Microsurgical treatment of distal anterior cerebral artery aneurysms: A 25 year institutional experience.

INTRODUCTION: We present a retrospective study of distal anterior cerebral artery (DACA) aneurysms operated at our institute. MATERIAL AND METHODS: From 1988 to 2013, a total of 132 DACA aneurysms were managed surgically. The retrospective data of these patients was collected and analysed. RESULTS: They comprised 6.2% of all 2137 aneurysms operated during the same period. The mean age at presentation was 48.3 years (range: 24-77 years) and the male: female ratio was 1:2. The notable symptom in 114 (86.4%) patients was headache, in 94 (71.2%) was loss of consciousness, in 25 (18.9%) was seizures, and in 17 (12.9%) was limb weakness. There were 99 (75%) patients in World Federation of Neurosurgical Societies (WFNS) grade <3 at admission. On plain computed tomography (CT) scan, intracerebral hemorrhage was seen in 49 (37.1%) and intraventricular hemorrhage in 23 (17.4%) patients. The most common site of aneurysm was the pericallosal (A3 segment) artery noted in 65 (49.2%) patients. Multiple aneurysms were observed in 16 (12.1%) cases. The mean timing of surgery from the first ictus was 17.8 days. The delay was due to a late referral, poor general condition, recurrent bleeding, and worse WFNS grade at presentation. Aneurysms were operated through the following approaches: parasaggital craniotomy in 61 (46.2%), bifrontal craniotomy in 64 (48.5%), and frontotemporal craniotomy in 7 (5.3%). Aneurysms could be successfully clipped in 93.2% of the cases. Postoperative mortality was 8 (6%). The follow-up data was available for 83 (62.9%) patients. The mean duration of follow-up was 15.1 months (1-70 months), with more than 41 (31%) patients having a follow-up of more than 6 months. A favorable outcome (good recovery or moderate disability) was seen in 71 (85.5%) cases. CONCLUSIONS: A meticulous attention to the surgical technique provides gratifying results in DACA aneurysms even in patients who present initially in a compromised clinical status.

Chicken sperm transcriptome profiling by microarray analysis.

It has been confirmed that mammalian sperm contain thousands of functional RNAs, and some of them have vital roles in fertilization and early embryonic development. Therefore, we attempted to characterize transcriptome of the sperm of fertile chickens using microarray analysis. Spermatozoal RNA was pooled from 10 fertile males and used for RNA preparation. Prior to performing the microarray, RNA quality was assessed using a bioanalyzer, and gDNA and somatic cell RNA contamination was assessed by CD4 and PTPRC gene amplification. The chicken sperm transcriptome was cross-examined by analysing sperm and testes RNA on a 4 × 44K chicken array, and results were verified by RT-PCR. Microarray analysis identified 21,639 predominantly nuclear-encoded transcripts in chicken sperm. The majority (66.55%) of the sperm transcripts were shared with the testes, while surprisingly, 33.45% transcripts were detected (raw signal intensity greater than 50) only in the sperm and not in the testes. The greatest proportion of up-regulated transcripts were responsible for signal transduction (63.20%) followed by embryonic development (56.76%) and cell structure (56.25%). Of the 20 most abundant transcripts, 18 remain uncharacterized, whereas the least abundant genes were mostly associated with the ribosome. These findings lay a foundation for more detailed investigations on sperm RNAs in chickens to identify sperm-based biomarkers for fertility.

Development of a new method for sperm RNA purification in the chicken.

Currently RNA transcripts are being used as male fertility biomarker for many mammalian species, but research work on chicken is at halt because classical RNA isolation methods are not effective for chicken spermatozoa. Hence, attempts have been made to optimize RNA isolation protocol from chicken sperm by using different methods, and to confirm the presence of sperm-specific transcripts of PRM and PLCZ1. Semen samples were centrifuged at low speed for removing debris like uric acid. Further, 1mL diluted semen was gently placed over 40% PureSperm or 45%/90% Percoll, and centrifuged to remove somatic cells and immature diploid spermatocytes. RNA was isolated from sperm by using RNAzol or TRIzol reagent or RNeasy Micro kit with certain modification, and RNA quantity and quality were evaluated. RNA isolated by using RNAzol or RNeasy Micro Kit yielded good quantity and quality of RNA for downstream applications compared to TRIzol. 40% PureSperm was found effective in removing somatic cells. RT-PCR results showed that sperm RNA samples were negative for CD4 and PTPRC. All the sperm RNA samples were positive for PRM and PLCZ1, markers of sperm RNA.

