FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W1974865240> ?p ?o ?g. }

• W1974865240 endingPage "1670" @default.
• W1974865240 startingPage "1661" @default.
• W1974865240 abstract "To develop a model for the study of physiological angiogenesis, we transplanted ovarian follicles onto striated muscle tissue and analyzed the process of microvascularizationin vivo using repeated fluorescence microscopy. Follicles were mechanically isolated from unstimulated as well as pregnant mare’s serum gonadotropin (PMSG)- or PMSG/luteinizing hormone (LH)-stimulated Syrian golden hamster ovaries and were transplanted as free grafts into dorsal skinfold chambers of untreated or synchronized hamsters. Follicles lacking thecal cell layers did not vascularize regardless whether harvested from unstimulated or PMSG-stimulated animals, but underwent granulosa cell apoptosis, as indicated in vivo by nuclear condensation and fragmentation of bisbenzimide-stained follicular tissue. In contrast, all follicles at 48 hours after PMSG treatment with a multilayered thecal shell exhibited initial signs of angiogenesis within 3 days. Vascularization was completed within 7 to 10 days, comprising a dense glomerulum-like microvascular network. Nature and extent of vascularization of follicles harvested at 72 hours after either PMSG or PMSG/LH treatment did not notably differ from each other when transplanted into the respective synchronized animals. However, follicles with PMSG/LH treatment revealed significantly larger microvessel diameters and higher capillary blood perfusion compared to follicles with sole PMSG treatment, probably reflecting the adaptation to the increased functional demand upon the LH surge. Using the unique experimental approach of ovarian follicle transplantation in the dorsal skinfold chamber of Syrian golden hamsters, we could show in vivo the developmental stage-dependent vascularization of follicular grafts with sustained potential to meet their metabolic demand by increased blood perfusion. To develop a model for the study of physiological angiogenesis, we transplanted ovarian follicles onto striated muscle tissue and analyzed the process of microvascularizationin vivo using repeated fluorescence microscopy. Follicles were mechanically isolated from unstimulated as well as pregnant mare’s serum gonadotropin (PMSG)- or PMSG/luteinizing hormone (LH)-stimulated Syrian golden hamster ovaries and were transplanted as free grafts into dorsal skinfold chambers of untreated or synchronized hamsters. Follicles lacking thecal cell layers did not vascularize regardless whether harvested from unstimulated or PMSG-stimulated animals, but underwent granulosa cell apoptosis, as indicated in vivo by nuclear condensation and fragmentation of bisbenzimide-stained follicular tissue. In contrast, all follicles at 48 hours after PMSG treatment with a multilayered thecal shell exhibited initial signs of angiogenesis within 3 days. Vascularization was completed within 7 to 10 days, comprising a dense glomerulum-like microvascular network. Nature and extent of vascularization of follicles harvested at 72 hours after either PMSG or PMSG/LH treatment did not notably differ from each other when transplanted into the respective synchronized animals. However, follicles with PMSG/LH treatment revealed significantly larger microvessel diameters and higher capillary blood perfusion compared to follicles with sole PMSG treatment, probably reflecting the adaptation to the increased functional demand upon the LH surge. Using the unique experimental approach of ovarian follicle transplantation in the dorsal skinfold chamber of Syrian golden hamsters, we could show in vivo the developmental stage-dependent vascularization of follicular grafts with sustained potential to meet their metabolic demand by increased blood perfusion. In adult tissue, angiogenesis is a characteristic of pathological conditions, such as tumor growth,1Folkman J Tumor angiogenesis.Adv Cancer Res. 1985; 43: 174-203Google Scholar, 2Folkman J Angiogenesis in cancer, vascular, rheumatoid and other disease.Nat Med. 1995; 1: 27-31Crossref PubMed Scopus (7153) Google Scholar wound healing,3Greenburg GB Hunt TK Proliferative response in vitro of vascular endothelial and smooth muscle cells exposed to wound fluids and macrophages.J Cell Physiol. 