Diameter Of Secondary Follicles Research Articles

Influence of Diabetes on Morphometric Index of Ovarian Follicles in Streptozocin-Induced Rats

The study aimed to determine the effect of diabetes on follicle development by measuring the diameters of rats' ovarian follicles. The structure and function of many organs change by being affected by diabetes. The ovary is an essential organ of the reproductive system affected by diabetes. The size of the ovarian follicles and corpus luteum can also be affected by diabetes. For this reason, body mass, blood glucose level, and rat ovarian follicles, and corpus luteum diameters were measured in this study. Ten healthy female rats were kept as the control group. In the other rat group, experimental diabetes was induced with streptozotocin (STZ)(60 mg/kg). The rats in both groups were killed after 30 days and their ovaries removed. 5-6 µm sections were made using paraffin embedding techniques and stained with hematoxylin-eosin. On the fifth day of STZ administration to rats, the mass loss of rats was 10%, and the diabetogenic index was 330%. Compared with the control group, diameters of diabetic rats significantly decreased the diameter of the primordial, primary and Graafian follicles, and corpus luteum. The decrease in the diameter of secondary follicles of diabetic rats was not significant. The percentage shrinkage index was the highest in the corpus luteum with 37%. As a result, it can be said that diabetes influences the size of the ovarian follicles and especially the corpus luteum, thereby negatively affecting the ability to improve oocyte quality. Diabetes-related follicle diameter may shrink and cause infertility. It may be essential to measure the diameters of the follicles in vitro fertilization studies in patients with diabetes.

Black coffee mitigates diethyl phthalate disrupted folliculogenesis, reduced gonadotropins, and ovarian lesions in female albino mice.

Phthalates are multifunctional compounds with extensive applications and emerging environmental pollutants. Due to their ubiquity in the environment and unavoidable exposure to humans, concerns have been voiced about public health dangers. This study was aimed to explore the diethyl phthalate (DEP) toxicity and the potential protective effect of black coffee in female Swiss albino mice. Four-week-old mice, weighing 12 ± 1g were segregated into five groups (n = 10), designated as G-I (without any treatment), G-II (treated with corn oil), G-III (exposed to 1.5mg/g body wt. (B.W.) DEP), G-IV (received 2μg/g B.W coffee), and G-V (co-administrated with 1.5mg/g DEP and 2μg/g B.W coffee). Before dose administration, the coffee extract was assessed for its antioxidant potential through FRAP, TPC, and GC-MS analyses. Respective phthalates/coffee doses were administrated orally, once a day for 8weeks consecutively starting from the prepubescent stage. After 56days, mice were acclimated for 4days then dissected. Morphological assessments showed an irregular shape of the ovaries in DEP-treated mice as compared to the control. The average bodyweight of DEP-intoxicated mice (p ≤ 0.05) increased notably against control, while DEP plus coffee group showed a regular gain in the average weight of mice. The gonado-somatic index showed non-significant variations among all groups. Micrometric studies showed that the diameter of secondary follicles (115µm) in the ovaries of DEP-exposed mice (p ≤ 0.001) decreased significantly as compared to control (204µm); conversely, follicular diameter in the coffee control group (248) increased significantly. Serum FSH and LH levels were significantly increased in DEP-exposed mice with a noteworthy decrease in estrogen level while hormonal levels of all other groups were comparable to control. Histological sections of DEP-exposed mice ovaries showed anatomical disruptions contrary to other groups, which were comparable with control. Antioxidant potential was checked in ovaries homogenates; FRAP values showed a notable decrease in DEP group in comparison with the control group, in contrast to G-V, when DEP was co-administrated with coffee. This study concluded that black coffee has protective effect, against DEP-instigated reproductive toxicity in Swiss albino female mice.

