Biomedical subjects

D N Kalu

Publications and source records attributed to D N Kalu.

At least 37 records · Page 2Linked to original sources

PubMed1999

The effects of gonadectomy on bone size, mass, and volumetric density in growing rats are gender-, site-, and growth hormone-specific.

Peak volumetric bone mineral density (BMD) is determined by the growth in bone size relative to the mineral accrued within its periosteal envelope. Thus, reduced peak volumetric BMD may be the result of reduced mineral accrual relative to growth in bone size. Because sex steroids and growth hormone (GH) influence bone size and mass we asked: What are the effects of gonadectomy (Gx) on bone size, bone mineral content (BMC), areal and volumetric BMD in growing male and female rats? Does GH deficiency (GH-) reduce the amount of bone in the (smaller) bone, i.e., reduce volumetric BMD? Does GH- alter the effect of Gx on bone size and mineral accrual? Gx or sham surgery was performed at 6 weeks in GH- and GH replete (GH+) Fisher 344 male and female rats. Changes in bone size, volume, BMC, areal and volumetric BMD, measured using dual X-ray absorptiometry (DPX-L), were expressed as percentage of controls at 8 months (mean +/- SEM). All results shown were significant (p < 0.05 level) unless otherwise stated. In GH+ and GH- males, respectively, Gx was associated with: lower femur volume (24%, 25%), BMC (43%, 45%), areal BMD (21%, 14%), and volumetric BMD (30%, 28%); lower spine (L1-L3) volume (26%, 28%), BMC (26%, 30%), and areal BMD (28%, 12%), but not volumetric BMD. Following Gx, GH+ females had increased femur volume (11%), no effect on BMC, decreased areal BMD (6%) and decreased volumetric BMD (17%); GH- females had no change in femur volume, but decreased femur BMC (24%), areal BMD (10%), and volumetric BMD (25%). In GH+ and GH- females, respectively, Gx was associated with a decrease in spine (L1-L3) BMC (12%, 15%), areal BMD (16%, 15%), and volumetric BMD (10%, 16%) with no change in volume. Deficits in non-Gx GH- relative to non-Gx GH+ (males, females, respectively) were: femur BMC (49%, 37%), areal BMD (23%, 8%), volume (19%, 19%) and volumetric BMD (37%, 22%); spine (L1-L3) BMC (46%, 42%), areal BMD (37%, 43%), volume (10%, 15%), and volumetric BMD (40%, 33%). Testosterone and GH are growth promoting in growing male rats, producing independent effects on bone size and mass; deficiency produced smaller appendicular bones with reduced volumetric BMD because deficits in mass were greater than deficits in size. At the spine, the reduction in size and accrual were proportional, resulting in a smaller bone with normal volumetric BMD. In growing female rats, estrogen was growth limiting at appendicular sites; deficiency resulted in a GH-dependent increase in appendicular size, relatively reduced accrual, and so, reduced volumetric BMD in a bigger bone. At the spine, accrual was reduced while growth in size was normal, thus volumetric BMD was reduced in the normal sized bone. Understanding the pathogenesis of low volumetric BMD requires the study of the differing relative growth in size and mass of the axial and appendicular skeleton in the male and female and the regulators of the growth of these traits.

Animals↗

PubMed1998

Growth hormone increases vertebral and femoral bone strength in osteopenic, ovariectomized, aged rats in a dose-dependent and site-specific manner.