Sexual maturation, serum steroid concentrations, and mRNA expression of IGF-1, luteinizing and progesterone hormone receptors and survivin gene in Japanese quail hens.

In avian species, sexual maturation represents the evidence of start laying, which is a consequence of the development of ovarian follicles. These follicles are the functional reproductive unit whose maturation and viability critically depends on endocrine, paracrine, and autocrine factors beyond the signals from the central nervous system. The present study was undertaken to investigate the correlation of sexual maturity with tissue growth, mRNA expression of certain genes, and serum steroid concentrations in Japanese quail hens. To carry out the present study, a total of forty Japanese quail hens (5 weeks) were housed individually under uniform husbandry condition with ad libitum quail layer ration and water at 14-hour photo schedule. On sixth week onwards, four birds were sacrificed at each time on 1, 3, 7, 10, 13, 16, 19, 22, 25, and 28 days. Serum was extracted aseptically to analyze the gonadal steroid hormones (estrogen and progesterone) and corticosterone to investigate the liaison with sexual maturation of the species. Expression analyses of four genes i.e., insulin-like growth factor-1, luteinizing hormone receptor, progesterone receptor, and survivin were carried out in the three largest ovarian yellow follicles. A significant (P < 0.05) increase in body weight gain and oviduct weight was recorded during the phase of sexual maturation. Smaller follicles revealed higher insulin-like growth factor-1 and survivin gene expression, whereas the reverse result was manifested in both the luteinizing and progesterone hormone receptors. In biochemical study, the gonadal steroids (estrogen and progesterone) were recorded higher at the first half of the experiment when a gradual decrease in corticosterone concentration was confirmed from the very beginning of this study. This result substantiated that sexual maturation in Japanese quail may be completed by the time of 8 weeks after its birth in support of the analyzed information studied in the current investigation.

Effect of dietary supplementation of vitamin E on production performance and some biochemical characteristics of cloacal foam in male Japanese quail.

This experiment was conducted to investigate the effects of increasing the level of dietary supplementation of vitamin E (VE) on production performance and biochemical characteristics of cloacal foam in male Japanese quail (Coturnix coturnix japonica). A total of 225 male Japanese quail chicks (day old) were randomly distributed to three dietary treatments for a period of 30 weeks. Each treatment comprised of three replicates, each containing 25 chicks. The basal diet (T1) contained 12.30IUVEkg(-1) and the two experimental diets were supplemented with 150 and 300IUVEkg(-1) (diets T2 and T3, respectively). dl-α-Tocopherol acetate was used as the source of VE. All chicks were provided feed and water ad libitum. Mean body weights, feed intake, feed conversion ratio (FCR) and mortality of the birds in the different treatment groups showed no significant differences (P>0.05), whereas a significant (P<0.05) increase (29.81 and 50.83%) in average foam weight was evident in the VE-treated groups (T2) compared with control (T1) and T3 groups. The biochemical characteristics of foam, in terms of quantities of protein and nitric oxide (NO), did not differ significantly (P>0.05), whereas the quantities of glucose (60.01%) and acid phosphatase (ACP, 32.46%) were significantly (P<0.05) higher in the T3 group. By contrast, the quantities of alkaline phosphatase (ALP), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were significantly (P<0.05) lower (48.84%, 10.38% and 22.08%, respectively) in the T3 group and higher in the T1 (control) and T2 groups. From this study, it can be concluded that dietary supplementation of VE to the basal diet has no effect on the production performance but supplementation of a higher level of VE (300IUkg(-1)diet) improved the biochemical characteristics of the foam and moderate levels of VE (150IUkg(-1)diet) improved the foam production of male Japanese quail.