EFFECTS OF ALOE VERA GEL ON MONOSODIUM GLUTAMATE INDUCED OVARIAN TOXICITY IN RATS

ABSTRACT: 
 Aim and Objective: Monosodium glutamate (MSG) is commonly used as a flavor enhancer. The aim of the study was to evaluate the possible protective effect of aloe vera gel on reproductive toxicity induced by MSG in female albino rats. Study design: Experimental study. Place of study: Anatomy department of Shaikh Zayed Postgraduate Medical Institute, Lahore.
 Material and Method: 36 female Wistar albino rats were randomly divided into three groups. Group A was the control group, it was given distilled water. The experimental group B was treated with 200 mg/kg b.w of MSG dissolved in 1.5 ml of distilled water, for 14 days by gastric intubation. While the experimental group C was provided with 400 mg/kg b.w of aloe vera gel extract, along with 200 mg/kg body weight of MSG dissolved in 1.5 ml of distilled water by gastric intubation for 14 consecutive days at an interval of 24 hours. The rats were dissected on the 15th day and their ovaries were removed. Light microscopic examination was done to study the histological changes.
 Results: The ovaries demonstrated deformed secondary and graafian follicles with vacuolization and there was a significant decrease in the diameters of secondary follicles (p value= 0.018) in group B. Histopathological changes induced by MSG in the ovaries were improved by co-administration of aloe vera gel in group C (p value=0.003). Conclusion: Aloe vera gel when consumed with MSG minimizes its harmful effects.

Immunohistochemical, morphological and histometrical analyses of follicular development in Astyanax bimaculatus (Teleostei: Characidae) exposed to an organochlorine insecticide

Thiodan® is an organochlorine insecticide used in agriculture that can reach aquatic ecosystems where it can affect fish reproduction. This research aimed to evaluate follicular development and the expression of integrin β1, collagen type IV and caspase 3 by morphological, histometrical and immunohistochemical analyses of Astyanax bimaculatus exposed to Thiodan®. Treatments included three sublethal concentrations of Thiodan® (1.15, 2.30, and 5.60μgL−1) for 96h and a control without the insecticide. Mature females with ovaries in advanced follicular development were chosen for study (average weight: 11.52 ± 2.0g; average total length: 9.12 ± 0.64cm). The follicles of A. bimaculatus exhibited normal morphology, while the diameters of secondary follicles showed an increase (P< 0.05) in all concentrations when compared to the control group; a characteristic of follicles undergoing the initial stages of intoxication. Immunohistochemical analysis revealed secondary follicles with greater expression of integrin β1 and collagen type IV in cytoplasm of follicular cells than in the primary follicles in all treatments and in the control. Immunolocalization of caspase 3 was detected in follicular cells during secondary development in all tested concentrations of Thiodan® and the control. These analyses demonstrate positive immunolocalization throughout the course of follicular development, even in fish exposed to varying concentrations of Thiodan® for 96h, demonstrating that follicular cells retain their physiological integrity.

Candidate genes for the development of hair follicles in Hu sheep.

The aim of this study was to detect candidate genes for the development of hair follicles in the Hu sheep breed. Seven genes have been detected in large, medium, and small wave follicles of Hu sheep using gene chip technology. The histological features of the follicles of newborn Hu-lambs were combined with fluorescence quantitative PCR technology to detect the correlation between the expression of the seven genes and hair follicle development. Among the genes studied, matrix metalloproteinase 2 (MMP2), bone morphogenetic protein-7 (BMP7), and sideroflexin 1 (SFXN1) showed a significantly different pattern of expression in large, medium, and small wave follicles (P < 0.05). The expression of MMP2 had a significant positive correlation with secondary follicles in large waves (P < 0.05), while the expression of BMP7 had a significant correlation with primary follicle diameter in small wave follicles, and a highly significant positive correlation with the number of secondary follicles in the small waves (P < 0.01). The expression of SFXN1 was significantly and positively correlated with the diameters of small wave primary follicles; it also showed a highly significant positive correlation with secondary follicle diameters. Although other genes are associated with hair follicles, their expression in large, medium, and small wave follicles was not significant. We propose that BMP7, MMP2, and SFXN1 genes could be important candidate genes for use in breeding Hu lambs with early coat development.