The aim of the study was to assess the effect of growth hormone (GH) as restorative therapy in an aged, ovariectomized rat model with established osteopenia. The study was planned as a dose-response study, and four different skeletal sites were investigated by mechanical testing and measurements of bone mass and dimensions. Twelve-month-old virgin F344 rats were divided into eight groups with ten animals per group: (1) sham operated (sham); (2) ovariectomized (ovx); (3) sham + solvent vehicle (sv); (4) ovx + sv; (5) ovx + GH 50 microg/kg body weight/day; (6) ovx + GH 1.25 mg/kg body weight/day; (7) ovx + GH 2.5 mg/kg body weight/day; and (8) ovx + GH 5.0 mg/kg body weight/day. Groups 1 and 2 were killed after 3 months to establish that bone loss had occurred due to ovx. One month later, the remaining groups began 3 months of treatment, at the end of which the animals were also killed. The effects of ovariectomy (ovx) and GH therapy were measured at four skeletal sites: lumbar vertebrae; femoral diaphysis; femoral neck; and distal femoral metaphysis. Ovariectomy induced a significant loss of bone strength at all sites apart from the femoral neck. The loss was most pronounced at the distal femoral metaphysis. GH was able to reverse the ovx-induced loss of strength at the vertebral site in a dose-dependent manner. At the femoral diaphyseal site, GH not only reversed the ovx-induced changes but increased load values significantly above sham level. However, at the distal femoral metaphysis, which is dominated by cancellous bone, only partial reversal was seen after GH treatment. The lowest GH dose had no significant effect at any site tested. We conclude that GH treatment can reestablish vertebral bone loss due to ovariectomy in a dose-dependent manner. The restorative effect is only partial at the distal femoral metaphysis even at a high dose. At skeletal sites with less pronounced ovx-induced bone loss (femoral neck and diaphysis), GH treatment increased bone strength to sham level or above sham level. Therefore, the effect of ovariectomy is dependent upon the skeletal site investigated, and the effect of GH treatment is dependent on both the skeletal site and the size of the ovx-induced bone loss at this site.

Aging↗

PubMed1998

Aged-rodent models of long-term growth hormone therapy: lack of deleterious effect on longevity.

Studies were carried out to examine the effects of long-term recombinant human growth hormone (GH) therapy on longevity in rodents. In the first study, 150 18-month-old female F344 rats were divided into three groups of 50 rats per group: Group 1, solvent vehicle; Group 2, 10 microg GH/kg body weight three times per week; Group 3, 50 microg GH/kg body weight three times per week. GH and solvent vehicle therapies were started at 18 months of age and continued until all the animals died spontaneously. Serum insulin-like growth factor (IGF)-I was measured at 18 and 29 months of age and on 3-month-old rats. Serum IGF-I level decreased between 3 and 29 months of age. GH therapy reversed the decrease in a dose-dependent manner, with the 50 microg GH dose returning the serum IGF-I level to that of 3-month-old animals. However, statistical analysis revealed no significant effect of GH therapy on median life span, 10th percentile life span, or maximum life span. Similar observations on longevity were made on aged F344 male rats and on aged Balb/c mice, even when the dose of GH was increased to 1.0 mg/kg body weight two times per week. The main pathologic lesions in control animals were nephropathy, cardiomyopathy, leukemia, and testicular interstitial cell tumor; the prevalence of these lesions was not significantly altered by GH therapy. We conclude that long-term low-dose GH therapy that includes doses in the range that is given to humans in clinical trials in GH deficiency and to revert age-related physiologic declines has no overt deleterious effects on longevity and pathology in aged rodents.

Aging↗

PubMed1998

Modulation of intestinal estrogen receptor by ovariectomy, estrogen and growth hormone.

Ovarian hormone deficiency decreases and estrogen (E2) and growth hormone (GH) administrations increase intestinal absorption of calcium (Ca++). However, the underlying mechanisms are uncertain. To examine whether alterations in the binding characteristics of intestinal estrogen receptors (ERs) are involved, we developed and validated methods for simultaneous measurement of intestinal ERs in cytosolic and nuclear fractions and applied these techniques to four groups of female rats: sham-operated, ovariectomized (Ovx), Ovx + 5 micrograms E2/kg b.wt./day and Ovx + 8 mg GH/kg. b.wt./day. All animals were killed on day 21, and mucosal cells harvested from the duodenum for ER determination. The cytosolic and nuclear ERs were 117.2 +/- 2.7 fmol/mg protein and 64.9 +/- 1.2 fmol/mg DNA, respectively, in sham-operated rats and decreased by 16.1% and 17.0% to 98.4 +/- 1.7 fmol/mg protein and 53.8 +/- 1.3 fmol/mg DNA, respectively in Ovx rats (P < .001). E2 therapy prevented completely the decrease in cytosolic and nuclear ERs that occurred in Ovx rat (126.1 +/- 2.9 fmol/mg protein and 68.0 +/- 3.0 fmol/mg DNA, respectively, in the E2-treated group). Similarly, GH administration prevented the decrease in cytosolic and nuclear ERs that resulted from ovariectomy (119.2 +/- 3.2 fmol/mg protein and 63.4 +/- 1.3 fmol/mg DNA, respectively, in the GH-treated group). The Kd of nuclear ER-ligand complex was 2.0 +/- 0.03 nM in sham-operated rats and was slightly modulated by Ovx, E2 and GH (3.3 +/- 0.02, 2.33 +/- 0.09 and 2.23 +/- 0.04 nM, respectively, P < .001), but the Kd of cytosolic ER-ligand complex was not altered by Ovx, E2 or GH. Our findings indicate that E2 deficiency down-regulates, whereas E2 and GH administrations up-regulate intestinal ERs and prevent ovariectomy-induced decrease in receptor binding affinity. We conclude that E2 deficiency, E2 and GH may modulate intestinal Ca++ absorption, in part, by altering the abundance and binding characteristics of intestinal ERs.

