Biomedical subjects

L A Opperman

Publications and source records attributed to L A Opperman.

At least 19 recordsLinked to original sources

PubMed2006

Cellular response to force application at craniofacial sutures.

OBJECTIVES: To provide a comprehensive review of the literature describing research done on the responses of suture cells to force application in vitro and in vivo. DESIGN AND RESULTS: This review outlines the types of forces that can be applied, methods of applying the forces, the sutures used in experiments, and the changes in morphology, molecular biology (gene and protein expression), and cell biology (proliferation, differentiation, apoptosis) in response to these forces. CONCLUSION: The molecular response of sutures to force needs to be further investigated as these molecules can be used to enhance the way in which craniofacial sutures respond to mechanical force during orthopedic-orthodontic treatment.

Apoptosis↗

PubMed2003

Osteoblasts and MG-63 osteosarcoma cells behave differently when in contact with ProRoot MTA and White MTA.

AIM: To test the hypothesis that MG-63 osteosarcoma cells and primary osteoblasts react differently to ProRoot trade mark MTA (mineral trioxide aggregate) and White MTA by: (i) investigating the attachment of primary osteoblasts and MG-63 osteosarcoma cells to ProRoot trade mark MTA and White MTA; and (ii) comparing the osteogenic behaviour of both cell lines in contact with these endodontic materials. METHODOLOGY: Primary osteoblasts were harvested from foetal rat calvaria by sequential digestion and MG-63 osteosarcoma cells were purchased. Cells were exposed to ProRoot trade mark MTA and White MTA prepared according to the manufacturer's instructions. All samples and controls were prepared in quadruplicate. After 6, 9 and 13 days exposure to MTA, the cells were fixed and prepared for SEM examination. In addition, both the cell types were grown to confluence and exposed to beta-glycerophosphate and dexamethasone to assess mineralized nodule formation as a function of osteogenic behaviour. RESULTS: The number of cells on the surface of the culture dish and on top of the materials increased in all samples throughout the 3 time periods, except for White MTA where no primary osteoblasts were visible on top of the material by the end of 13 days. After exposing cells to differentiation medium nodules were observed in cultures of primary osteoblasts, but not of MG-63 osteosarcoma cells. CONCLUSIONS: Under the conditions of this study, whilst primary osteoblasts initially bound to White MTA, they did not survive on the surface by the end of 13 days. Primary osteoblasts formed mineralized nodules when exposed to differentiation medium, whilst MG-63 cells did not form nodules. As MG-63 cells do not behave osteogenically by forming mineralized nodules, and primary osteoblasts are more sensitive than MG-63 osteosarcoma cells to White MTA in cell culture, primary osteoblasts are more appropriate than MG-63 cells for testing endodontic materials in cell culture.

Aluminum Compounds↗

PubMed2002

Transforming growth factor-beta3 (Tgf-beta3) down-regulates Tgf-beta3 receptor type I (Tbetar-I) during rescue of cranial sutures from osseous obliteration.

Appropriate biochemical regulation of intramembranous bone growth from sutures is necessary to achieve correct craniofacial morphology. Failure to form sutures (agenesis) or to maintain sutures in their unossified state (craniosynostosis) can result in severe facial dysmorphology. Several factors such as Twist, Msx2, fibroblast growth factors (Fgfs), bone morphogenetic proteins (Bmps) and transforming growth factors-beta (Tgf-betas) regulate suture patency, likely by interacting with one another. Tgf-beta2 and Tgf-beta3 use the same cell surface receptors, yet have opposite effects on suture patency, cellular proliferation and apoptosis within the suture. One possible mechanism by which Tgf-beta3 rescues sutures from obliteration is by regulating the ability of suture cells to respond to Tgf-beta2. As Tgf-beta3 does not regulate protein levels of Tgf-beta2 in sutures, Tgf-beta3 could regulate tissue responsiveness to Tgf-beta2 by regulating Tgf-beta2 access to receptors. Tgf-beta3 is a more potent competitor than Tgf-beta2 for cell surface receptors, so it is proposed that Tgf-beta3 binds to and down-regulates Tgf-beta receptor type I (Tbetar-I) expression by suture cells. This down-regulation would limit the ability of cells to respond to all Tgf-betas, including Tgf-beta2. To test this hypothesis, an in vitro culture model was used in which fetal rat sutures either remain patent or are induced to fuse when cultured in the presence or absence of dura mater, respectively. Tgf-beta3 was added to cultured calvaria and changes in the number of receptor positive cells within the suture were established. Data were compared with that seen in control sutures and in normal sutures in vivo. It was found that the numbers of cells expressing Tbetar-I within the suture matrix increased over time in sutures remaining patent. Osteoblastic cells lining the bone fronts on either side of sutures were Tbetar-I positive during early morphogenesis, but these numbers declined as sutures fused, both in vivo and in vitro. Addition of Tgf-beta3 to calvaria in culture decreased the number of Tbetar-I expressing cells in both fusing and non-fusing sutures, with dramatic decreases in the numbers of osteoblasts expressing Tbetar-I.