1978; 97: 353-360Crossref PubMed Scopus (122) Google Scholar, 4Polverini PJ Cotran PS Gimbrone MA Unanue ER Activated macrophages induce vascular proliferation.Nature. 1977; 269: 804-806Crossref PubMed Scopus (610) Google Scholar, 5Banda MJ Knighton DR Hunt TK Werb Z Isolation of a nonmitogenic angiogenic factor from wound fluid.Proc Natl Acad Sci USA. 1982; 79: 7773-7777Crossref PubMed Scopus (150) Google Scholar, 6Dvorak HF Tumors: wounds that do not heal. Similarities between tumor stroma generation and wound healing.N Engl J Med. 1986; 315: 1650-1659Crossref PubMed Scopus (3442) Google Scholar and inflammation.7Jones MK Wang H Peskar BM Levin E Itani RM Sarfeh IJ Tarnawski AS Inhibition of angiogenesis by nonsteroidal anti-inflammatory drugs. Insight into mechanisms and implications for cancer growth and ulcer healing.Nat Med. 1999; 5: 1418-1423Crossref PubMed Scopus (790) Google Scholar Nonpathological angiogenesis is rare and restricted to the female reproductive tissues.8Reynolds LP Killilea SD Redmer DA Angiogenesis in the female reproductive system.FASEB J. 1992; 6: 886-892Crossref PubMed Scopus (363) Google Scholar During the cycle of follicle development and corpus luteum formation, vascular changes are tightly regulated in that angiogenesis is turned on for brief periods and then completely inhibited. Thus, folliculogenesis offers a unique system to study not only the induction of angiogenesis, but also the maturation and regression of blood vessels.9Goede V Schmidt T Kimmina S Kozian D Augustin HG Analysis of blood vessel maturation processes during cyclic ovarian angiogenesis.Lab Invest. 1998; 78: 1385-1394PubMed Google Scholar The precise control of angiogenesis in the developing ovarian follicle and corpus luteum is critical for normal reproductive function. The regulators of this physiological angiogenesis, however, are not yet completely elucidated. Preovulatory follicles in mammalian ovaries develop from the vast pool of preantral follicles. During their development preantral follicles grow through successive stages10Tonetta SA diZerega GS Intragonadal regulation of follicular maturation.Endocr Rev. 1989; 10: 205-229Crossref PubMed Scopus (156) Google Scholar in which they differ in size, number of granulosa cell layers, and absence or presence of thecal cells.11Roy SK Greenwald GS An enzymatic method for dissociation of intact follicles from the hamster ovary: histological and quantitative aspects.Biol Reprod. 1985; 32: 203-215Crossref PubMed Scopus (91) Google Scholar Some 50 years ago, Bassett12Bassett DL The changes in the vascular pattern of the ovary of the albino rat during the estrous cycle.Am J Anat. 1943; 73: 251-291Crossref Scopus (137) Google Scholar described changes in the vasculature of the developing follicles and corpora lutea in rat ovaries during the estrous cycle. More recent studies have demonstrated the angiogenic potential of corpora lutea extracts,13Gospodarowicz D Thakral KK Production of a corpus luteum angiogenic factor responsible for proliferation of capillaries and neovascularization of the corpus luteum.Proc Natl Acad Sci USA. 1978; 75: 847-851Crossref PubMed Scopus (135) Google Scholar follicular fluid,14Frederick JL Shimanuki T diZerega GS Initiation of angiogenesis by human follicular fluid.Science. 1984; 224: 389-390Crossref PubMed Scopus (79) Google Scholar and granulosa cell-conditioned medium.15Rone JD Goodman AL Preliminary characterization of angiogenic activity in media conditioned by cells from luteinized rat ovaries.Endocrinology. 1990; 127: 2821-2828Crossref PubMed Scopus (17) Google Scholar These angiogenic activities have later been attributed to the action of basic fibroblast growth factor,16Neufeld G Ferrara N Schweigerer L Mitchell R Gospodarwicz D Bovine granulosa cells produce basic fibroblast growth factor.Endocrinology. 1987; 121: 597-603Crossref PubMed Scopus (145) Google Scholar some heparin-binding growth factors,17Grazul-Bilska AT Redmer DA Reynolds LP Production of heparin-binding angiogenic factor(s) by bovine corpora lutea during pregnancy.J Anim Sci. 1992; 70: 254-262Crossref PubMed Scopus (20) Google Scholar and vascular endothelial growth factor.18Kamat BR Brown LF Manseau EJ Senger DR Dvorak HF Expression of vascular permeability factor/vascular endothelial growth factor by human granulosa and theca lutein cells: role in corpus luteum development.Am J Pathol. 