In vitro initiation of the acrosome reaction in the emu (Dromaius novaehollandiae).

1. An assessment of the efficiency of the acrosome reaction (AR) provides an important predictor of the fertilizing potential of semen and for diagnosis of the causes of infertility. A standardized protocol was therefore developed for initiation of the acrosome reaction in emu spermatozoa in vitro, and the role of CaCl2 or perivitelline membrane (PVM) proteins in determining the outcome of the reaction was investigated. 2. The acrosome reaction (assessed by FITC-PNA) was successfully induced in live spermatozoa by incubation for 2 min in NaCl-TES medium supplemented with 5 mM CaCl2. The maximum response was 32% live acrosome-reacted spermatozoa (LAR) achieved after 10 min incubation. 3. Compared to the outcome with 5 mM CaCl2 or PVM protein alone, the response was significantly better with a combination of PVM protein and CaCl2. 4. A significant variation in the percentage of LAR spermatozoa among individual males was observed. No treatment affected the percentage of dead acrosome-reacted spermatozoa. 5. The results emphasize the important role played by both PVM proteins and Ca(2+) in the in vitro initiation of the acrosome reaction.

Ovarian morphology and internal vis-à-vis non internal laying in relation to triacylglycerol, hormones and their receptors concentration around the age of sexual maturity in broiler breeder hens.

1. Ovarian morphology, serum hormone concentrations of 17-ß-estradiol, progesterone, testosterone, tri-iodothyronine (T3), thyroxine (T4) and triacylglycerol (TAG) were investigated at 23 and 26 weeks of age in broiler breeder hens provided with ad libitum access to feed. Progesterone, oestrogen-ß, thyroid-α and -ß receptor mRNAs were also quantified in the infundibulum at the same ages. 2. A large variation in the ovarian morphology was observed at 23 weeks of age including hens with undeveloped ovaries, non-laying hens with post ovulatory follicles (POF) and a predominance of non-laying hens without a POF. 3. Serum concentrations of triglyceride, 17-ß-estradiol and progesterone at 23 weeks of age were lower in hens with an undeveloped ovary compared with other groups of hens, whereas testosterone, triiodothyronine and thyroxin were higher. 4. At 26 weeks of age, the average number of hierarchical yellow follicles in normal layers was 7.64 ± 0·41 whereas in internal layers, the follicular numbers were significantly greater at 8.66 ± 0·53. The higher follicular numbers in internal layers were associated with higher serum triglyceride and progesterone concentrations. 5. Oestrogen receptor-ß and thyroid receptor-ß mRNA was up regulated in the infundibulum of internal layers compared with normal laying hens at 26 weeks of age.

Expression profile of luteinizing hormone receptor gene in hierarchal follicles and regressing oviduct tissues of White Leghorn hens during moulting.

In the present study, the expression profile of luteinizing hormone receptor (LHR) was investigated in the ovary, magnum and uterus and in hierarchcal follicles (F-1, F-2, F-3 and F-4) of hens subjected to moulting to establish their involvement in moulting and presence in non-gonadal tissues. Fifty-two layers (72 weeks) were subjected to moult for a period of 14 days. Four birds were sacrificed each time on 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12 and 14 days of moulting, and samples (ovary, magnum, uterus and hierarchal follicles) were collected aseptically for the quantitative study by real-time polymerase chain reaction. The ovary, isthmus, uterus and magnum weight reduced significantly during induced moulting. From the 4 DOM, this reduction was drastic and reached approximately 80% of original weight in the case of ovary, isthmus and magnum and approximately 65% of original weight in the case of uterus on 14 DOM. Ovarian yellow follicles decreased gradually from 1 DOM to 4 DOM, after that no normal yellow follicle was observed in moulted bird. The number of atretic follicles increased gradually during the course of induced moulting, reaching the peak at 5 DOM. The LHR mRNA was detected in non-gonadal tissues like magnum and uterus. The LHR expression was significantly (p < 0.05) down regulated in ovary, magnum and uterus throughout the treatment. These results indicated that LHR may have a role in reproductive tissue regression during moulting.