Activin Receptors, Type I↗

PubMed2002

Tgf-beta1, Tgf-beta2, Tgf-beta3 and Msx2 expression is elevated during frontonasal suture morphogenesis and during active postnatal facial growth.

OBJECTIVES: It is hypothesized that regulation of facial suture morphogenesis is similar to that of cranial sutures, with expression of similar regulatory molecules, governing suture formation and patency. The present study was designed to characterize the morphology of the frontonasal (FN) suture of the rat at different developmental stages and to investigate the presence and temporal-spatial expression of transforming growth factor-beta 1 (Tgf-beta1), Tgf-beta2, Tgf-beta3 and Msx2 mRNA within these structures. SETTING AND SAMPLE POPULATION: The Department of Biomedical Sciences at Texas A&M University System Health Science Center, Baylor College of Dentistry, Dallas, TX USA. Histological sections and RNA isolated from FN suture tissues of Sprague-Dawley rats, aged embryonic day 16 through postnatal day 20. METHOD: Sections were examined after immunohistochemical staining. Gene expression was determined by densitometric analysis of RT-PCR products run on agarose gels. RESULTS: FN sutures develop slightly later than cranial sutures and show increased complexity over time when compared to cranial sutures. FN sutures were closely associated with the nasal capsular cartilage, with intervening layers of perichondrium and periosteum. The pattern of expression of Tgf-betas within the FN suture tissues was similar to that seen in the cranial sutures. However, mRNA and protein of the Tgf-betas were differentially expressed over time compared to cranial sutures. In FN sutures, Tgf-beta mRNA levels were elevated both during the period of suture morphogenesis and during active bone growth from the suture in the early postnatal period. Msx2 mRNA expression was elevated in both the prenatal and postnatal periods, similar to Tgf-beta mRNA expression. CONCLUSION: Tgf-beta and Msx2 are present in facial sutures similar to cranial sutures, but are differentially expressed over time, perhaps reflecting different bone growth rates from these sutures.

Animals↗

PubMed2001

Effect of recombinant human bone morphogenetic protein-2 on bone regeneration and osseointegration of dental implants.

Recombinant human bone morphogenetic protein-2 (rhBMP-2) induced bone regeneration and osseointegration was evaluated in bony defects created within the hollow chamber of endosseous dental implants in 14 foxhound dogs. Bilateral extractions of mandibular premolars were performed and surgical implantation of 104 hollow cylinder implants followed after 8 weeks of healing. Experimental implants had their hollow chamber filled with 20 microg of rhBMP-2 delivered with a bovine collagen carrier, whereas the control implants had their apical chamber left empty. Dogs were followed for 2, 4, 8 and 12 weeks. Histomorphometric evaluation and immunohistochemical analysis were performed. Minimal bone was regenerated at 2 weeks for both groups. At 4 weeks, bone fill averaged 23.48% for the rhBMP-2 and 5.98% for the control group (P<0.05). At 8 weeks, mean bone fill was 20.94% and 7.75% for the rhBMP-2 and the controls, respectively (P<0.05). At 12 weeks, mean bone fill was 31.39% and 24.31% for the rhBMP-2 and control implants, respectively (P>0.05). Bone-implant contact (BIC) increased for both groups over time and at 8 weeks the rhBMP-2 BIC value was 18.65% and for the control 7.22% (P<0.05). At 12 weeks, the BIC was 43.78% and 21.05% for the rhBMP-2 and the control group, respectively (P<0.05). Immunohistochemical staining for type II collagen was positive only for parts of the collagen carrier and formation of cartilaginous intermediate was not observed in any of the specimens. The results suggest that, in confined defects adjacent to dental implants, rhBMP-2 can induce bone regeneration in close apposition to the implant surface.