Effects of insecticide Thiodan®on the morphology and quantification of ovarian follicles in lambarisAstyanax bimaculatus(Linnaeus, 1758) in different treatments

Thiodan® is an insecticide which can cause morphological changes in fish tissues, dependent on the concentration. To investigate possible morphological changes, this study evaluated the number and diameter of ovarian follicles in lambaris Astyanax bimaculatus exposed to Thiodan® for 96 h in different treatments at three sub-lethal concentrations: 1.15, 2.3 and 5.6 μg L−1 and a control group without Thiodan®. Females exposed to Thiodan® were divided into four treatments: (i) without acclimation and without food, (ii) without acclimation and with food, (iii) with acclimation for 10 days and without food; (iv) with acclimation for 10 days and with food. In this study, it was noted that the action of Thiodan® at sub-lethal concentrations in a static system did not affect the morphology of follicular development. However, the treatments with acclimation showed lower numbers of primary and secondary follicles when compared to fish from treatments without acclimation. For the groups exposed to Thiodan®, a greater number of atretic follicles were observed compared to the control groups. The follicular diameter of secondary follicles in fish exposed to Thiodan® in the treatment with acclimation and with food was highest in the control group (P < 0.05). Furthermore, the mean gonadosomatic (GSI) and hepatosomatic (HSI) indices of treatments with acclimation were lower than those for treatments without acclimation. These data suggest that the fish may be being affected by the pesticide to which the fish were submitted, and may cause impairment to the follicular development.

Levels of BMP-6 mRNA in goat ovarian follicles and in vitro effects of BMP-6 on secondary follicle development

Expression of BMP-6 mRNA was quantified by real-time polymerase chain reaction (PCR) and the BMP-6 protein was demonstrated by immunohistochemistry in the primordial, primary, secondary, small and large antral follicles of goat. Furthermore, the influence of BMP-6 on increase in diameter, antrum formation and expression of BMP-6 and FSH-R in in vitro cultured secondary follicles was studied. Therefore, goat primordial, primary and secondary follicles, as well as small and large antral follicles were obtained and the mRNA levels of BMP-6 were quantified by PCR in real time. Expression of BMP-6 protein in goat follicles was demonstrated by immunohistochemistry. The influence of BMP-6 in the presence or absence of follicle-stimulating hormone (FSH) on both the development of secondary follicles and the expression of mRNA for BMP-6 and FSH-R was evaluated after 6 days of culture. Furthermore, the follicular diameter and the formation of the antrum were evaluated before and after 6 days of culture and compared by Kruskal-Wallis and chi-squared tests (P < 0.05), respectively. The results show that the level of mRNA for BMP-6 in primary and secondary follicles was significantly higher than in the primordial follicles (P < 0.05). Similar levels of BMP-6 mRNA were observed in cumulus-oocyte complexes and mural granulosa/theca cells from small and large antral follicles, respectively. BMP-6 protein was expressed in oocytes of all categories of follicles and in granulosa cells from secondary follicles onwards. Addition of BMP-6 to the culture medium increased the diameter of secondary follicles mainly by antrum formation after 6 days' culture, in the presence or absence of FSH (P < 0.05). Furthermore, addition of FSH resulted in increased levels of BMP-6 mRNA in these follicles (P < 0.05). Simultaneous administration of FSH and BMP-6 enhanced the levels of FSH receptor (FSH-R) mRNA (P < 0.05). It is concluded that BMP-6 mRNA is increased during transition from primordial to primary/secondary follicles in the goat ovaries and that BMP-6 enhances the growth of cultured secondary follicles.

In vitro culture of sheep lamb ovarian cortical tissue in a sequential culture medium