Expression of leptin and its receptor in corpus luteum during estrous cycle in buffalo (Bubalus bubalis).

Leptin is supposed to play a crucial role in ovarian luteal dynamics. The present study was aimed to investigate the importance of leptin and its receptors in buffalo corpus luteum (CL) obtained from different stages of the estrous cycle. Real-time RT-PCR (qPCR), western blot and immunohistochemistry techniques were applied to investigate mRNA expression, protein expression and localization of examined factors. Additionally to assess the contribution of leptin in progesterone production the expression profiles of StAR, P450scc and HSD were also investigated. In general, we demonstrated presence of leptin and its receptors in buffalo CL during the estrous cycle. The mRNA levels of leptin and its receptors were significantly up regulated in (P<0.05) in all the stages and highest levels were observed in mid and late luteal stages consistent with in vivo luteinization of buffalo CL and declined coincidental to luteal regression. The expression of StAR, P450scc and HSD factors maintained low in early luteal phase, after that level of expression increased steadily to show a significant rise (P<0.05) in mid luteal phase followed by gradual decline in late luteal phase and regressed CL and this correlates well with the Ob and ObR receptor activity, verifying their key role in progesterone and other steroids production in functional CL. As revealed by immunohistochemistry, leptin protein was localized predominantly in large luteal cells however leptin receptor (Ob-R) was localized in large luteal cells as well as in endothelial cells. It can be concluded from our study that leptin via its autocrine/paracrine effects play a significant role in promoting angiogenesis, steroidogenesis and also acts as key survival factor in bubaline CL.

The role of the male cloacal gland in reproductive success in Japanese quail (Coturnix japonica).

The adult male Japanese quail has a well developed cloacal gland that produces meringue-like white foam. The physiological significance of the cloacal gland and its foam is still unclear. Therefore, we conducted two experiments to establish the functional role of the cloacal gland and its foam in natural mating and oviducal sperm transport. In the first experiment, artificial insemination of equal numbers of spermatozoa diluted in foam extract and normal saline once in a week were used to determine the role of foam in sperm transport in the female oviduct. After artificial insemination, eggs were collected to measure fertility, the duration of the fertile period, sperm holes and attached spermatozoa in the perivitelline membrane. Higher (P<0.05) fertility and greater duration of the fertile period were observed when semen was inseminated along with foam extract compared with normal saline. Further, the sperm holes and trapped spermatozoa in the perivitelline membrane were also higher (P<0.05) in the presence of foam extract. In the second experiment, two males with bigger and smaller cloacal gland areas were allowed to mate with a female. The mating attempts of males with larger cloacal gland were more successful (P<0.05) than males with smaller cloacal glands. Our results indicated that cloacal foam improves sperm transport in the female oviduct and that males with larger cloacal gland areas are preferred during mating.

Effect of Dietary L-ascorbic Acid (L-AA) on Production Performance, Egg Quality Traits and Fertility in Japanese Quail (Coturnix japonica) at Low Ambient Temperature.

Environmental stress boosts the levels of stress hormones and accelerates energy expenditure which subsequently imbalance the body's homeostasis. L-ascorbic acid (L-AA) has been recognized to mitigate the negative impact of environmental stress on production performances in birds. The present investigation was carried out to elucidate the effect of different dietary levels of L-AA on production performance, egg quality traits and fertility in Japanese quail at low ambient temperature. Sixty matured females (15 wks) were equally divided into three groups (20/group) based on the different dietary levels of L-AA (0, 250 and 500 ppm) and coupled with an equal number of males (1:1) obtained from the same hatch. They were managed in uniform husbandry conditions without restriction of feed and water at 14 h photo-schedule. Except for feed efficiency, body weight change, feed consumption and hen-day egg production were recorded highest in 500 ppm L-AA supplemented groups. Among the all egg quality traits studied, only specific gravity, shell weight and thickness differed significantly (p<0.05) in the present study. Fertility was improved significantly (p<0.01) to a dose dependent manner of L-AA. The findings of the present study concluded that dietary L-AA can be a caring management practice at least in part to alleviate the adverse effect of cold induced stress on production performance in Japanese quail.

