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PubMed1982

A new method for achieving complete two-layer closure of a massive palatal cleft.

A new surgical technique was used to provide a complete two-layer closure of an extremely wide cleft palate in cases in which the width of the cleft defect is larger than the sum of the combined widths of the remaining palate. The oral layer of closure is obtained by the use of turnover flaps of the nasal mucoperiosteum based on the margin of the cleft. This recruits the nasal mucosal flap into the oral layer of closure and allows adequate width for complete closure of the oral layer of the cleft. The nasal layer of closure is obtained by the use of a wide, long, superiorly based pharyngeal flap. This technique provides, in a single surgical procedure, a complete two-layer closure of extremely wide palatal defects. Previously, staged procedures were necessary to obtain a complete two-layer closure, or if a single procedure was performed, an incomplete two-layer closure resulted.

Adolescent↗
PubMed1996

Egg jelly layers of Xenopus laevis are unique in ultrastructure and sugar distribution.

Jelly coats surrounding the eggs of the South African clawed toad, Xenopus laevis, consist of three transparent, gelatinous layers: the innermost layer (J1), the middle layer (J2), and the outer layer (J3). The distribution of N-acetylglucosamine within these jelly coats, as probed with FITC-conjugated wheat germ agglutinin (WGA-FITC), and the matrix ultrastructure of each layer, as visualized in platinum replicas produced by the quickfreeze, deep-etch, and rotary-shadowing technique, suggests that each layer has a unique fiber and glycoprotein composition. J1 extends nearly 200 microns from the egg surface and exhibits no WGA-FITC staining. Stereo images of platinum replicas indicates that J1 consists of a tightly knit network of 5-10 nm fibers decorated with 10-20 nm particulate components. In contrast, J2 is a relatively thin layer, extending only 25-40 microns from the outer aspect of J1. When visualized by confocal microscopy, J2 displays a multilayered WGA-FITC staining pattern. The ultrastructure of J2 consists of sheets of fine fibers that run parallel to one another and that can be identified by their ability to bind WGA-colloidal gold. The fibers of each sheet run at an oblique angle to fibers in neighboring layers. J3 extends 100 microns or more from J2. The WGA-FITC staining pattern shows high intensity in its outer region and less intensity in regions closer to J2. Like J1, the J3 ultrastructure consists of a network of 5-10 nm fibers, decorated with 10-20 nm particulate components. The results of these studies add to a growing body of information that suggests the jelly coats surrounding the eggs of many animals consist of a fibrous glycoprotein superstructure that acts as a scaffold to which globular glycoproteins are bound.

Animals↗
PubMed2000

Single versus two layer suturing for closing the uterine incision at caesarean section.

BACKGROUND: A single layer closure might save operating time, disrupt less tissue, introduce less foreign material into the wound, and thus result in less post operative morbidity. OBJECTIVES: The objective of this review was to assess the effects of a single layer closure of the uterine incision at caesarean section, in contrast to conventional two layer closure. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials of single layer closure, either interrupted or continuous, compared to two layer closure with continuous sutures in pregnant women undergoing elective or emergency caesarean section. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Two trials involving 1006 women were included. Based on one trial, single layer closure was associated with reduced operating time (5.6 minutes). Based on one trial, radiographic scar appearance showed fewer scar defects at three months with the single closure group. There were no statistically significant differences in the use of extra haemostatic sutures, incidence of endometritis, decrease in post operative haematocrit or use of blood transfusion. REVIEWER'S CONCLUSIONS: There appear to be no advantages or disadvantages for routine use of single layer closure compared to two layer closure, except perhaps a shorter operation time.

Cesarean Section↗
PubMed1989

SEM characterization of a cellular layer separating blood vessels from endoderm in the quail embryo.

The associations between the developing blood vessels and both endoderm and splanchnic mesoderm in quail embryos at stages 9-11 were examined by using scanning electron microscopy. Embryos were pinned ventral-side up on agar plates and the endoderm was surgically removed prior to fixation and dehydration. This procedure exposes a netlike layer of cells closely apposed to the ventral surface of paraxial mesoderm and all visible blood vessels; we are calling this the subvascular layer. Development of this layer proceeds rostral-to-caudal, and lateral-to-medial, with the earliest stages of formation being visible over the unsegmented paraxial mesoderm of the segmental plate. The subvascular layer increases markedly in density slightly medial to the innermost boundary of the intraembryonic vascular plexus. Cells of this layer eventually establish a continuous sheet beneath the lateral plate and paraxial mesoderm and the notochord. With maturation, the cells of the subvascular layer approach confluence. The spatial and temporal patterns of development of the embryonic vascular tissues and the subvascular layer are closely correlated, suggesting a possible role for the subvascular layer in normal embryonic vascular development.