This study evaluates the effect of a sequential culture system on the follicular development of sheep lamb ovaries, aiming to establish an available in vitro culture system for ovarian culture. Lamb ovarian cortical fragments were cultured on a steel mesh with a nitrocellulose membrane pre-coated by type 1 collagen. Several culture media were used for the determinations, specifically, a control medium (alpha-MEM), a constant medium (control medium supplemented with 75 ng/mL human recombinant EGF, 200 mIU/mL sheep FSH, 100 ng/mL human recombinant GDF-9, and 100 ng/mL human recombinant bFGF), and a sequential medium (control medium supplemented with sequential growth factors added on different days). Ovarian tissues, both fresh and cultured, were processed for histological and apoptotic assays, while spent culture media were processed for hormone assays. It was found that the growth of lamb primordial follicles can be initiated during culture in vitro. Compared to the control medium, sequential culture medium significantly increased the percentage of secondary follicles in cultures, while the follicle and oocyte diameters of primary and secondary follicles were also observed to increase in this medium. The constant medium was found to increase the number and diameter of secondary follicles only 18 days after culture. After this same period of time, some normal antral follicles were found in the sequential medium, while a few abnormal antral-like follicles were found in the control medium. Moreover, sequential medium appeared to significantly increase estradiol and inhibin production, especially 10-18 days after culture. The highest percentage of normal follicles and the lowest apoptotic cell rates were observed in the sequential medium, suggesting that a sequential addition style of culture can improve follicle and tissue viability. The sequential addition of FSH, EGF, GDF-9,and bFGF can stimulate primordial follicle transmittal into the later development stages, even as far as the antral stage, improve the survival rate of follicles, and maintain follicular viability.

Investigation of the Effects of Aloe barbadensis on Rat Ovaries: A Preliminary Study

Effects of Aloe barbadensis, a type of Aloe vera, on ovaries were investigated during pregnancy. A. vera gel, a commercial and nontherapeutic form of A. barbadensis, was used for this purpose. Three groups (one control and two test groups) of female Wistar albino rats with no prior births were studied. Group I was administered 25 mg (140 mg/kg) of A. barbadensis/day both orally and through gavage. A. barbadensis was contained in capsules with 500 mg of soybean oil; therefore Group II was administered 500 mg of soybean oil. Group III was the control group. All three groups were given normal food and water ad libitum. The substance had been administered for 20 days until birth. Ovaries were examined histologically. Vascular increase and the hyperemic form of ovary in the group administered A. barbadensis were notable. Decrease in primary follicle numbers, increase in secondary follicle numbers, and diminishment of secondary follicle diameters occurred in ovaries. The histological changes imply an angiogenesis effect of A. barbadensis and an effect like that of follicle-stimulating hormone in ovaries.

Bovine oocytes in secondary follicles grow and acquire meiotic competence in severe combined immunodeficient mice.

Cortical tissues containing only primordial and primary follicles, or secondary follicles 140-190 microm in diameter, were collected from bovine ovaries and xenografted under the kidney capsules of female severe combined immunodeficient (SCID) mice. Histological examination revealed that all grafts were well vascularised and contained surviving follicles at 4 or 6 weeks after grafting. Primordial and primary follicles survived but did not develop beyond the one-layer stage. Secondary follicles, on the other hand, had formed antra at 4 weeks after grafting. The mean diameter of secondary follicles, which was 165.2 +/- 17.0 microm (n = 42) before grafting, had developed to 442.9 +/- 77.9 microm (n = 37) and 592.9 +/- 116.0 microm (n = 45) in diameter at 4 and 6 weeks after grafting, respectively. The mean diameter of oocytes, which was 55.1 +/- 4.9 microm (n = 42) before grafting, also increased significantly (4 weeks: 105.6 +/- 6.3 microm; 6 weeks: 122.2 +/- 2.6 microm; p < 0.05). Oocytes were recovered from follicles that had developed to more than 400 microm in diameter after 6 weeks, and were subjected to subsequent mature culture. Of these oocytes, 34% (11/32) resumed meiosis and 6% (2/32) matured to the second metaphase. Follicular fluid in bovine antral follicles developed in SCID mice had the 69 kDa protein, which was detected by anti-mouse albumin antibody but not by anti-bovine albumin antibody in immunoblotting analysis. These results demonstrated that bovine secondary follicles develop to the antral stage in SCID mice, and that the oocytes in the follicles acquire the meiotic competence.