1995; 146: 157-165PubMed Google Scholar, 19Shweiki D Itin A Neufeld G Gitay-Goren H Keshet E Patterns of expression of vascular endothelial growth factor (VEGF) and VEGF receptors in mice suggest a role in hormonally regulated angiogenesis.J Clin Invest. 1993; 91: 2235-2243Crossref PubMed Scopus (500) Google Scholar Although it is widely recognized that angiogenesis is a prerequisite for ovarian and uterine function, only a few investigators have taken advantage of the physiological angiogenic processes in the female reproductive system to study mechanisms that underlie the induction and regulation of angiogenesis. This paucity of information is related in part to the lack of simple reproducible animal models of follicular angiogenesis. To extend our understanding of the mechanisms that regulate nonpathological vascular growth, we used the hamster dorsal skinfold chamber as the host site for ovarian follicle transplantation and systematically analyzed in vivo the host’s angiogenic response to follicular grafts using multifluorescence microscopy. The chamber preparation contains one layer of striated muscle and skin and allows for intravital microscopic observation of the microcirculation in the awake animals throughout a prolonged period of time. The chamber technique and its implantation procedure have been described previously in detail.20Endrich B Asaishi K Goetz A Messmer K Technical report—a new chamber technique for microvascular studies in unanesthetized hamsters.Res Exp Med. 1980; 177: 125-134Crossref PubMed Scopus (366) Google Scholar In brief, under pentobarbital sodium anesthesia (50 mg/kg body weight i.p.), two symmetrical titanium frames were implanted on the extended dorsal skinfold of 8- to 10-week-old Syrian golden hamsters (body weight, 60 to 80g), so that they sandwiched the double layer of skin. One layer of skin was then removed in a circular area of ∼15 mm in diameter, and the remaining layers (consisting of striated skin muscle and subcutaneous tissue) were covered with a removable coverslip incorporated into one of the titanium frames. In addition, a permanent catheter was passed from the dorsal to the ventral side of the neck and inserted into the jugular vein. After intravenous application of 0.2 ml of 5% fluorescein isothiocyanate (FITC)-labeled dextran 150,000 (Sigma, Deisenhofen, Germany), the chamber enabled for continuous observation and repetitive analysis of the microcirculation by means of intravital fluorescence microscopy in the awake animal. The animals were allowed to recover from anesthesia and surgery for at least 48 hours. For follicle donation, 8- to 10-week-old female hamsters were intraperitoneally anesthetized with pentobarbital sodium (50 mg/kg body weight). After laparotomy, donor ovaries were aseptically removed and placed in 30-mm-diameter Falcon plastic Petri dishes filled with 37°C warm Dulbecco’s modified Eagle’s medium (10% fetal calf serum, 0.1 mg/ml gentamicin), and the fluorescent vital dye bisbenzimide H33342 (200 μg/ml; Sigma). After removing the surrounding tissue, the ovaries were microdissected under a stereo microscope using 27-gauge needles. According to size, the follicles were visually collected and transferred into 37°C warm bisbenzimide H33342-free Dulbecco’s modified Eagle’s medium (Figure 1A). For follicle transplantation, the cover glass of the dorsal skinfold chamber was removed and one to three follicles of either size were placed on the striated muscle within the chamber (Figure 1B). A handpicking procedure guaranteed single connective tissue-free follicles for transplantation. In general, follicles were grouped in accordance to their initial size at the time point of harvesting and transplantation with 1) diameters <250 μm, 2) diameters in the range of 250 to 500 μm, and 3) diameters >500 μm. Two donor animals were pretreated with pregnant mare’s serum gonadotropin (PMSG, Sigma) dissolved in phosphate-buffered saline (1000 U/ml). PMSG was given subcutaneously at 8 a.m. in the morning at a single dose of 2 U/10 g body weight. Follicles were harvested at either 48 hours or 72 hours after the single PMSG treatment. In parallel, skinfold chamber-equipped age-matched female hamsters (n = 13) were also treated by a single subcutaneous injection of PMSG (2 U/10 g body weight) and follicles were transplanted at either 48 hours or 72 hours after PMSG treatment. One donor animal was pretreated with PMSG as described above, followed by subcutaneous