Animals↗
PubMed1993

Determination of the induced ELF electric field distribution in a two layer in vitro system simulating biological cells in nutrient solution.

In-vitro studies of biological effects of electromagnetic fields are often conducted with cultured cells either in suspension or grown in a monolayer. In the former case, the exposed medium can be assumed to be homogeneous; however, eventually the cells settle to the bottom of the container forming a two layer system with different dielectric and conductive properties. In the present work the effect of this separation on the electric field distribution is calculated and experimentally measured at selected positions for a commonly used exposure configuration. The settled cell suspension is modeled by a well-defined two layer system placed in a rectangular container with the base of the container parallel to the direction of the magnetic field. Theoretical calculations based on numerical techniques are done for various two layer systems with different conductivities in each layer. The agreement between the theoretical calculations and the experimental measurements is within +/- 1.5 mV/m, or 10% of the maximum induced field when the conductivity of the lower layer is ten times that of the upper layer. This result is well within experimental error. When the thickness of one of the layers is small compared to the thickness of the other layer, it is found that the electric field distribution is essentially that of the homogeneous case. The latter situation corresponds to a typical cell exposure condition.

Cells, Cultured↗
PubMed2002

Controlled enzymatic removal of damaging casein layers on medieval wall paintings.

A new, gentle enzymatic method was developed for a controlled removal of casein layers from medieval wall paintings. These casein layers were applied over the last 60 years on wall paintings in order to decrease substantial damage due to a peeling off of the frescoes from the roughcast surface due to environmental effects. However, due to the aging of the casein layers (at 40-50 years), a more drastic peeling occurred and the danger of total destruction of the wall paintings is severe. Thus, screening was performed to find the most suitable enzyme for casein digestion. Alcalase 2.5 DX L was the most appropriate enzyme for an effective proteolysis reaction. The enzyme was immobilized on functionalized cellulose membrane. A membrane pad system with immobilized enzymes was developed which could be pressed on the casein layers on the wall painting. A controlled removal of the casein layers by proteolytic digestion was observed and it was possible to continuously wash off the hydrolyzed casein fragments from the wall painting surface by an aqueous carbonate buffer flowing through the membrane pad. The removal and the digestion was monitored by reverse HPLC. Additionally, an on-line monitoring system was set up in order to continuously follow the casein layer removal and the digestion procedure directly on the wall painting. This technique is based on noninvasive 2D-fluorescence monitoring. Optical fiber systems were used to continuously monitor the fluorescence intensity of casein-bound tryptophan. The off-line data were verified with the on-line 2D-fluorescence data. Based on the scientific result an appropriate technique for the controlled enzymatic removal of damaging casein layers on the surface of medieval wall paintings using immobilized enzyme is now available. It is now applied to remove such casein layers from medieval wall paintings in the Allerheiligen-Kapelle Cloister, Wienhausen, Germany, and the St. Alexander Kirche, Wildeshausen, Germany.

Caseins↗
PubMed2006

A bridge between pillared-layer and helical structures: a series of three-dimensional pillared coordination polymers with multiform helical chains.