Animals↗

PubMed1997

Modulation of intestinal vitamin D receptor by ovariectomy, estrogen and growth hormone.

Age-related decline in intestinal calcium (Ca) absorption often occur in postmenopausal osteoporotic women. The impaired Ca absorption can be corrected by estrogen (E2) therapy. Growth hormone (GH) therapy has also been reported to increase intestinal absorption of calcium. Since 1,25-dyhydroxyvitamin D (1,25(OH)2D) is the primary regulator of calcium absorption, we explored whether the mechanisms by which E2 and GH enhance Ca absorption involves the vitamin D endocrine system. We measured serum 1,25(OH)2D concentrations and determined the binding characteristics of intestinal vitamin D receptors (VDRs) in four groups of female rats: sham operated (sham), ovariectomized (ovx), ovx + E2, and ovx + GH. Serum 1,25(OH)2D levels were 42.4 +/- 3.4 and 42.5 +/- 3.2 pg/ml in sham and ovx rats, respectively, and decreased by 63 and 34% (P < 0.001) in ovx + E2 and ovx + GH-treated rats, respectively. The numbers of total, unoccupied and occupied VDRs were 116.9 +/- 2.0, 72.1 +/- 1.1 and 44.8 +/- 1.9 fmol/mg protein, respectively, in sham operated rats, and decreased significantly following ovariectomy by 24, 27 and 19% (P < 0.01), respectively. E2 therapy not only significantly increased total, unoccupied and occupied VDRs above those of ovx rats by 55, 58 and 49% respectively, but it increased the levels above those of sham operated controls as well (P < 0.01). In contrast, GH administration prevented the decrease that occurred in ovx rats in the number of total and unoccupied VDRs (111.2 +/- 3.3; 72.6 +/- 1.4 fmol/mg protein, respectively), but it had no significant effect on the number of occupied VDRs. The dissociation constant (Kd) of intestinal VDRs was unaltered by ovariectomy, E2 and GH. We conclude that down regulation of intestinal VDRs may contribute to the Ca malabsorption that occurs in ovarian hormone deficient states such as postmenopausal osteoporosis, and that the stimulation of Ca absorption by E2 and GH may result, in part, from up regulation of intestinal VDRs.

Animals↗

PubMed1997

Effects of age and food restriction on calcium signaling in parotid acinar cells of Fischer 344 rats.

In this study, we characterized alpha-adrenergic (alpha AR) and muscarinic induced [Ca2+]i changes in individual parotid acinar cells from male Fischer 344 rats (6-24 month-old) fed ad libitum (AL) or 60% ad libitum intake (FR). Cells were prepared by collagenase/hyaluronidase digestion. [Ca2+]i was measured by video image, fluorescent microscopy in single acinar cells loaded with FURA2. Neither age nor food restriction altered the peak [Ca2+]i achieved in response to carbachol (100 microM). Similar results were obtained for epinephrine (Epi = 100 microM) stimulation in 6- and 12-month-old animals. However, the peak [Ca2+]i response to Epi declined between 12 and 18 months in both dietary groups (e.g., AL: 12 months = 387 +/- 21 nM, 18 months = 253 +/- 10 nM; FR: 12 months = 430 +/- 22 nM, 18 months = 325 +/- 14 nM). The decline in response to Epi seen with age was less in FR than in AL animals at 18 months, but not at 24 months. In addition, db cAMP reduced the carbachol-stimulated [Ca2+]i response to levels comparable to those observed with epinephrine. The results support the view that calcium mobilization in parotid acinar cells from male Fischer 344 rats in response to alpha AR, but not to muscarinic, stimulation is impaired with age. Food restriction may slow down, but does not prevent, the functional decline. Furthermore, cAMP appears to modulate the muscarinic response.