Animals↗

PubMed2001

Correction of coronal suture synostosis using suture and dura mater allografts in rabbits with familial craniosynostosis.

OBJECTIVE: Resynostosis following surgical correction of craniosynostosis is a common clinical correlate. Recent studies suggest that the dura mater is necessary to maintain suture patency. It has also been hypothesized that dura mater from synostotic individuals may provide aberrant biochemical signals to the osteogenic fronts of the calvaria, which result in premature suture fusion and subsequent resynostosis following surgery. This study was designed to test this hypothesis by surgically manipulating the coronal suture and dura mater in rabbits with familial craniosynostosis to prevent postsurgical resynostosis. DESIGN: Craniofacial growth and histomorphometric data were collected from 129 rabbits: 72 normal controls and 57 rabbits with bilateral coronal suture synostosis (15 unoperated on controls; 13 surgical controls; 9 dura mater transplant only; 10 suture transplant only; and 10 suture and dura mater transplant). At 10 days of age, all rabbits had radiopaque amalgam markers placed on either side of the coronal, frontonasal, and anterior lambdoidal sutures. At 25 days of age, 42 synostosed rabbits had a 3 to 5-mm wide coronal suturectomy. Coronal sutures and/or underlying dura mater allografts were harvested from same-aged, wild-type, isohistogenic control rabbits and transplanted onto the dura mater of synostosed host rabbits. Serial radiographs were taken at 10, 25, 42, and 84 days of age, and the suturectomy sites were harvested at 84 days of age in 44 rabbits and serially sectioned for histomorphometric examination. RESULTS: Results revealed that cranial vault growth was significantly (p < .05) improved following surgical release of the fused coronal suture compared with synostosed rabbits who were not operated on but was still significantly different (p < .05) from that of normal control rabbits. By 84 days of age, significant (p < .05) differences were noted in calvarial suture marker separation, cranial vault shape indices, and cranial base angles between rabbits with and without dura mater allografts, probably as a result of resynostosis of the suturectomy site or suture-only allografts. Qualitative histological examination revealed that at 84 days of age rabbits with suture and dura allografts had patent coronal sutures, suture-only allografts had fused coronal sutures with extensive endosteal hyperostosis, dura mater-only allografts had some new bone in the suturectomy site that resembled rudimentary osteogenic fronts, and suturectomy controls had extensive endosteal bone formation and resynostosis of the suturectomy site. Significantly (p < .05) more bone was found in the suturectomy sites of rabbits without dura mater allografts compared with rabbits with dura mater allografts. CONCLUSIONS: Results support the initial hypothesis that normal dura mater allografts will maintain suture or suturectomy site patency and allow unrestricted craniofacial growth. However, it is still unclear whether the dura mater from normal rabbits was providing biochemical signals to the transplanted sutures or suturectomy sites or simply acting as a barrier to prevent abnormal biochemical signals from the dura mater of synostosed rabbits from reaching the calvaria. The clinical and therapeutic implications of these procedures are discussed.

Analysis of Variance↗

PubMed2000

Transforming growth factor-beta 2 and TGF-beta 3 regulate fetal rat cranial suture morphogenesis by regulating rates of cell proliferation and apoptosis.

Cranial vault sutures are the major intramembranous bone growth sites during rapid expansion of the neurocranium. To function as bone growth sites, sutures need to remain patent, while allowing rapid bone formation at the edges of the bone fronts. Premature osseous obliteration of sutures (craniosynostosis) by fusion of bone fronts across the suture site prevents further bone formation at this site, often leading to severe facial dysmorphology. Although several growth factor receptor and transcription factor mutations have been implicated in craniosynostosis, the underlying mechanisms leading to sutural obliteration remain unclear. Previous studies have shown that dura secreted soluble factors responsible for maintaining suture patency and that suture fusion observed in the absence of dura was preceded by elevated levels of DNA synthesis and collagen production in the suture region. The use of neutralizing antibodies in a fetal calvarial culture model further demonstrated that removal of transforming growth factor (TGF) -beta 3 activity induced premature sutural obliteration, whereas removal of TGF-beta 2 activity prevented sutural obliteration. Data presented here demonstrate that suture obliteration induced by removal of TGF-beta 3 activity was preceded by elevated levels of DNA synthesis, similar to that seen upon removal of the dura. Addition of exogenous TGF-beta 3 to calvaria cultured without dura both prevented suture obliteration and reduced DNA synthesis to levels comparable to those seen with intact dura. Addition of exogenous TGF-beta 2 to calvarial cultures induced sutural fusion accompanied by elevated levels of cell proliferation. However, sutures rescued from obliteration by removal of TGF-beta 2 activity did not have decreased levels of cell proliferation, but rather appeared to be due to inhibited differentiation. In all calvaria in which sutures remained patent in culture, numbers of apoptotic cells were high within the suture, whereas in sutures destined to fuse, numbers of apoptotic cells were low. Results indicate that one of the critical regulators of suture patency is cell number. Alterations in cell number can trigger premature differentiation of cells, resulting in sutural obliteration. Furthermore, a complex interplay between closely related molecules is required to maintain cranial vault sutures in an unossified state, while allowing new bone to be formed at the edges of the bone fronts.