application of luteinizing hormone (LH) (25 μg/hamster; Sigma) at 8 a.m. in the morning of day 2 after application of PMSG. Follicles were harvested 24 hours after the LH application. In parallel, skinfold chamber-equipped age-matched female hamsters (n = 7) were also treated by PMSG followed by LH. Follicles harvested from an animal with neither PMSG nor LH treatment were used for transplantation into nontreated skinfold chamber-equipped age-matched female hamsters (n = 6). For in vivo microscopic observation, the awake animals were immobilized in a Plexiglas tube and the skinfold preparation was attached to the microscopic stage. The stage was placed on a computer-controlled microscope desk, which allowed repeated scanning of each individual follicle for intravital microscopy. Intravenous injection of 0.2 ml of 5% FITC-labeled dextran 150,000 (Sigma) guaranteed contrast enhancement by staining of the plasma. Rhodamine 6G (0.1%, 0.1 ml i.v.; Sigma) allowed for the direct in vivo staining of leukocytes. Intravital microscopy was performed using a modified Leitz Orthoplan microscope with a 100 W HBO mercury lamp attached to a Ploemo-Pak illuminator with blue, green, and ultraviolet filter blocks (Leitz, Wetzlar, Germany) for epi-illumination. The microscopic images were recorded by a charge-coupled device video camera (CF8/1 FMC; Kappa GmbH, Gleichen, Germany) and transferred to a video system for off-line evaluation. With the use of ×4, ×6.3, ×10, and ×20 long distance objectives (Leitz), magnifications of ×86, ×136, ×216, and ×432 were achieved on a 14-inch video screen (PVM 1444; Sony, Tokyo, Japan). Quantitative off-line analysis of the videotapes was performed by means of a computer-assisted image analysis system (CapImage, Zeintl, Heidelberg) and included the determination of the diameter (μm) and the size of the transplanted follicles (mm2), the size of the growing microvascular networks (in percentage of the follicular size), the microvessel density, ie, the length of red blood cell (RBC)-perfused microvessels per observation area (cm/cm2), and the diameters of the follicular microvessels (μm). On ultraviolet epi-illumination the dye bisbenzimide is characterized by a bright blue fluorescence with only little bleaching that persists through several cell generations. The specific fluorescence/background fluorescence ratio is high enough throughout a period of 3 weeks to precisely delineate the stained follicular graft from the surrounding unaffected host tissue. The area of fully developed microvascular networks might sometimes slightly exceed the follicular tissue area with the consequence that values of the size of the growing microvascular networks are >100% of the follicular size. Centerline RBC velocity (VRBC) in the individual microvessels was measured by frame-to-frame analysis. Volumetric blood flow (VQ) of individual microvessels was calculated from VRBC and diameter (D) for each microvessel as VQ = π × (D/2)2Folkman J Angiogenesis in cancer, vascular, rheumatoid and other disease.Nat Med. 1995; 1: 27-31Crossref PubMed Scopus (7153) Google Scholar × VRBC/K, where K (=1.3) represents the Baker/Wayland factor,21Baker M Wayland H On-line volume flow rate and velocity profile measurement for blood in microvessels.Microvasc Res. 1974; 7: 131-143Crossref PubMed Scopus (379) Google Scholar considering the parabolic velocity profile of blood in microvessels. Rhodamine 6G-stained leukocytes were classified in accordance to their interaction with the endothelium of newly formed microvessels. Rolling cells were defined as cells moving with a velocity less than two-fifths of the centerline velocity (given as percentage of nonadherent leukocytes passing through the observed vessel segment within 20 seconds). Adherent cells were defined as cells that did not move or detach from the endothelial lining during an observation period of 20 seconds (given as number of cells per microvascular network area). A total of 26 follicles were harvested from the nontreated donor animal and were transplanted into the skinfold chambers of six nontreated female hamsters. A total of 14 follicles at 48 hours and 10 follicles at 72 hours after PMSG treatment were transplanted into the skinfold chambers of five and eight female synchronized hamsters. A total of nine follicles at 72 hours after PMSG/LH treatments were transplanted onto the skinfold chambers of