Aging↗

PubMed1997

Effects of aging and food restriction on alpha 1-adrenergic receptors and G-protein in the rat parotid gland.

The aim of this study is to investigate age-related changes in alpha 1-adrenergic receptor and G-protein in rat parotid gland, and to examine whether the age-related changes are modulated by food restriction. The binding characteristics of alpha 1-adrenergic receptor were measured in parotid cell membranes from 3-, 6-, 12-, 18-, and 24-month-old ad libitum-fed (AL) and food-restricted (FR) rats. The number of alpha 1-adrenergic receptors (Bmax) increased between 3 and 6 months by 24.5% and 16.2% in AL and FR rats, respectively, with no further significant change up to 24 months of age. The Kd values of [3H]prazosin binding increased by 53.6% in AL rats, but decreased by 16.1% in FR rats between 3 and 6 months, and the changes were maintained up to 24 months of age. The binding activity of G-protein with [35S]guanosine 5'-[gamma-Thio]triphosphate (GTP[gamma S]) in parotid cell membranes was measured in 6- and 24-month-old AL and FR rats. Basal GTP binding capacity decreased by 22.3% and 26.0% in AL and FR rats, respectively, between 6 and 24 months. Epinephrine-stimulated increase in GTP binding capacity decreased by 19.2% between 6 and 24 months in AL rats, but not in FR rats. The Kd values of [35S]GTP[gamma S] binding were not affected by aging or food restriction. In summary: (1) FR decreased maturation-related increase in the number of alpha 1-adrenergic receptors and prevented maturation-related decrease in receptor binding affinity between 3 and 6 months of age. However, between 6 and 24 months of age FR had no further significant effect on the number of alpha 1-adrenergic receptors and on receptor binding affinity. (2) FR prevented age-related decrease in alpha 1-adrenergic agonist-stimulated increase in GTP binding capacity. We conclude that FR may have a modulatory effect on age-related impairment in alpha 1-adrenergic receptor function resulting from altered G-protein binding activity in the rat parotid gland.

Aging↗

PubMed1997

Modulation by food restriction of intracellular calcium signaling in parotid acinar cells of aging Fischer 344 rats.

Previous studies suggest that alpha 1-adrenergic (alpha 1-AR)-induced intracellular calcium ([Ca2+]i) mobilization in rat parotid acinar cells declines with age. In this study, we examined the effects of food restriction on alpha 1-AR and muscarinic-stimulated [Ca2+]i mobilization in parotid acinar cells during aging. [Ca2+]i levels in response to the alpha 1-AR agonist epinephrine and the muscarinic agonist carbachol were evaluated in Fura-2-loaded parotid acinar cells from ad libitum-fed (AL) and food-restricted (FR) Fischer 344 male rats at 4, 6, 14, and 24 months of age. [Ca2+]i responses to epinephrine and carbachol (10 microM) were significantly reduced (48% and 35%, respectively; p < .05) in cells from 24-month-old AL rats as compared to younger AL rats. In contrast, no significant reduction of epinephrine and carbachol responses was observed in 24-month-old FR animals. An age-related increase in basal [Ca2+]i (peak around 14 months; p < .02) was observed in both AL and FR rats. In addition, basal [Ca2+]i was higher in FR than in AL rats at 14 and 24 months of age (p < .02). These studies suggest that FR partially attenuates or delays age-related impairments in alpha 1-AR- and muscarinic-cholinergic signal transduction systems of parotid acinar cells. Basal [Ca2+]i also appears to be altered during aging and by FR.

Adrenergic Agonists↗

PubMed1996

Putative intestinal estrogen receptor: evidence for regional differences.