Rational self-assembly of a long V-shaped 3,3',4,4'-benzophenonetetracarboxylate (bptc) ligand and metal salts in the presence of linear bidentate ligand yield a series of novel pillared helical-layer complexes, namely, [Cu2(bptc)(bpy)2] (1), [M3(Hbptc)2(bpy)3(H2O)4].2 H2O (M = Fe(2) and Ni(3)), [Co2(bptc)(bpy)(H2O)].0.5 bpy (4), [Cd2(bptc)(bpy)(H2O)2].H2O (5), [Mn2(bptc)(bpy)1.5(H2O)3] (6) and [M2(bptc)(bpy)0.5(H2O)5].0.5 bpy (M = Mn(7), Mg(8) and Co(9), bpy=4,4'-bipyridine). Their structures were determined by single-crystal X-ray diffraction analyses and further characterized by elemental analyses, IR spectra, and thermogravimetric (TG) analyses. The structure of 1 consists of two types of chiral layers, one left-handed and the other right-handed, which are connected by bpy pillars to generate a novel 3D open framework featuring four distinct helical chains. Compounds 2 and 3 are isostructural and feature 3D structures formed from the interconnection of arm-shaped helical layers with bpy pillars. Compound 4 is a pillared helical double-layer complex containing four different types of helices, among which the nine-fold interwoven helices constructed from triple-stranded helical motifs are unprecedented. Compound 5 exhibits a novel 3D covalent framework which features nanosized tubular channels. These channels are built from helical layers pillared by bptc ligands. The structure of 6 is constructed from {Mn(bptc)(H2O)}n2n- layers, which consist of left- and right-handed helical chains, pillared by [Mn2(bpy)3(H2O)4]4+ complexes into a 3D framework. To the best of our knowledge, compounds 1-6 are the first examples of pillared helical-layer coordination polymers. Compounds 7-9 are isostructural and exhibit interesting 2D helical double-layer structures, which are constructed from {M(bptc)(H2O)2}n2n- ribbons cross-linked by [M2(bpy)(H2O)6]4+ complexes. Furthermore, the 3D supramolecular structures of 7-9 are similar to the 3D structure of 6, and the 2D structure of 7 can be transformed into the 3D structure of 6 at higher reaction temperature. By inspection of the structures of 1-9, it is believed that the V-shaped bptc ligand and V-shaped phthalic group of the bptc ligand are important for the formation of the helical structures. The magnetic behavior of compounds 1, 2, 4, 6, and 9 was studied and indicated the existence of antiferromagnetic interactions. Moreover, compound 5 shows intense photoluminescence at room temperature.

Journal Article↗
PubMed2001

Thalamic terminal morphology and distribution of single corticothalamic axons originating from layers 5 and 6 of the cat motor cortex.

We investigated the axonal morphology of single corticothalamic (CT) neurons of the motor cortex (Mx) in the cat thalamus, using a neuronal tracer, biotinylated dextran amine (BDA). After localized injection of BDA into the Mx, labeled CT axons were found ipsilaterally in the thalamic reticular nucleus (TRN), the ventroanterior-ventrolateral complex (VA-VL), the central lateral nucleus (CL), the central medial nucleus, and the centromedian nucleus, but with the primary focus in the VA-VL. The terminals in the VA-VL formed a large laminar cluster, which extended approximately in parallel with the internal medullary lamina. The laminar organization mirrored morphologic features of single CT axons. We reconstructed the trajectories of 25 single CT axons that arose from layer V (16 axons) or layer VI (9 axons) and terminated in the VA-VL. Terminals of single CT axons that originated from both layer V and layer VI were confined within a laminar structure about 700 microm thick, suggesting the existence of laminar input organization in the VA-VL. Otherwise, the two groups of the CT axons showed contrasting features. All of the CT axons derived from layer VI gave rise to a few short collaterals to the TRN and then formed extensive arborization with numerous small, drumstick-like terminals in the VA-VL. On the other hand, the CT axons arising from layer V gave rise to collaterals whose main axons descended into the cerebral peduncle. Each collateral projected to the VA-VL or CL without projection to the TRN and formed a few small clusters of giant terminals. The two groups of CT neurons in the same cortical column had convergent rather than segregated termination in the VA-VL. However, the terminals of layer VI CT neurons were distributed diffusely and widely in the VA-VL, whereas the terminals of layer V CT neurons were much more focused and surrounded by the terminals of the former group. These contrasting features of the two types of CT projections appear to represent their different functional roles in the generation of motor commands and control of movements in the Mx.

Animals↗
PubMed1985

Development of the human retina: patterns of cell distribution and redistribution in the ganglion cell layer.

Neurogenesis in the ventricular layer and the development of cell topography in the ganglion cell layer have been studied in whole-mounts of human fetal retinae. At the end of the embryonic period mitotic figures were seen over the entire outer surface of the retina. By about 14 weeks gestation mitosis had ceased in central retina and differentiation of photoreceptor nuclei was evident within a well-defined area which constituted about 2% of total retina area. This area was approximately centered on the site of the putative fovea, identified by the exclusive development of cone nuclei at that location. The area of retina in which mitosis had ceased increased as gestation progressed. By mid-gestation mitosis in the ventricular layer occupied about 77% of the outer surface of the retina and by about 30 weeks gestation mitosis in the ventricular layer had ceased. Cell density distributions in the ganglion cell layer were nonuniform at all stages studied (14-40 weeks). Densities were highest at about 17 weeks gestation, and by mid-gestation the adult pattern of cell topography was present with maps showing elevated cell densities in posterior retina and along the horizontal meridian. Cell densities generally declined throughout the remainder of the gestation period, except in the posterior retina, where densities in the perifoveal ganglion cell layer remained high during the second half of gestation. There is a rapid decline in cell density in the foveal ganglion cell layer toward the end of gestation, and it is suggested that the persistence of high densities in the perifoveal region may be related to migration of cells away from the developing fovea. The total population of cells in the ganglion cell layer was highest (2.2-2.5 million cells) between about weeks 18 and 30 of gestation. After this the cell population declined rapidly to 1.5-1.7 million cells. It is suggested that naturally occurring neuronal death is largely responsible for this decline.