seven female synchronized hamsters. The macroscopic appearance of the skinfold chamber preparations and the implanted grafts were documented daily. Intra-vital multifluorescence microscopic analysis of growth, angiogenesis, and microcirculation was performed on days 3, 5, 7, 10, and 14 after follicular graft transplantation. Measurements of vascular density and microhemodynamic parameters included only newly formed microvessels that could be clearly distinguished by their glomerulum-like arrangement from the autochthonous host striated muscle microvessels, displaying the typical parallel arrangement of the muscle capillaries.22Menger MD Lehr HA Scope and perspectives of intravital microscopy—bridge over from in vitro to in vivo.Immunol Today. 1993; 14: 519-522Abstract Full Text PDF PubMed Scopus (149) Google Scholar Vascular density was measured in five regions of interest per graft and observation time point. Microvascular diameters as well as hemodynamic parameters were determined from 10 microvessels per region of interest. At the end of the in vivo experiments, ie, day 14 after follicle transplantation, the animals were sacrificed with an overdose of pentobarbital, and the skinfold chamber preparations were processed for light microscopic analysis. Follicles were harvested from ovaries of untreated and PMSG- and PMSG/LH-treated hamsters and grouped according to size, fixed in formalin (4% in phosphate-buffered saline) for 24 hours at 4°C, embedded in paraffin, serially sectioned, and stained with hematoxylin and eosin (H&E). Sections through the central plane of the follicle (ie, those containing the largest cross-sectional area) were videotaped, and cross-sectional areas of the individual follicles were measured by computer-assisted planimetry for their staging.11Roy SK Greenwald GS An enzymatic method for dissociation of intact follicles from the hamster ovary: histological and quantitative aspects.Biol Reprod. 1985; 32: 203-215Crossref PubMed Scopus (91) Google Scholar Statistical analysis was performed using a general linear mixed model with log-transformed values of the parameters vascularized area, microvessel density, microvessel diameter, red blood cell velocity, and volumetric blood flow as dependent variables. Confidence intervals were calculated by fitting a linear model to log-transformed data that in turn were retransformed into original scales (error bars in Figure 5 and 6). Accordingly, summary statistics are expressed as geometric means and 95% confidence intervals (lower/upper). Factors included into this nested design model were time with five levels, treatment with four levels, time by treatment interactions, host animal nested within treatment group, and follicle nested within host animal. The differences between follicles of 250 to 500 μm and >500 μm at 48 hours after PMSG as well as follicles at 72 hours after either PMSG or PMSG/LH were tested across all time points by fitting a model without the time by treatment interaction effect. P values give the results of F statistics and are not adjusted for multiplicity. Additionally, comparisons were performed at the individual time points after follicle transplantation when indicated by the comparisons across time. In that case, the interaction effect was included into the model. Calculations were performed using the restricted maximum likelihood method and the small sample correction for standard errors according to Kenward Rogers as provided by the SAS procedure mixed (SAS Institute, Cary, NC). Differences in take rate of follicles were analyzed using the Fisher exact test (SigmaStat; Jandel Corp., San Rafael, CA). The criterion for significance was taken to beP < 0.05. First, we analyzed growth, angiogenesis and microcirculation of follicles in dependency of their stage of development. Experiments were undertaken in nontreated animals and in animals after stimulation with PMSG for 48 hours. Follicles with diameters <250 μm [mean geometric diameter, 100 μm (51/161, ie, lower and upper 95% confidence intervals)], which were harvested from nontreated animals, did not vascularize. Follicles with diameters <250 μm [mean geometric diameter, 185 μm (154/242, ie, lower and upper 95% confidence intervals)], but harvested from PMSG-stimulated hamsters (48 hours) and transplanted into synchronized animals, also failed to induce neovascularization. In contrast, all follicles