Previously we demonstrated that 17 beta-estradiol (E2) stimulates intestinal calcium absorption and enhances uptake of calcium by intestinal mucosal cells in vitro, and that the latter contains estrogen receptor (ER)-like proteins and mRNAs. The current study was undertaken to further characterize the putative intestinal ERs using RT-PCR analysis, Western blot analysis, Southern blot analysis, ligand binding assays and gel shift assays. RT-PCR analysis using primers directed at the terminal end of the A/B domain of the uterine ER confirmed our previous finding that intestinal ER mRNAs are present throughout the intestine, with the exception that the duodenum lacked the amplified region. Western blot analysis, using ER-715 antibody, detected the expected 68- to 70-kDa ER protein and additional 58-, 46- and 41-kDa proteins in the uterus and colon, while duodenal extract contained only a faint 46-kDa and strong 28-kDa protein bands. Southern blot analysis performed on enzyme-digested genomic DNAs demonstrated the presence of ER-like sequences in genomic DNA from the uterus, duodenum and colon. However, enzyme restriction maps of genomic DNAs from intestinal segments were different from the map for uterine genomic DNA digested with similar enzymes. In ligand binding assays, 125I-labeled E2 bound specifically to 46-kDa protein in duodenal, colonic and uterine extracts and the binding was competitively inhibited by excess cold E2. Gel mobility shift assays using 32P-labeled vitellogenin derived ERE demonstrated that jejunal, colonic and uterine ER protein contain the classical DNA binding domain. In addition, ERE bound in a specific fashion to sites in genomic DNAs from the uterus, colon and jejunum. In contrast, protein extracts and genomic DNAs from the duodenum failed to bind to 32P-labeled ERE and, consequently, appear to be devoid of the classical DNA binding domain. These findings establish the presence of ER-like proteins and genes in intestinal mucosal cells of rats and suggest that the duodenum contains a variant ER gene that encodes a variant ER protein.

Animals↗

PubMed1995

Evolution of the pathogenesis of postmenopausal bone loss.

At a time when the investigation of bone diseases appears to be on the threshold of a new era, it is appropriate to pause and contemplate past and present thinking on the etiology of postmenopausal osteoporosis. This brief review traces the evolution of ideas on the nature of the pathogenesis of this disease from Albright's hypoosteoblastic hypothesis through other hypotheses that include disturbance in osteoclasia, negative calcium balance, disturbance of calcium homeostatic control mechanisms, increased skeletal sensitivity to parathyroid hormone, deficiency of calcitriol and calcitonin, altered activities of growth factors and cytokines, alterations in the local regulation of osteoclastogenesis, and changes in mechanical usage set points.

Bone Resorption↗

PubMed1994

In vivo effect of 17 beta-estradiol on intestinal calcium absorption in rats.

Previously we reported that intestinal cells contain estrogen receptors, and that 17 beta-estradiol enhanced calcium uptake by these cells in vitro. The current study was undertaken to examine the in vivo effects of 17 beta-estradiol on intestinal absorption of calcium and phosphorus. Three groups of rats were studied. Group 1 received solvent vehicle. Groups 2 and 3 received 5 micrograms and 40 micrograms 17 beta-estradiol/kg body weight/day, respectively, for 21 days. Hormone and solvent vehicle injections were given subcutaneously. Rats were fed a Teklad diet containing 0.4% Ca, 0.3% P and 3.0 U vitamin D/g during the study. Intestinal absorption of calcium and phosphorus was assessed over a 5-day period from day 15-19. Carmine red (25 mg/100 g diet) was added to the rat feed to mark the beginning and end of fecal collections. Administration of 17 beta-estradiol caused an increase in intestinal absorption of calcium and phosphorus. The increase was significant only for calcium, and in the animals that received high-dose 17 beta-estradiol (P < 0.05). Serum calcium and phosphorus levels were significantly greater in 17 beta-estradiol treated than in control animals. The urinary excretion of calcium and phosphorus was also increased in a dose-dependent manner by 17 beta-estradiol, and was significant for both calcium and phosphorus in animals that received high-dose 17 beta-estradiol (P < 0.05). In contrast, 17 beta-estradiol treatment did not significantly alter the serum levels of parathyroid hormone and 1,25(OH)2 vitamin D. These findings indicate that estrogen administration promotes intestinal absorption of calcium in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Effects of ovariectomy and estrogen on the serum levels of insulin-like growth factor-I and insulin-like growth factor binding protein-3.