Animals↗

PubMed2000

Cranial sutures as intramembranous bone growth sites.

Intramembranous bone growth is achieved through bone formation within a periosteum or by bone formation at sutures. Sutures are formed during embryonic development at the sites of approximation of the membranous bones of the craniofacial skeleton. They serve as the major sites of bone expansion during postnatal craniofacial growth. For sutures to function as intramembranous bone growth sites, they need to remain in an unossified state, yet allow new bone to be formed at the edges of the overlapping bone fronts. This process relies on the production of sufficient new bone cells to be recruited into the bone fronts, while ensuring that the cells within the suture remain undifferentiated. Unlike endochondral growth plates, which expand through chondrocyte hypertrophy, sutures do not have intrinsic growth potential. Rather, they produce new bone at the sutural edges of the bone fronts in response to external stimuli, such as signals arising from the expanding neurocranium. This process allows growth of the cranial vault to be coordinated with growth of the neurocranium. Too little or delayed bone growth will result in wide-open fontanels and suture agenesis, whereas too much or accelerated bone growth will result in osseous obliteration of the sutures or craniosynostosis. Craniosynostosis in humans, suture fusion in animals, and induced suture obliteration in vitro has been associated with mutations or alterations in expression of several transcription factors, growth factors, and their receptors. Much of the data concerning signaling within sutures has been garnered from research on cranial sutures; hence, only the cranial sutures will be discussed in detail in this review. This review synthesizes classic descriptions of suture growth and pathology with modern molecular analysis of genetics and cell function in normal and abnormal suture morphogenesis and growth in a unifying hypothesis. At the same time, the reader is reminded of the importance of the suture as an intramembranous bone growth site.

Animals↗

PubMed2000

A pilot study of short- and long-term sequelae to rigid fixation across metacarpal physes in a baboon model.

The use of rigid fixation for fractures of the extremities has become commonplace. The short- and long-term effects of rigid fixation on the growing hand, however, have not been studied thoroughly. In this project, the use of rigid fixation across metacarpal growth plates (physes) in growing primate hands was examined. The hypothesis to be tested was that long-term placement of rigid fixation devices across physes during stabilization of mid-shaft osteotomies will cause the physes to close prematurely. Fixation devices with screws placed in the epiphysis and left in place for 4 months or 1 year resulted in open physes, in support of the null hypothesis. However, in physes plated for 1 year, biochemical changes associated with increased bone differentiation were apparent. Findings suggest that rigid fixation across physes for as long as 1 year can be used appropriately in growing individuals when necessary. However, until additional investigation establishes whether the open physes are still capable of producing bone-lengthening hypertrophic chondrocytes, caution should be used in long-term placement of rigid fixation devices.

Animals↗

PubMed1997

TGF-beta 1, TGF-beta 2, and TGF-beta 3 exhibit distinct patterns of expression during cranial suture formation and obliteration in vivo and in vitro.