with diameters >250 μm established a complete microvascular network, regardless of whether they had been harvested from stimulated (n = 14) or unstimulated animals (n = 3). This was reflected by a take rate of 100% of follicles >250 μm in diameter, contrasting the take rate of 0% of follicles with a diameter <250 μm (P< 0.05). Follicles <250 μm were characterized by H&E histology as tri- to quadrilaminar follicles without any trace of theca cell layers (Figure 2A), indicating secondary follicles of stage 3 to 5 according to the classification of Roy and Greenwald.11Roy SK Greenwald GS An enzymatic method for dissociation of intact follicles from the hamster ovary: histological and quantitative aspects.Biol Reprod. 1985; 32: 203-215Crossref PubMed Scopus (91) Google Scholar Follicles with a diameter >250 μm exhibited several layers of granulosa cells with beginning antrum formation and a well-developed multilayered thecal shell (Figure 2B), reflecting stages of follicular development >6.11Roy SK Greenwald GS An enzymatic method for dissociation of intact follicles from the hamster ovary: histological and quantitative aspects.Biol Reprod. 1985; 32: 203-215Crossref PubMed Scopus (91) Google Scholar In case of follicle vascularization, initial angiogenesis was characterized by sinusoidal sacculations, capillary budding, and sprout formation. Throughout the following days protrusion of sprouts was observed, leading to interconnection of individual sprouts with a growing microvascular network (Figure 3). Finally, follicular grafts presented with a complete glomerulum-like microvascular network (Figure 3). The development of sprouts originated in >80% from the host striated muscle capillaries. Sprouts also developed from postcapillary venules of the host tissue, but only rarely from arterioles (Figure 4A). Blood flow from the follicular grafts was consistently drained by a microvascular system, which consisted of former capillary vascular segments of the host striated muscle (Figure 4B).Figure 4Intravital fluorescence microscopic images of the microvasculature of follicular grafts at day 14 after transplantation into hamster dorsal skinfold chambers. A: High magnification reveals the interaction of the newly formed microvessels with the microvasculature of the host tissue, demonstrating an arteriole (white arrows) that serves as vascular supply and multiple intercapillary anastomoses (black arrows) between the follicular capillaries (asterisk) and the striated muscle capillaries (arrowheads).B: Blood from the follicular graft is almost completely drained by a postcapillary vessel (arrowheads), which may function as a venule, but represents a former striated muscle capillary, as clearly indicated by the parallel arrangement with the other striated muscle capillaries (arrows). Blue light epi-illumination with contrast enhancement by 5% FITC-labeled dextran 150,000 i.v. Scale bars: 100 μm (A), 150 μm (B).View Large Image Figure ViewerDownload Hi-res image Download (PPT) Although no signs of angiogenesis and new vessel formation could be observed in follicles of a diameter <250 μm throughout the entire observation period of 14 days, quantitative analysis of larger follicles revealed a progressive increase of the area of vascularization with increasing microvessel density from day 3 to day 5, which then remained constant throughout the remainder of the experiment (Figure 5, A and B). Discrimination of these grafts according to size demonstrated that follicles with diameters between 250 and 500 μm showed almost comparable areas of vascularization (Figure 5A), but a tendency toward higher average values of microvessel density (Figure 5B) on days 3 and 5 compared with the >500 μm-follicles. Capillary red blood cell velocity comparably increased ∼10-fold from day 3 to day 10 in both the smaller and larger follicles (Table 1). Follicular capillary diameters were found significantly reduced from day 3 to day 10, amounting to ∼8 μm at day 14 (Table 1). Up to day 10, capillary diameters in larger follicles were markedly wider than those in smaller follicles (Table 1). Calculation of individual capillary blood perfusion in follicular grafts revealed a progressive increase of blood flow until vascularization was completed (days 7 to 10), which, however, was followed by a fall of 40 to 50% at day 14. In addition, capillaries of larger follicles showed markedly higher