To determine the effects of ovariectomy and 17 beta-estradiol (E2) on serum IGF-I and its binding proteins, female Sprague-Dawley rats, aged 95 days, were divided into four groups. Group 1 was sham-operated; groups 2, 3, and 4 were ovariectomized. Groups 3 and 4 received daily injections of 200 ng (low dose) and 5000 ng (high dose) E2/kg body wt./day, respectively and the others were given solvent vehicle. Ovariectomy resulted in a significant increase in serum IGF-I (P < 0.001) at 30 and 35 days post-surgery; the increase was prevented in animals that received low-dose E2 while high-dose E2 reduced serum IGF-I levels below those of the sham-operated controls (P < 0.01). Serum IGF-binding proteins (IGFBPs) were determined by IGF-ligand blot analysis, and the resulting autoradiograms quantified by laser densitometry. The intensity of the IGFBP-3 bands changed in parallel with serum IGF-I levels. Ovariectomy increased, low-dose E2 restored, and high-dose E2 reduced serum IGFBP-3 levels compared to the levels for the sham-operated controls. The intensities of binding protein bands smaller than those of IGFBP-3 appeared unchanged by the treatment regimens. A Western immunoblot analysis with IGFBP-3 antiserum confirmed the ligand-blot data. The changes in the levels of IGF-I and its binding proteins were accompanied by ovariectomy-induced increase in osteoblast and osteoclast numbers and loss of cancellous bone that were attenuated by E2 administration. We conclude that there is a possible role for IGF-I in the pathogenesis of the increased bone turnover that occurs early in ovarian hormone deficiency.

Animals↗

PubMed1993

Evaluation of the molecular and cellular basis for the modulation of thyroid C-cell hormones by aging and food restriction.

Male F344 rats fed ad libitum or maintained on 60% of the ad libitum food intake were sacrificed at 6, 12, 18 and 24 months of age. The thyroids were removed for the analysis of the C-cell hormones, calcitonin, calcitonin gene-related peptide (CGRP) and somatostatin. In the animals fed ad libitum, the peptide content of all three hormones and their mRNA pools increased significantly with age. The increases were markedly suppressed by food restriction. Similarly, the rate of mRNA synthesis of the hormones increased with age and was attenuated by food restriction. Calcitonin and CGRP containing cells increased in number with age in the ad libitum fed animals. In the food restricted animals the numbers of calcitonin positive cells were consistently but not significantly lower than those of ad libitum fed animals at similar ages. In the case of CGRP containing cells, their numbers were significantly lower in the food restricted than in the ad libitum fed animals from 18 months of age. Our findings indicate that aging and food restriction modulate the levels of the thyroidal C-cell hormones at the levels of cell proliferation and possibly gene transcription.

Aging↗

PubMed1993

Ovariectomy-induced bone loss and the hematopoietic system.

To investigate the relationship of the hematopoietic system to the loss of bone due to ovarian hormone deficiency, we examined the effects of ovariectomy and estrogen administration on the thymus, spleen and the bone marrow, and on the proliferation of marrow progenitors of osteoclasts. We also assessed the effects of daily administration of interleukin-1 receptor antagonist (IL-1ra) on bone loss due to ovarian hormone deficiency. Ovariectomy resulted in decreased cancellous bone volume, increased trabecular osteoblast and osteoclast numbers, and increased serum alkaline phosphatase levels that were prevented by 17 beta-estradiol treatment. Thymus weight, spleen weight, thymus and spleen lymphocytes, and bone marrow monocytes and lymphocytes also increased significantly following ovariectomy, and the increases were suppressed by 17 beta-estradiol. Ovariectomy, in addition, caused a 4-fold increase in the number of tartrate resistant acid phosphatase (TRAP)-positive multinucleated cells formed in cultures of marrow cells and the increase was partially inhibited by 17 beta-estradiol. IL-1ra administration did not prevent the bone loss due to ovariectomy. Our findings indicate that ovariectomy-induced bone loss in the rat is accompanied by marked changes in the hematopoietic system, and that these changes are modulated by estrogen administration. In spite of the negative finding with IL-1ra, the nature of the involvement of the hematopoietic system in the pathogenesis of bone loss due to ovarian hormone deficiency merits continued exploration.

Acid Phosphatase↗

PubMed1993

In vivo effects of transforming growth factor-beta 2 in ovariectomized rats.