Influence of chicken native breeds on some physical and biochemical characteristics and short-term storage of semen.

1. The major objective of this study was to examine the influence of 24-h storage of semen at low temperature on semen characteristics and fertilising ability of spermatozoa in two native breeds (Kadaknath-KN, Aseel Peela-AP) and White Leghorn (WL) chicken. 2. Various physical and biochemical properties of freshly ejaculated semen of KN and AP were investigated. Fertility was examined in freshly-ejaculated as well as 24-h-stored (3°C) semen diluted (1:3) with Beltsville Poultry Semen Extender. 3. No significant difference was observed in sperm motility among the different breeds whereas live counts were higher in WL than the native breeds. Body weight, semen volume and sperm concentration were highest in AP, followed by KN and WL. A similar trend was observed in the percentage of dead and morphologically-abnormal spermatozoa. 4. The activity of acid and alkaline phosphatase in seminal plasma were higher in WL than KN, whereas the opposite trend was recorded for glutamic oxaloacetic and pyruvic transaminases. The cholesterol content of semen was highest in AP, followed by KN and WL. Cholesterol was much lower in seminal plasma compared with whole semen but there were no differences between breeds. Mean values of the methylene blue reduction time test were higher in WL than in the native breeds. 5. Fertility and hatchability, using freshly-diluted semen, were poorer in the native breeds than in WL. The pattern of fertility deteriorated further, especially in native fowls, when the birds were inseminated with 24-h-stored semen. 6. In conclusion, variation in physical and biochemical characteristics of semen in native breeds compared to WL correlated with poor fertility after short-term storage of semen.

Cloacal gland foam enhances motility and disaggregation of spermatozoa in Japanese quail (Coturnix japonica).

The adult male Japanese quail produces white foam from the cloacal gland, which is transferred to the female proctodeum during natural mating. The physiological role of foam on quail spermatozoa is still unclear. Therefore, attempts have been made to understand the effect of cloacal foam on motility and metabolism of quail spermatozoa. The profile of various biochemical constitutes in the foam extract was investigated. The addition of foam extract to neat semen completely disaggregated the clumps of spermatozoa leading to vigorous motility. The metabolic rate (MBRT) of the spermatozoa was significantly increased with the addition of foam extract. The foam extract was sub fractionated into seven different fractions by using the molecular cut off devices. Among all the seven sub-fractions from the foam extract, the addition of < 1 KDa sub-fraction contained lactate and has enhanced sperm motility and metabolism. Another fraction (3-10 KDa) has non-protein and non-heparin components which completely disaggregated the clumped quail spermatozoa. However, the remaining fractions did not show any effect on quail spermatozoa. It can be concluded from the present investigation that the lactate present in foam might be a fuel for sperm metabolism and motility. Furthermore, low molecular weight (3-10 KDa) components in the foam may responsible for sperm disaggregation.

Characterization of lactate dehydrogenase enzyme in seminal plasma of Japanese quail (Coturnix coturnix japonica).

Lactate dehydrogenase enzyme present in quail seminal plasma has been characterized. Polyacrylamide gel electrophoresis and subsequently with LDH specific staining of seminal plasma revealed a single isozyme in quail semen. Studies on substrate inhibition, pH for optimum activity and inhibitor (urea) indicated the isozyme present in the quail semen has catalytic properties like LDH-1 viz. H-type. Furthermore, unlike other mammalian species, electrophoretic and kinetic investigations did not support the existence of semen specific LDH-X isozyme in quail semen. The effect of exogenous lactate and pyruvate on sperm metabolic activity was also studied. The addition of 1 mM lactate or pyruvate to quail semen increased sperm metabolic activity. Our results suggested that both pyruvate and lactate could be used by quail spermatozoa to maintain their basic functions. Since the H-type isozyme is important for conversion of lactate to pyruvate under anaerobic conditions it was postulated that exogenous lactate being converted into pyruvate via LDH present in semen may be used by sperm mitochondria to generate ATP. During conversion of lactate to pyruvate NADH is being generated that may be useful for maintaining sperm mitochondrial membrane potential.