Cell Count↗
PubMed1991

Distributions of choline acetyltransferase and acetylcholinesterase activities in the retinal layers of the red-tailed hawk and road runner.

The activities of choline acetyltransferase and acetylcholinesterase were assayed in submicrogram samples from layers of red-tailed hawk and road runner retina. Both enzyme activities were concentrated in and near the inner plexiform layer. Within the inner plexiform layers of both species, activities of each enzyme were concentrated in two bands, one in each half of this layer. Little choline acetyltransferase activity was found superficial to the middle third of the inner nuclear layer. The distributions of acetylcholinesterase activities corresponded well to those of choline acetyltransferase, except in the outer plexiform layer and the outer margin of the inner nuclear layer of the hawk. These distributions of enzyme activities indicate that populations of amacrine cells in the retinae of these species are cholinergic. In addition to these same cells and presumably cholinoceptive amacrine and ganglion cells, acetylcholinesterase activity in the hawk was associated with a population of horizontal cells that may be unrelated to synaptic cholinergic neurotransmission. Choline acetyltransferase activities associated with amacrine somata and processes were about four times greater in the hawk than in the road runner, suggesting important differences in the density and function of cholinergic elements between species. Possible synaptic relationships in the inner plexiform layer consistent with the interspecies differences in enzyme activities are considered.

Acetylcholinesterase↗
PubMed2006

Preparation and characterisation of dual-layer latex-coated columns for open-tubular capillary electrochromatographic preconcentration of cations combined in-line with their separation by capillary electrophoresis.

A dual-layer ion-exchange latex-coated column was prepared and characterised for on-capillary preconcentration of cations using an open-tubular ion-exchange CEC format. After preconcentration, the analyte cations were eluted with a transient isotachophoretic gradient and separated by CE. The latex double layer was established by first coating the negatively charged wall of the capillary with a layer of cationic quaternary ammonium anion-exchange Dionex AS5A latex particles (60 nm diameter), and then coating a layer of anionic sulphonated cation-exchange Dionex CS3 latex particles (300 nm diameter) onto the underlying AS5A layer. The adhesion of layers is based on electrostatic attractions. Several dual-layer capillaries were characterised for their EOF and ion-exchange capacity and this showed that coatings could be prepared reproducibly by a simple flushing procedure. The dual-layer columns exhibited a moderate, pH-independent EOF (ca. 26 x 10(-9 )m2V(-1)s(-1)) and an ion-exchange capacity of 57 microequiv./g (or 2.69 nequiv./column). Using an 8 cm length of coated capillary combined with a 72 cm length of untreated capillary, a method for on-line preconcentration and separation of monovalent organic bases, alkali metal ions and alkaline earth metal ions by CE was developed. Recoveries for the preconcentration step were 48% for 4-methylbenzylammonium, 43% for benzylammonium, 30-32% for alkali metal ions and 71-75% for alkaline earth cations. In all cases, recoveries were reproducible with RSDs being less than 6.2%. The influences of the ion-exchange selectivity coefficient of the analyte and the sample-loading rate on analyte recovery were also examined. The proposed method was utilised for the determination of alkaline earth cations and low microM detection limits were obtained.

Benzylammonium Compounds↗
PubMed2002

Improved localization of coronary stents using layer decomposition.