blood perfusion on days 5 to 14 when compared with those of smaller follicles (Table 1).Table 1Microvessel Diameter, Capillary Red Blood Cell Velocity, and Capillary Volumetric Blood Flow in Newly Developed Microvascular Networks of Follicular Grafts Upon Harvesting from Hamsters at 48 Hours after PMSG Treatment and Transplantation into Dorsal Skinfold Chambers of Synchronized AnimalsDay 3Day 5Day 7Day 10Day 14Follicles with diameters of 250 to 500 μm (mean diameter 353 μm (305/404)) Microvessel diameter (μm)14.0 (12.4/15.8)11.6 (10.3/13.1)*P < 0.05 versus day 3.10.2 (9.0/11.5)†P < 0.05versus days 3 and 5.8.5 (7.6/9.6)‡P < 0.05versus days 3, 5, and 7.8.0 (7.1/9.1)‡P < 0.05versus days 3, 5, and 7. VRBC (μm/sec)28.5 (18.9/42.8)91.6 (60.9/137.8)*P < 0.05 versus day 3.123.9 (82.4/186.4)*P < 0.05 versus day 3.220.7 (146.7/332.0)‡P < 0.05versus days 3, 5, and 7.142.2 (94.5/214.0)*P < 0.05 versus day 3. Volumetric blood flow (pL/sec)3.4 (2.2/5.2)7.4 (4.8/11.4)*P < 0.05 versus day 3.7.7 (5.0/11.9)*P < 0.05 versus day 3.9.6 (6.2/14.9)*P < 0.05 versus day 3.5.5 (3.6/8.5)Follicles with diameters >500 μm (mean diameter 750 μm (607/970)) Microvessel diameter (μm)16.0 (14.1/18.1)15.1 (13.3/17.1)¶P < 0.05versus follicles with diameters of 250 to 500 μm.11.8 (10.4/13.4)†P < 0.05versus days 3 and 5.10.6 (9.4/12.1)†P < 0.05versus days 3 and 5.¶P < 0.05versus follicles with diameters of 250 to 500 μm.8.1 (7.1/9.2)§P < 0.05versus days 3, 5, 7, and 10 VRBC (μm/sec)23.4 (15.3/35.9)103.6 (67.6/158.6)*P < 0.05 versus day 3.167.1 (109.1/255.9)*P < 0.05 versus day 3.240.0 (156.7/367.5)†P < 0.05versus days 3 and 5.213.2 (139.3/326.5)†P < 0.05versus days 3 and 5. Volumetric blood flow (pL/sec)3.6 (2.2/5.8)14.1 (8.8/22.7)*P < 0.05 versus day 3.¶P < 0.05versus follicles with diameters of 250 to 500 μm.14.0 (8.7/22.5)*P < 0.05 versus day 3.¶P < 0.05versus follicles with diameters of 250 to 500 μm.16.2 (10.1/26.1)*P < 0.05 versus day 3.8.3 (5.2/13.4)*P < 0.05 versus day 3.Values are given as geometric means with lower and upper 95% confidence intervals (number/number), which are based on a linear mixed model of log-transformed data.* P < 0.05 versus day 3.† P < 0.05versus days 3 and 5.‡ P < 0.05versus days 3, 5, and 7.§ P < 0.05versus days 3, 5, 7, and 10¶ P < 0.05versus follicles with diameters of 250 to 500 μm. Open table in a new tab Values are given as geometric means with lower and upper 95% confidence intervals (number/number), which are based on a linear mixed model of log-transformed data. In a second set of experiments, we compared the vascularization of follicles that were harvested 72 hours after either PMSG or PMSG/LH treatment and transplanted into synchronized animals. The area of vascularization ranged between 60 and 90% (days 7 to 14) without significant differences between the two cohorts of follicles (Figure 6A). Concomitantly, microvessel density increased twofold from day 3 to day 5, and then remained constant at ∼250 cm/cm2 in both the PMSG and the PMSG/LH treatment group (Figure 6B). Capillary red blood cell velocity increased significantly from <100 μm/second at day 3 to 236 to 309 μm/second at day 10 in both PMSG- and PMSG/LH-treated follicles (Table 2), similarly as observed in follicles after 48 hours of PMSG treatment. In parallel, follicular capillary diameters decreased significantly during the first 10 days after transplantation (Table 2). Capillary blood perfusion m" @default.
• W1974865240 created "2016-06-24" @default.
• W1974865240 creator A5002844010 @default.
• W1974865240 creator A5023070415 @default.
• W1974865240 creator A5045251394 @default.
• W1974865240 creator A5052617751 @default.
• W1974865240 creator A5081747456 @default.
• W1974865240 date "2001-11-01" @default.
• W1974865240 modified "2023-10-16" @default.
• W1974865240 title "In Vivo Imaging of Physiological Angiogenesis from Immature to Preovulatory Ovarian Follicles" @default.
• W1974865240 cites W103446480 @default.
• W1974865240 cites W148216960 @default.
• W1974865240 cites W1598952686 @default.
• W1974865240 cites W1963938932 @default.
• W1974865240 cites W1971728487 @default.
• W1974865240 cites W1972962741 @default.
• W1974865240 cites W1974171881 @default.
• W1974865240 cites W1976681763 @default.
• W1974865240 cites W1979826225 @default.
• W1974865240 cites W1990017270 @default.
• W1974865240 cites W1991204850 @default.
• W1974865240 cites W2000000541 @default.
