little extant research has estimated the position and morphological territory of the eyebrow from the orbit. Three-dimensional craniofacial models, produced from CT scans of 180 Koreans autopsied at the National Forensic Service Seoul Institute, were used to conduct metric analyses of subjects (125 males and 55 females) between 19 and 49 (mean 35.1) years. We employed 18 craniofacial landmarks to examine the morphometry of the eyebrow and orbit with 35 pairs of distances per subject measured between landmark and reference planes. Additionally, we used linear regression analyses to predict eyebrow shape from the orbit for every possible combination of variables. The morphology of the orbit has more influence on the position of the superior margin of the eyebrow. In addition, the middle part of the eyebrow was more predictable. The highest point of the eyebrow in female was located more medially than the male. Based on our findings, the equations for estimating the position of the eyebrow from the shape of the orbit is useful information for face reconstruction or approximation.

Craniofacial features resembling frontonasal dysplasia with a tubulonodular interhemispheric lipoma in the adult 3H1 tuft mouse.

Intracranial lipomas are rare, but 45% of them occur along the midline cisterns between the hemispheres and are often associated with corpus callosum hypoplasia and craniofacial defects. They are difficult to detect as they are generally asymptomatic and visible by MRI or by postmortem examination. The exact cause of these interhemispheric lipomas is not known, but they arise from a developmental defect resulting in the maldifferentiation of mesenchymal cells into mesodermal derivatives that are not normally present. We have identified a new mouse mutant called tuft, exhibiting a forebrain, intracranial lipoma with midline craniofacial defects resembling frontonasal dysplasia (FND) that arose spontaneously in our wild-type 3H1 colony. The tuft trait seems to be transmitted in recessive fashion, but approximately 80% less frequent than the expected Mendelian 25%, due to either incomplete penetrance or prenatal lethality. MRI and histologic analysis revealed that the intracranial lipoma occurred between the hemispheres and often protruded through the sagittal suture. We also observed a lesion at the lamina terminalis (LT) that may indicate improper closure of the anterior neuropore. We have mapped the tuft trait to within an 18 cM region on mouse chromosome 10 by microsatellite linkage analysis and identified several candidate genes involved with craniofacial development and cellular differentiation of adipose tissue. Tuft is the only known mouse model for midline craniofacial defects with an intracranial lipoma. Identifying the gene(s) and mutation(s) causing this early developmental defect will help us understand the pathogenesis of FND and related craniofacial disorders.

Extended reality visualization of medical museum specimens: Online presentation of conjoined twins curated by Dr. Jacob Henle between 1844-1852.

Background: The purpose of this study is to characterize a full-term conjoined twins' cadaver curated by Dr. Jacob Henle sometime between 1844 and 1852 and demonstrate digital distribution of an old and rare medical museum specimen using an extended reality (XR) model workflow. Methods: The cadaver (Preparation 296) is in the Department of Anatomy and Cell Biology at the University of Heidelberg. An XR display workflow comprises image capture, segmentation, and visualization using CT/MR scans derived from the cadaver. Online radiology presentation to medical students focuses on diagnostic characteristics of anatomical systems depicted with XR models. Results: Developmental defects in Preparation 296 include duplicated supradiaphragmatic structures and abnormal osteological features. Subdiaphragmatically, the gut is continuous on the right, but terminates at the distal esophagus on the left. One large liver occupies the abdomen with one spleen located on the left side. Observations suggest duplication of the primitive streak and separate notochords rostrally. Duplication occurs near the yolk sac and involves midgut formation while secondary midline fusion of the upper extremities and ribs likely results from the proximity of the embryos during development. Medical students access the model with device agnostic software during the curricular topic "Human Body Plan" that includes embryology concepts covering mechanisms of twinning. Conclusions: The workflow enables ease-of-access XR visualizations of an old and rare museum specimen. This study also demonstrates digital distribution and utilization of XR models applicable to embryology education.

Case-based radiological anatomy instruction using cadaveric MRI imaging and delivered with extended reality web technology.

PURPOSE: Extended reality (XR) technology enhances learning in medical education. The purpose of this study was to develop and apply a case-based approach for teaching radiological anatomy utilizing XR technology for improved student exploration and engagement. METHODS: The workflow consisted of MRI scanning cadavers followed by radiological, pathological, and anatomical assessment, and finally case presentation based on XR visualizations and student interaction. Case information (Subject, History, and Physical Exam) was presented to student groups who generated and recorded hypotheses using Google Forms. RESULTS: Use of all components of the system was voluntary and a total of 74 students responded to the survey request (response rate = 95%). Assessment of the experience was conducted through a qualitative survey comprising four Likert scale questions (1-5, 1 lowest), three binary questions, and open-ended comments. Mean, standard deviation, and overall agreement (mean ± SD, OA) showed that students found MRI scans of cadavers to be helpful for dissections (4.14 ± 1.1, 74.3%) and provided an understanding of relevant anatomy (4.32 ± 0.9, 79.7%), while 78.4% of students used the DICOM viewer to visualize scans of cadavers. The difficulty of use was found to be average (2.90 ± 1.0, 23%). zSpace visualizations were used by 40.5% of students, generally agreeing that an understanding of spatial relationships improved as a result (3.60 ± 1.0, 43.2%). More case-based sessions were favored by 97.3% of students. CONCLUSIONS: Results suggest that cadaveric MRI radiological visualization and XR technology enhance understanding of case-based anatomical dissections and encourage student exploration and engagement.

Osmoregulatory defect in adult mice associated with deficient prenatal expression of six2.

Suboptimal kidney development resulting from a genetic deficit in nephron number can have lifelong consequences that may lead to cardiorenal complications upon exposure to secondary insults in later life. To determine whether the inherited reduced renal reserve compromises the ability to handle osmotic stress in the adult animal, we challenged the heterozygous 3H1 Brachyrrhine (Br/+) mouse, which displays heritable renal hypoplasia associated with reduced embryonic six2 expression, to a solution of 2% NaCl for 5 days or to fluid restriction for 48 h. Blood chemistry, fluid intake, and physiological parameters, including renal measurements, were determined. Systemic hypertonicity by prolonged salt loading led to significant increases in plasma osmolality and plasma Na(+), along with polydipsia and polyuria, with a significant urine-concentrating defect that was resistant to DDAVP treatment in the adult Br/+ mouse compared with wild-type littermates. The Br/+ mouse also developed a significant increase in blood urea nitrogen at baseline that was further elevated when 2% NaCl was given. Fluid restriction for 48 h further enhanced plasma osmolality and plasma Na(+) responses, although the Br/+ mouse was evidently able to produce a small amount of concentrated urine at this time. Hypothalamic c-Fos expression was appropriately activated in the Br/+ mouse in response to both osmotic challenges, indicating an intact central neuroendocrine pathway that was not affected by the lack of congenital six2 expression. Collectively, our results demonstrate impaired osmoregulatory mechanisms consistent with chronic renal failure in the Br/+ mouse and indicate that six2 haploinsufficiency has a direct effect on postnatal fluid and electrolyte handling associated with fluid imbalance.