Accurate placement and expansion of coronary stents is hindered by the fact that most stents are only slightly radiopaque and hence difficult to see in typical coronary X-ray images. We propose a new technique for improved image guidance of multiple coronary stent deployment using layer decomposition of coronary X-ray image sequences. We hypothesize that layer decomposition can improve the accuracy of localization of the end of a deployed stent. Layer decomposition is used to obtain good quality images of a stent in vitro. The resultant background-subtracted stent images are embedded into other cine X-ray image sequences to form a database of simulated image sequences. For each simulated sequence, the position of the stent edge is estimated from raw and layer-decomposed images using a small region of the original layer image as a template. Layer decomposition reduced median position errors in 33 of 47 image sequences (70%), including 16 of 18 sequences in which the position errors for raw and layer images differed by 5.0 pixels (0.5 mm) or more. Layer decomposition significantly reduces errors in determination of stent edge location in simulated cine X-ray image sequences.

Cineangiography↗
PubMed2004

Microstructure of MmM(5)/Mg multi-layer hydrogen storage films prepared by magnetron sputtering.

Multi-layer hydrogen storage thin films with Mg and MmNi3.5(CoAlMn)1.5 (here Mm denotes La-rich mischmetal) as alternative layers were prepared by direct current magnetron sputtering. Transmission electron microscopy investigation shows that the microstructure of the MmNi3.5(CoAlMn)1.5 and Mg layers are significantly different although their deposition conditions are the same. The MmNi3.5(CoAlMn)1.5 layer is composed of two regions: one is an amorphous region approximately 4 nm thick at the bottom of the layer and the other is a nanocrystalline region on top of the amorphous region. The Mg layer is also composed of two regions: one is a randomly orientated nanocrystalline region 50 nm thick at the bottom of the layer and the other is a columnar crystallite region on top of the nanocrystalline region. These Mg columnar crystallites have their [001] directions parallel to the growth direction and the average lateral size of these columnar crystallites is about 100 nm. A growth mechanism of the multi-layer thin films is discussed based on the experiment results.

Journal Article↗
PubMed2004

Chemical and thermal denaturation of crystalline bacterial S-layer proteins: an atomic force microscopy study.

Crystalline monomolecular cell surface layers, S-layers, are one of the most common outermost cell envelope components of the prokaryotic organisms (bacteria and archaeda) that protects them from competitive habitats. Since isolated S-protein subunits are able to re-assemble into crystalline arrays on lipid films and solid supports making biomimetic surfaces, S-layer technology is currently used in nanobiotechnology. An important aspect of the biomimetic surfaces built with S-layers is their stability under extreme solvent conditions or temperature. Chemical (pH, alcohol) and physical (thermal) denaturant conditions were employed to test the stability of S-layers. Recrystallized bacterial surface layers from Bacillus sphaericus (SbpA) on hydrophilic silicon wafers loses the crystalline structure at 80% ethanol/water mixtures, the change in structure being reversible after treating the surface with buffer solution. SbpA on silicon supports denatures at pH 3 and at 70 degrees C, and the process is irreversible. Cross-linking of SbpA enhances the stability for high ethanol and acidic conditions, but it does not improve thermal stability. Recrystallized SbpA on secondary cell wall polymer (SCWP), a natural environment for the protein layer, is more resistant to ethanol and pH exposure than recrystallized SbpA on hydrophilic silicon supports. Atomic force microscopy (AFM) was used to monitor the loss of stability and the changes in protein layer conformation.

Bacillus↗
PubMed2006

Intravascular hematocrit layering in equilibrium phase contrast-enhanced MR angiography of the peripheral vasculature.

PURPOSE: To evaluate intravenous hematocrit (HCT) layering detected in patent calf veins that potentially simulate filling defects seen in deep venous thrombosis (DVT). MATERIALS AND METHODS: Flow characteristics, vessel size, and enhancement patterns of veins with HCT layering were examined and compared with those of normal veins in vivo. Ex vivo studies were conducted with conventional and blood pool contrast agents to study the effects of MRI parameters and intrinsic variables on detecting HCT layering. RESULTS: HCT layering was detected in high resolution equilibrium phase images in one of six healthy subjects, and five of five patients with peripheral vascular occlusive disease (PVOD). Layering occurred only in deep veins that had significantly slower flow and larger diameter (P < 0.001) compared to normal veins. Enhancement patterns observed in vivo were similar to those found in the ex vivo study. CONCLUSION: The HCT layering effect in MR venography has not been previously reported, likely because delayed equilibrium phase venous imaging is not frequently performed. The use of blood pool contrast agents and high resolution equilibrium phase imaging allows intravascular HCT layering to be observed within a 30-60-minute time frame. Several imaging characteristics that distinguish HCT layering from DVT are discussed.

Adult↗
PubMed2004

Different physiological MRI noise between cortical layers.