• W1974865240 cites W2010833294 @default.
• W1974865240 cites W2016293999 @default.
• W1974865240 cites W2026530967 @default.
• W1974865240 cites W2026831351 @default.
• W1974865240 cites W2033377265 @default.
• W1974865240 cites W2036949899 @default.
• W1974865240 cites W2037801518 @default.
• W1974865240 cites W2052582923 @default.
• W1974865240 cites W2057106552 @default.
• W1974865240 cites W2057658664 @default.
• W1974865240 cites W2059971415 @default.
• W1974865240 cites W2062199482 @default.
• W1974865240 cites W2064508977 @default.
• W1974865240 cites W2068885726 @default.
• W1974865240 cites W2070995184 @default.
• W1974865240 cites W2077424568 @default.
• W1974865240 cites W2113787725 @default.
• W1974865240 cites W2121942269 @default.
• W1974865240 cites W2122646231 @default.
• W1974865240 cites W2156606731 @default.
• W1974865240 cites W2314203153 @default.
• W1974865240 cites W2467249203 @default.
• W1974865240 cites W2473031280 @default.
• W1974865240 cites W3149865648 @default.
• W1974865240 cites W4211002736 @default.
• W1974865240 cites W62872458 @default.
• W1974865240 doi "https://doi.org/10.1016/s0002-9440(10)63013-1" @default.
• W1974865240 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/1867040" @default.
• W1974865240 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/11696427" @default.
• W1974865240 hasPublicationYear "2001" @default.
• W1974865240 type Work @default.
• W1974865240 sameAs 1974865240 @default.
• W1974865240 citedByCount "41" @default.
• W1974865240 countsByYear W19748652402013 @default.
• W1974865240 countsByYear W19748652402014 @default.
• W1974865240 countsByYear W19748652402016 @default.
• W1974865240 countsByYear W19748652402017 @default.
• W1974865240 countsByYear W19748652402019 @default.
• W1974865240 countsByYear W19748652402021 @default.
• W1974865240 countsByYear W19748652402022 @default.
• W1974865240 crossrefType "journal-article" @default.
• W1974865240 hasAuthorship W1974865240A5002844010 @default.
• W1974865240 hasAuthorship W1974865240A5023070415 @default.
• W1974865240 hasAuthorship W1974865240A5045251394 @default.
• W1974865240 hasAuthorship W1974865240A5052617751 @default.
• W1974865240 hasAuthorship W1974865240A5081747456 @default.
• W1974865240 hasBestOaLocation W19748652401 @default.
• W1974865240 hasConcept C126322002 @default.
• W1974865240 hasConcept C134018914 @default.
• W1974865240 hasConcept C142724271 @default.
• W1974865240 hasConcept C207001950 @default.
• W1974865240 hasConcept C2777702977 @default.
• W1974865240 hasConcept C2778324911 @default.
• W1974865240 hasConcept C2780394083 @default.
• W1974865240 hasConcept C2780536345 @default.
• W1974865240 hasConcept C3020135677 @default.
• W1974865240 hasConcept C54355233 @default.
• W1974865240 hasConcept C71315377 @default.
• W1974865240 hasConcept C71924100 @default.
• W1974865240 hasConcept C86803240 @default.
• W1974865240 hasConceptScore W1974865240C126322002 @default.
• W1974865240 hasConceptScore W1974865240C134018914 @default.
• W1974865240 hasConceptScore W1974865240C142724271 @default.
• W1974865240 hasConceptScore W1974865240C207001950 @default.
• W1974865240 hasConceptScore W1974865240C2777702977 @default.
• W1974865240 hasConceptScore W1974865240C2778324911 @default.
• W1974865240 hasConceptScore W1974865240C2780394083 @default.
• W1974865240 hasConceptScore W1974865240C2780536345 @default.
• W1974865240 hasConceptScore W1974865240C3020135677 @default.
• W1974865240 hasConceptScore W1974865240C54355233 @default.
• W1974865240 hasConceptScore W1974865240C71315377 @default.
• W1974865240 hasConceptScore W1974865240C71924100 @default.
• W1974865240 hasConceptScore W1974865240C86803240 @default.
• W1974865240 hasFunder F4320320879 @default.
• W1974865240 hasIssue "5" @default.
• W1974865240 hasLocation W19748652401 @default.

• next