Skin histology and birthcoat fibre-follicle relationships in Merino and ⅝ Merino lambs

SUMMARYSkin biopsy samples from imported Merino and ⅝ Merino/⅜ Barki were examined from birth to the age of 1 year. Birthcoat fibre type arrays and fibre-follicle relationships were also studied.Merino lambs had a greater S/P ratio than ⅝ Merino at all ages; values at 1 year of age were 9·98 and 6·87 respectively.The ⅝ Merino lambs showed a wider range of follicle diameters than Merinos from birth to 1 year of age.The ratio of primary to secondary follicle diameters showed higher values in ⅝ Merino than in Merinos. Merino gave lower values of follicle wall thickness than ⅝ Merino.In Merino birthcoat samples, only one array was plain, representing 4%, whereas the rest were saddle. In ⅝ Merino the majority of fibre type arrays were plain, representing 75 %, whereas the rest were saddle and plateau.In Merino, because of the absence of halo hairs and super sickles, only two samples had few super sickle B; primary central follicles were occupied by sickle fibres, medium with large sickle ends as well as curly tips, and primary lateral follicles had mostly medium and fine curly tips, only four samples had sickle fibres, and secondary follicles produced medium and fine curly tips and histerotrichs.In ⅝ Merino primary central follicles produced halo hairs, super sickle A, A', B and occasionally sickle fibres, primary lateral follicles grew super sickle A, A', B, sickle fibres and curly tip fibres, and secondary follicles had medium and fine curly tips and histerotrichs.

Wool follicle characteristics in the Awassi fat-tailed sheep.

In Awassi sheep, the total number of wool follicles per square millimetre was affected significantly by age, sex, and type of birth. The density of the primary follicles was only significantly affected by age and type of birth. The secondary follicles to primary follicles (S/P) ratio values were 2.0 at birth, 4.0 at 2 months and 4.2 at 6 months of age, in Awassi sheep. The males exceeded the females, in most ages of the experiment. The single lambs surpassed the twins in the S/P ratio values. The effect of age and type of birth on the S/P ratio was highly significant. The external diameter of wool follicles increased with higher values at 2 months of age than afterwards, till 6 months of age. The contrary was the case in the internal diameters of wool follicles. The increase was higher in the primary than in the secondary follicles, during the period of the study. The males surpassed, in general, the females in the values of the external and internal diameters of the primary and secondary follicles. Single lambs showed higher values than twins in the external and internal diameters of primary and secondary follicles in most ages of the experiment. Age and type of birth affected significantly the diameters of follicles, while sex effect was only highly significant on the external diameter of the secondary follicles. The length of the primary and secondary follicles and the thickness of skin increased significantly, as the animals grew older. Sex showed no significant effect on the length of follicles, while it affected significantly the thickness of the skin. Single lambs showed higher significant values than twins, in length of follicles and skin thickness. The bulbs of each of the primary and secondary follicles increased significantly, as the animals grew older. Sex had no significant effect on the bulb diameters of the two types of follicles. The single lambs surpassed the twins in the values for follicle bulb diameter, in most ages of the experiment. The effect of the type of birth on the bulb diameter of the two types of follicles was highly significant.

Studies of face cover in the New Zealand Romney Marsh sheep. II. The measurement of face cover and of factors associated with differences in face grades

Face cover was measured as poll staple length and the length of the nose free of visible wool growth. Forty open-faced and 80 woolly-faced ewes were measured every June for five years. While differences in the measurements occurred between the grades, analyses showed that neither measurement, nor a combination of them, discriminated satisfactorily between grades over the full range of face cover. Within a face-cover grade, poll staple length was positively correlated with the midside staple length. Between the grades the relationship was a negative one. Follicle characteristics were measured in facial skin from seven open-faced and 10 woolly-faced sheep from the same flock. Open-faced sheep had the larger primary follicles but smaller secondary follicle diameters. Thus open-faced sheep had a dp/ds ratio of 2.3 and woolly-faced sheep one of 1.4. Further skin samples from six rams and seven wethers at 1 year of age showed that castration had reduced primary follicle diameter (57 and 41 µ) without changing the face cover. Also ewes and ovariectomized ewes showed a difference in SIP ratio on the face (1.9 and 2.5) with no difference in face cover. It was concluded that primary follicle size was not an immediate determinant of face cover but that factors external to the facial follicles were more likely to be of importance.