Cranial sutures function as bone growth centers while themselves remaining unossified. Rat frontonasal sutures become obliterated by neonatal day 21 (N21), while coronal sutures do not fuse over the life of the animal. Coronal sutures induced to undergo osseous obliteration in vitro after removal of the dura mater were found to require soluble, heparin-binding factors present in dura mater to resist osseous obliteration. Transforming growth factor beta 1 (TGF-beta 1), beta 2, and beta 3, heparin-binding factors known to regulate bone cell proliferation and differentiation, were considered likely candidates. The presence and distribution of these factors in calvarial tissues both in vivo and in vitro were established by immunohistochemical analysis, while reverse transcription followed by polymerase chain reaction (RT/PCR) was employed to determine the presence of transcripts for these factors in mRNA isolated from microdissected dura mater. Results indicated that the presence of TGF-beta 1 and TGF-beta 2 were associated with developing coronal and frontonasal sutures, and that the continued presence of these factors was associated with osseous obliteration of the frontonasal suture. However, increased TGF-beta 3 immunoreactivity was associated with the coronal suture remaining unossified. RT/PCR demonstrated the presence of transcripts for TGF-beta 1, beta 2, and beta 3 in dural tissues isolated from rat calvaria. These data support the notion of a role for TGF-beta s in regulating cranial suture morphogenesis and establish the in vitro model as a valid system for examining mechanisms by which growth factors regulate both suture morphogenesis and bone growth at the suture site.

Animals↗

PubMed1997

Elevated levels of transforming growth factors beta 2 and beta 3 in lambdoid sutures from children with persistent plagiocephaly.

OBJECTIVE: To analyze the pertinent history and physical findings specific to the subset of patients with a progressive posterior skull deformity, requiring surgery to correct their deformity. PATIENTS: Since the Academy of Pediatrics issued its recommendation on supine positioning of infants to prevent sudden infant death syndrome (SIDS) in 1992, 73 children have presented to the University of Virginia Craniofacial Anomalies Clinic with posterior-skull deformities. The majority were successfully managed with conservative therapy, but in six patients, the deformity was severe and persistent, requiring surgical correction. All six children were older (7.5-12 mo), presenting with more severe morphologic appearances and a higher incidence of associated neurodevelopmental delay. Three had family backgrounds of isolated craniosynostosis. METHODS: Characteristics of these patients were examined to determine why they may have differed from those that responded to conservative management. Immunohistochemical staining of their lambdoid sutures was performed. RESULTS: Significantly increased staining for TGF-beta 2 and TGF-beta 3, potent stimulators of bone cell growth and differentiation, was seen in all 'affected' sutures from the flattened side of the skull, compared to unaffected sutures from the protruding side of the skull-a pattern similar to that seen during normal bony obliteration of calvarial sutures. CONCLUSION: The majority of patients with posterior plagiocephaly associated with positioning responded to conservative management, while a small subset of patients with persistent posterior skull deformation required surgical intervention. A genetic basis for the latter patients' persistent plagiocephaly, rather than positioning, cannot be ruled out. Genetics, prolonged external pressure against the sutures, or a combination of these factors may lead to permanently raised levels of growth factors in 'affected' sutures.

Cell Differentiation↗

PubMed1995

Cranial sutures require tissue interactions with dura mater to resist osseous obliteration in vitro.

A chemically defined serum-free medium, which supports the development of bones and fibrous tissues of rat calvaria from nonmineralized mesenchymal precursor tissues, was employed to investigate tissue interactions between the dura matter and overlying tissues. Fetal calvarial rudiments from stages prior to bone and suture morphogenesis (fetal days 19 and 20) and neonatal calvarial rudiments with formed sutures (day 1) were cultured with and without associated dura mater. Removal of calvaria for in vitro culture allowed the examination of suture morphogenesis in the absence of tensional forces exerted on the sutures via fiber tracts in the dura mater originating in the cranial base. Ossification of frontal and parietal bones proceeded in a fashion comparable to development in vivo, but the cranial (coronal) sutures--primary sites for subsequent skull growth--were obliterated by osseous tissue union in the absence of dura mater. Bony fusion did not occur when rudiments were cocultured with dura mater on the opposite sides of 0.45 microns polycarbonate transwell filters, suggesting that the influence of dura mater on sutural obliteration was mediated by soluble factors rather than cell-cell or cell-matrix interactions. These results indicate that cell signaling mechanisms rather than biomechanical tensional forces are required for morphogenesis of the calvaria.

Animals↗

PubMed1995

Repair of critical size rat calvarial defects using extracellular matrix protein gels.