Significantly higher temporal fluctuations of the blood oxygenation level-dependent (BOLD) signal in the living rat group compared to that in the dead rat group were observed in the cortex, suggesting the existence of physiological information in the signal fluctuations. A similar analysis shows significantly different fluctuations between visual cortical layers. The highest fluctuations were observed in layers 4 and 5 and the lowest in layer 1. Given the consistency with published electrophysiology studies anticipating high spontaneous activity in the deeper layers (particularly layer 4), and low activity in superficial layers, we hypothesize that the BOLD signal temporal fluctuations may reflect cortical neuronal activity. Temporal fluctuations in ultrahigh spatial resolution data of the rat brain were measured in two ways. In the first, analyses were performed according to known layer widths, and in the second equal lines of 117 micro along the cortex were selected. The second approach yielded temporal fluctuations along the cortex that resemble known neuronal density distributions including the intralayer structure, particularly within layer 5.

Animals↗
PubMed1997

Cell adhesion molecules regulate guidance of dorsal root ganglion axons in the marginal zone and their invasion into the mantle layer of embryonic spinal cord.

In order to elucidate the mechanisms regulating the projections of dorsal root ganglion (DRG) axons in the dorsal funiculus and invasion into target regions in the mantle layer (prospective gray matter) of the spinal cord, we examined the interactions between DRG axons and spinal cord. DRG neurons were dissociated from chick embryos and cultured for 1-2 days on cryostat sections of the spinal cord at embryonic day 5 (E5) or at E9. E5 and E9 DRG neurons extended neurites onto both marginal zone (prospective white matter) and mantle layer (prospective gray matter) of the spinal cord, suggesting that both of these regions are permissive for neurite growth. When E5 DRG neurites approached cryosections of E5 spinal cord from outside, most of them ran in the marginal zone without invading the mantle layer. In contrast, about half of E9 DRG neurites entered the mantle layer after crossing the marginal zone of E9 spinal cord. These growth patterns of DRG neurites on spinal marginal zone and mantle layer are similar to the pathway formation of DRG axons at comparable stages in vivo; DRG axons run exclusively in the prospective dorsal funiculus before E6, and enter the mantle layer (prospective dorsal horn) to reach the target regions by E9. Perturbation of functions of Ng-CAM, Nr-CAM, and axonin-1/SC2 by adding the specific antibodies in the culture medium increased the ratio of DRG neurites entering the mantle layer of E5 spinal cord, suggesting that these cell adhesion molecules are involved in keeping DRG neurites in the marginal zone. Taken together with the expression of Ng-CAM, Nr-CAM, and axonin-1/SC2, these CAMs on DRG axons may regulate the guidance of these axons in the marginal zone before E6, and the subsequent decrease in the relative levels of these CAMs might allow DRG axons to invade the target mantle layer.

Animals↗
PubMed1997

Expression of messenger ribonucleic acids of luteinizing hormone and follicle-stimulating hormone receptors in granulosa and theca layers of chicken preovulatory follicles.

Expression of luteinizing hormone receptor (LHR) and follicle-stimulating hormone receptor (FSHR) mRNAs was demonstrated in the granulosa and theca layers of the large preovulatory follicles of the chicken ovary by Northern hybridization and reverse transcription-polymerase chain reaction (RT-PCR). Northern hybridization results showed multiple LHR and FSHR mRNA transcripts and the predominant species were 2.3 and 2.5 kb, respectively. The highest abundance of LHR mRNA was found in the granulosa layer of the largest follicle (F1 follicle), while the abundance remained low in the granulosa layers of the third (F3) and fifth largest (F5) follicles. FSH mRNA abundance was the highest in the granulosa layer of F5 follicle, but decreased in the granulosa layers of F3 and F1 follicles. In the theca layers of all the three follicles examined LHR and FSHR mRNAs were extremely low. These results were confirmed by RT-PCR experiment which involved coamplification of LHR or FSHR mRNA and beta-actin mRNA as the internal control. The LHR PCR product was sequenced and indicated 92.2% homology with the corresponding region of the quail LHR cDNA. This study indicated that the marked increase in expression of LHR mRNA in granulosa layer of the F1 follicle might be important for LHR protein synthesis and succeeding bonus progesterone production in F1 follicle destined to ovulation. However, higher expression of FSHR mRNA in the granulosa layer of the less mature follicles may be involved in the differentiation and maturation of granulosa cells in these follicles.

Animals↗