In vitro studies indicate that transforming growth factor-beta (TGF-beta) has a role in the regulation of bone cell activities. However, little is known about the effects of TGF-beta on bone when it is administered systemically. This study was undertaken to evaluate the in vivo effects of TGF-beta 2 on bone and marrow cells in the ovariectomized rat bone loss model. Female Sprague-Dawley rats, aged 95 days, were divided into 4 groups. Group 1 was sham operated; groups 2-4 were ovariectomized. Groups 3 and 4 received daily injections of 10 micrograms and 50 micrograms of TGF-beta 2/kg body weight, respectively. Groups 1 and 2 received the solvent vehicle. All animals were sacrificed after 35 days. Ovariectomy caused a significant increase in, total mononuclear marrow cells, the number of TRAP positive multinucleated cells formed in culture of marrow cells, and the number of trabecular osteoclasts and osteoblasts. These increases were associated with loss of cancellous bone in the proximal tibia. TGF-beta 2 completely prevented the increase in the number of TRAP positive multinucleated cells, and caused a small but not statistically significant decrease in the number of trabecular osteoclasts. However, TGF-beta 2 had no significant effect on the number of total mononuclear marrow cells and on the loss of cancellous bone due to ovariectomy. We conclude that TGF-beta 2 probably plays a role in the regulation of the proliferation of osteoclast progenitors in bone marrow in vivo. Studies carried out over a longer period are required to determine whether it will modulate the increase in osteoclast and osteoblast numbers that occur in cancellous bone following ovariectomy.

Animals↗

PubMed1993

Estrogen modulates the mRNA levels for cancellous bone protein of ovariectomized rats.

This study was undertaken to examine the effects of ovariectomy and 17 beta-estradiol (E2) on the gene expression of type 1 collagen, osteocalcin and the protooncogen, c-myc, in cancellous bone. Female Sprague-Dawley rats, aged 95 days, were divided into 4 groups. Group 1 was sham operated and Groups 2-4 were ovariectomized. Groups 3 and 4 received daily injections of 160 ng and 1600 ng E2/kg body weight, respectively. Groups 1 and 2 received the solvent vehicle. All animals were sacrificed after 14 days. The femurs were dissected out and cancellous bone scraped from the distal metaphysis. RNA was isolated from the cancellous bone, immobilized on filters or size-fractionated by agarose gel electrophoresis and adsorbed on filters which were then hybridized with specific cDNA probes. Ovariectomy resulted in a significant increase in the mRNAs of type 1 collagen, osteocalcin and c-myc. The increase was suppressed in animals that received 17 beta-estradiol injections. In addition, ovariectomy caused the expected decrease in cancellous bone in the proximal tibia and increased osteoclast and osteoblast numbers. The ovariectomy-induced changes were prevented by 17 beta-estradiol administration. These findings suggest that the lack of ovarian hormones shortly after ovariectomy up-regulates and estrogen administration down-regulates the expression of important cancellous bone matrix proteins as well as the protooncogen, c-myc.

Animals↗

PubMed1993

Evidence for estrogen receptor-linked calcium transport in the intestine.

Intestinal calcium malabsorption in postmenopausal osteoporotic women is often linked indirectly to decreased serum 1,25(OH)2 vitamin D or to intestinal resistance to its action, rather than directly to the low circulating estrogen that results following menopause. The studies presented indicate that the intestinal mucosal cells of rats contain estrogen receptor immunoreactivity, express the mRNA for estrogen receptors, and respond directly to 17 beta-estradiol with enhanced calcium transport that is suppressed by gene transcription and protein synthesis inhibitors. These findings suggest that estrogen has a physiological role in the regulation of intestinal calcium absorption and that its deficiency in postmenopausal osteoporosis, and following therapeutic oophorectomy, may result directly in calcium malabsorption that is believed to be an important factor in the bone loss that occurs in these conditions.

Animals↗

PubMed1992

Effects of aging and food restriction on the trigeminal ganglion: a morphometric study.

A quantitative morphometric study of the rat trigeminal ganglion was conducted to determine the changes that occur with aging. All measurements were tracked from young to old age in two rat groups simultaneously. One group was fed ad libitum, the other was maintained on restricted food intake from 6 weeks on. Immunocytochemical and radioimmunoassay techniques were used to study the neuron group that produces the peptide, CGRP and to compare it with the CGRP-negative neuron group. We observed that in the trigeminal ganglion, soma diameters and nucleus diameters of all neurons, whether CGRP positive or negative, increased modestly with age; so did total ganglion weight. Food restriction delayed, but did not prevent the increases in neuron diameters. No significant changes occurred as a function of age in the total number of neurons per ganglion, the ratio of CGRP positive to CGRP negative neurons and ganglion content of CGRP. Food restriction did not affect the parameters that remained constant with age. These findings are in contrast to the marked inhibitory effect of food restriction on age-related increase in thyroid calcitonin, a hormone that is encoded by the same gene as CGRP.

Aging↗