In this study the authors examined the capacity of gels of reconstituted basement membrane, laminin, and type I collagen to mediate repair of critical size defects in rat calvaria. Although autografts are widely used to repair bone defects caused by trauma or surgical treatment of congenital malformations, neoplasms, and infections, an adequate quantity of graft is not always available. Allogenic bone is readily available, but its use is associated with an increased incidence of nonunion, fatigue fracture, and rejection. Biologically active, purified components of basement membranes, which have been shown to promote osteogenic differentiation and angiogenesis in vitro and type I collagen (the major constituent of bone extracellular matrix) can be formed into native isotonic space-filling gels. In this study critical size calvarial defects were created in retired male Sprague-Dawley rats. Thirty-six animals were divided into seven groups. Group 1 (control) received no treatment for the defects. Group 2 animals were implanted with methylcellulose. Groups 3, 4, 5, and 6 were implanted with gels of type I collagen, reconstituted basement membrane, or laminin, respectively. The last group of three animals (Group 7) was implanted with 100 micrograms of type I collagen gels (identical to Group 3) and sacrificed at 20 weeks following a single CT scan to determine if complete healing could be obtained with this method given sufficient time. Except for rats in the type I collagen group that was evaluated by multiple computerized tomography (CT) scans biweekly from 2 to 12 weeks, bone repair was evaluated using CT at 12 weeks. Healing was quantified using three-dimensional reconstruction of CT. Following the final CT scan in each experimental group, animals were sacrificed, and a sample of tissues was evaluated by conventional histology. Animals treated with type I collagen gels showed 87.5% repair of the area of the defects at 12 weeks and 92.5% repair by 20 weeks. Increasing the gel volume 1.5 x accelerated complete repair to 3 months. Murine-reconstituted basement membrane and laminin gels induced 55.5% and 46.3% repair, respectively, at 3 months. In untreated control animals 7% repair of the area of the defects showed at 3 months. Histological analysis confirmed new bone formation in partial and completely healed defects. Bioengineered native collagen gels may have wide applicability for bone repair as an alternative bone graft material alone, in combination with autograft or marrow aspirate, or as a delivery system for osteogenic growth factors.

Animals↗

PubMed1994

In the absence of periosteum, transplanted fetal and neonatal rat coronal sutures resist osseous obliteration.

Normal craniofacial development depends on expansion of the cranial vault by growth at the sutures. Inappropriate development of the sutures leads to global disruption of patterns of craniofacial growth. Tissue interactions between dura mater and suture matrix play a critical role in the phenotypic maintenance of cranial sutures. However, the function of the periosteum in this process remains under-reported and controversial. To examine the contribution of periosteum in maintaining the patency of coronal sutures, fetal and neonatal rat coronal sutures were transplanted to surgically created defects in adult rat host parietal bones. These sutures were examined for their ability to persist in the host milieu in the presence and absence of both donor and host periosteum. This study established that removal of both host and transplant periosteum, unlike removal of dura mater, did not lead to obliteration of either fetal or neonatal sutures. Thus, periosteum and dura mater are nonequivalent tissues with respect to influence on suture patency.

Animals↗

PubMed1994

Subterranean mole-rats naturally have an impoverished calciol status, yet synthesize calciol metabolites and calbindins.

Mole-rats (Family Bathyergidae) have no obvious source of calciol. They live in an environment devoid of sunlight and consume a herbivorous diet. Calciol status, metabolism and expression were examined in six species of Bathyergids. Serum levels of calcidiol in all species were < 5 micrograms/l and those of calcitriol were low (18.0 +/- 11.0 (SD) ng/l, N = 57) when compared to other rodents. Within 72 h of injecting animals with tritium-labelled calciol, most of the labelled prohormone had been metabolized to more polar metabolites. Three times more tritium-labelled calcitriol (19.3 +/- 2.9%) was present than (24R)-hydroxycalcidiol (6.2 +/- 10%). The natural absence of detectable circulating concentrations of calcidiol and the threefold greater amount of calcitriol to (24R)-hydroxycalcidiol produced indicate that calciol naturally is in short supply. Calciol-dependent calbindins were absent in the duodenum. Calbindin-D28k was present in the Purkinje cells of the cerebellum and in some collecting ducts and proximal and distal convoluted tubules of the kidney. Calbindin-D9k also was present but was localized uniquely in the juxtaglomerular cells of the five southern African species. These data confirm that Bathyergid mole-rats naturally have an impoverished calciol status. Despite the presence of calbindins in renal tissues, the functional importance of this hormone in calbindin synthesis and other normal mole-rat physiology is not known.