Effect of higher dietary vitamin E concentrations on physical and biochemical characteristics of semen in Kadaknath cockerels.

1. This experiment was to investigate the effects of increasing dietary vitamin E on physical and biochemical characteristics of semen in Indian reared Kadaknath (KN) cockerels. DL-alpha-Tocopherol acetate was used as the source of vitamin E. 2. A total of 135 one-day-old male KN chicks were randomly selected and divided into 9 groups with 15 chicks in each group (3 dietary treatments x 3 replicates). 3. The basal diet contained 15 IU (10 mg) vitamin E/kg and the two experimental diets were supplemented with 150 IU (100 mg) and 300 IU (200 mg) vitamin E/kg (diets T(2) and T(3), respectively). 4. Physical characteristics in terms of semen volume, sperm concentration, sperm motility and percentage live sperm did not differ significantly, whereas proportion of abnormal and dead spermatozoa were significantly lower and fertility higher in the T(2) group. 5. Biochemical characteristics in term of quantities of protein and nitric oxide (NO) did not differ significantly, whereas the quantity of glucose, acid phosphatase (ACP) and vitamin E were significantly higher in the T(2) group. 6. In contrast, the quantities of alkaline phosphatase (ALP), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were significantly lower in T(2) group and higher in the T(1) (control) group. 7. From this study it can be concluded that moderate supplementation of dietary vitamin E may be beneficial for physical and biochemical characteristics of semen in Indian reared KN cock.

Fabrication of cesium-137 brachytherapy sources using vitrification technology.

137Cs source in solid matrix encapsulated in stainless-steel at MBq (mCi) levels are widely used as brachytherapy sources for the treatment of carcinoma of cervix uteri. This article describes the large-scale preparation of such sources. The process of fabrication includes vitrification of 137Cs-sodium borosilicate glass, its transformation into spheres of 5-6 mm diameter, casting of glass spheres into a cylinder of 1.5 mm (varphi) x 80 mm (l) in a platinum mould, cutting of the moulds into 5-mm-long pieces, silver coating on the sources, and finally, encapsulation in stainless steel capsules. Development of safety precautions used to trap 137Cs escaping during borosilicate glass preparation is also described. The leach rates of the radioactive sources prepared by the above technology were within permissible limits, and the sources could be used for encapsulation in stainless steel capsules and supplied for brachytherapy applications. This development was aimed at promoting the potential utility of 137Cs-brachytherapy sources in the country and reducing the user's reliance on imported sources. Since its development, more than 1000 such sources have been made by using 4.66 TBq(126 Ci) of 137Cs.

Recovery and purification of 4.66 TBq(126Ci) of 137Cs from a 20-year-old spent sealed source.

An attempt to recover and purify 4.66 TBq(126Ci) of (137)Cs as solution from a 20-year-old doubly encapsulated sealed source containing (137)CsCl was made successfully. The primary capsule was first cut open to retrieve the secondary capsule. The top end of the secondary SS capsule was then cut to render it as an open cylinder and the (137)CsCl pellet inside was dissolved in small aliquots of water. Each aliquot of the (137)CsCl solution was dispensed into glass vials, sealed, assayed and stored. Barium formed during the radioactive decay of (137)Cs was removed by carbonate precipitation. Chloride was removed by heating with HNO(3) and evaporation. About 91.8% of the total activity could be recovered and used subsequently for making brachytherapy sources.

Teletherapy sources with imported and indigenous Co activity.