Animals↗

PubMed1993

Tissue interactions with underlying dura mater inhibit osseous obliteration of developing cranial sutures.

Cranial sutures play a critical role in calvarial morphogenesis, serving as growth centers during skull development. Both biomechanical tensile forces originating in the cranial base and biochemical factors present in dura mater have been postulated as determinants of suture morphogenesis and patency. A rat transplant model free of the putative biomechanical influence of the dura and cranial base was used to investigate the role of the dura mater in both the initial morphogenesis and maintenance of sutures during skull growth. Day 19 fetal presumptive (F19) and day 1 neonatal differentiated (N1) coronal sutures, including associated frontal and parietal bones, were transplanted with or without underlying dura mater to the center of adult parietal bones. After 1, 2, and 3 weeks, transplanted tissues were examined histologically and histomorphometrically to determine whether sutures formed and whether they were obliterated by ossification in the absence of dura mater. Both F19 and N1 sutures remained patent for 2 weeks either in the presence or the absence of transplant dura mater. However, at 3 weeks, in the absence of transplant dura mater, sutures were obliterated by bone, while in the presence of dura mater sutures resisted ossification, demonstrating an essential requirement for interactions with the transplant dura mater in maintaining functional sutures. Both F19 and N1 transplants showed comparable bone growth (cross-sectional surface area), regardless of the presence of transplant dura mater. These experiments suggest that tissue interactions of a biochemical nature, rather than biomechanical forces generated through the cranial base, are required to maintain the suture as a non-ossified growth center. Furthermore, while the presence of dura mater was essential for maintenance of suture patency, fetal dura mater was not required for initial suture formation.

Animals↗

PubMed1992

Noncytoplasmic and filamentous appearance of calbindin-D28k and tubulin in double, indirect immunofluorescent staining of embryonic chick tissue.

Double indirect immunofluorescent labeling of embryonic chick tissue was undertaken for the vitamin D-induced calcium binding protein, calbindin-D28k, and microtubules. Immunoreactivities for both calbindin-D28k and tubulin were found to exhibit a filamentous staining pattern in mesonephros, metanephros, intestine, and brain. In the intestine, staining was absent at 19 days, while immunolabeling of tubulin became evident at 20 days, and both antigens were present in 21-day tissue. In intestinal epithelium, as well as in 10-day metanephros, it was strikingly evident that cells either stained for both antigens or were negative for both calbindin-D28k and tubulin. In 11-12-day metanephros, an increased number of cells with both immunoreactivities were found. In 15-17-day brain, tubulin was evident within all cells but stained most intensely in Purkinje cells which were also positive for calbindin-D28k. Mesonephros from 4-5-day embryos revealed immunolabeling of both tubulin and the calcium binding protein. A statistical analysis of the various cell types revealed that the vast majority contained either both antigens or neither of the immunoreactivities. Of the more than 600 cells scored, none were found to be positive for calbindin-D28k, while at the same time negative for tubulin. It is concluded that calbindin-D28k exhibits a noncytoplasmic distribution in all tissues tested and that the filamentous appearance may reflect localization of the antigen in tubulo-vesicular organelles associated with cytoskeleton.

Animals↗

PubMed1992

Estrogen inhibits calbindin-D28k expression in mouse uterus.

The cellular localization and hormonal controls of calbindin-D9k expression in the rodent reproductive tract have suggested new functions for this protein. The present studies were undertaken to extend the earlier studies of calbindin-D9k to the related protein, calbindin-D28k. Immunohistochemical studies revealed that calbindin-D28k was absent from female rat reproductive tissues, but was abundantly expressed in immature mouse uterus and oviduct. Immunoreactivity was restricted to the endometrial and glandular epithelium of the uterus and the oviductal epithelium. Neither 1,25-dihydroxyvitamin D- nor strontium-containing diets (to blunt 1,25-dihydroxyvitamin D production) affected expression of calbindin-D28k. Uterine, but not oviductal, calbindin-D28k decreased markedly at sexual maturity; this pattern persisted in pregnant mice and was reproduced in immature mice by the administration of estradiol (3 micrograms/day for 3 days). RNA extraction and Northern analyses demonstrated that estrogen markedly decreased calbindin-D28k mRNA abundance in the uterus, but not in the oviduct. These findings suggest that estrogen affects mammalian calbindin-D28k expression and represent a rare example of estrogen-induced down-regulation of gene expression.

Animals↗