Board of Radiation and Isotope Technology, a unit of the Department of Atomic Energy, fabricates and supplies radioactive sources for medical, industrial, agriculture and research applications. High specific activity cobalt-60, required for teletherapy is normally imported. There was a proposal for manufacturing high specific activity sources indigenously. A study was carried out to observe the feasibility of mixing imported and indigenous cobalt-60 pellets to fabricate teletherapy source capsules. The specific activity of imported pellets is more than 300 Ci/g, whereas that of indigenous pellets obtained from Indian power reactors is 140 Ci/g. The radiation output from a capsule for different combinations of specific activity was evaluated. Losses due to self-absorption were accounted in the evaluations. In another study, the optimized lengths of the capsule for an output of 200 RMM and the additional activity to be added to compensate losses due to self-absorption were also estimated for different specific activity pellets. Sources fabricated on the basis of this study showed a good agreement with the estimations. Source capsules with a combination of different specific activities are yet to be fabricated.

Expression analysis of melatonin receptor subtypes in the ovary of domestic chicken.

Melatonin (N-acetyl-5-methoxytryptamine), an indole hormone, regulates various biological functions through three different receptor subtypes (Mel-1a, Mel-1b, and Mel-1c). However, the distribution of different melatonin receptor subtypes in chicken reproductive tissues was not known. In the present investigation, the partial sequences of ovarian melatonin receptor subtypes (Mel-1a, Mel-1b, and Mel-1c) were characterized. Further, the expression profile of melatonin receptor subtypes in the granulosa and theca layers of different preovulatory and postovulatory follicles (POF) were studied by semi-quantitative RT-PCR. The expression of all three subtypes of melatonin receptors were observed in the ovary of domestic chicken. Analysis of partial sequences of ovarian melatonin receptors revealed that the melatonin subtypes were identical to the brain receptors. In small white ovary follicles, we observed only the expression of mel-1b receptors, but not mel-1a or mel-1c receptors. In yellow follicles, all the three subtypes of receptors expression were noticed. Interestingly, we observed the expression of mel-1a receptor only in thecal layer, but not in granulosa layer. In contrast, mel-1b and -1c receptors were expressed in both granulosa and thecal layer. During the regression of POF, we observed significant upregulation of melatonin receptors (mel-1a and 1c) expression, that downregulated in the later stages of regression. We assume that the expression of melatonin receptors might have been influenced by the atresia or apoptosis of different follicular layers in POF. Our findings suggest that the differential distribution of melatonin receptor subtypes might have distinct downstream cellular functions in the ovarian tissues.

High doses of dietary zinc induce cytokines, chemokines, and apoptosis in reproductive tissues during regression.

In chickens, high levels of dietary zinc cause molting, and the reproductive system undergoes complete remodeling concomitant to feather replacement. In the present study, the expression profiles of cytokines and chemokines were investigated in the ovary and oviduct of control hens and of hens induced to molt by zinc feeding. The zinc-induced feed-intake suppression, the changes in corticosterone levels, the immune cell populations in the reproductive tract, and the apoptosis of reproductive tissues were analyzed. The expression of mRNAs for interleukin-6 (IL-6), interferon-gamma (IFN-gamma), the avian ortholog of mammalian IL-8 (chCXCLi2), and a chicken MIP-1beta-like chemokine (chCCLi2) in the ovary and of mRNAs for IL-1beta, IL-6, IFN-gamma, transforming growth factor-beta2, chCXCLi2, and chCCLi2 in the oviduct were upregulated significantly during zinc-induced molting. A simultaneous feed-intake reduction was observed with higher expression of cytokines and chemokines. The results of the present investigation also suggested that the upregulation of corticosterone was closely associated with the increased expression of cytokines and chemokines. An increase in apoptosis within reproductive tissue during tissue regression was also noted. We had previously observed the upregulation of these cytokines expression in an earlier study (molting by feed withdrawal). However, the pattern and the level of expression were different among these two methods. These findings indicate that cytokines might be a common mediator of tissue regression during molting induced by diverse methods, although the pattern of induction is different. Thus, a high dose of dietary zinc seems to induce reproductive regression via the upregulation of cytokines and chemokines, the suppression of feed intake, and the increase in serum corticosterone, resulting finally in the apoptosis of reproductive tissues.