Injection Of Pituitary Extract Research Articles

Study on Induced Breeding in Oreochromis niloticus (Nile Tilapia) through Hypophysation Technique

Producing fish to spawn by artificial methods is called induced breeding. Oreochromis niloticus (Nile tilapia) are induced to spawn by the injection of pituitary extract, a process called Hypophysation. Hypophysation is a technique of induced breeding in Oreochromis niloticus by injecting pituitary gland extract. In hypophysation, the pituitary gland extract is injected into intramuscular/pectoral fin region of fish. The gland induces the fish to breed. When the pituitary gland of the same species is used, the hypophysation is called homoplastic. Induced breeding is practised in a wide variety of species, with Hypophysation we can get pure seeds in large number in all seasons. With this technique we can produce hybrid varieties of species. This technique shows high fertilization rate (~85%) and Hatching rate (~80%) this study explodes about procedure and result of hypophysation technique.

Metabolite profiling of the post-ovulatory oocytes of the common carp, Cyprinus carpio: A 1H NMR-based metabolomics approach

A metabolomics study was conducted to investigate the molecular bases of oocyte over-ripening in common carp, Cyprinus carpio from a metabolic point of view. The ovulation was induced in fish brooders by intramuscular injection of pituitary extract and oocytes were collected four times post-ovulation with 30 min intervals. A set of 32 metabolites were identified on the NMR spectra of the oocytes, which mainly included energy-linked metabolites, amino acids, methylated metabolites and citric acid cycle (TCA) intermediates. PCA and PLS-DA models clearly separated the post ovulations times, indicating the effects of post-ovulation time on oocyte metabolome content. Based on the loading plot outputs, 15 metabolites including tryptophan, cysteine, AMP, tyrosine, valine, creatine phosphate (PCr), ATP, leucine, inosine, malate, acetate, TMAO, glucose, fumarate and lysine had more effects on the separation of post ovulation times. According to the results of metabolite profiling, the concentrations of glutamine, alanine, tryptophan, lysine and cysteine mostly significantly (P < 0.01) increased at 90 and 120 min post-ovulation. The concentrations of PCr, ATP, inosine and guanosine were relatively stable until 60 min post-ovulation, while significantly (P < 0.01) decreased at 90 and 120 min post ovulation. The TCA metabolites succinate, malate and fumarate significantly (P < 0.01) elevated at 90 and 120 min post-ovulation. AMP concentrations remained relatively unchanged until 30 min and then progressively decreased with time post ovulation (P < 0.01). The concentrations of lactate showed significant elevations at 90 and 120 min post ovulation (P < 0.01). In conclusion, the energetic potentials of the oocytes reduced with time post ovulation. There were apparent elevations in the concentrations of free amino acids, which may be associated with the onset of proteolytic activities in the post ovulatory oocytes. In addition, we found some changes in the apoptotic-related metabolites, which may support the results of previous studies regarding the oxidative stress and following apoptosis in post ovulatory oocytes of fish.

Effect of the Reduction in the Frequency of Injections of Pituitary Extracts During a Superovulation on the Embryo Collection Results in Cows in Algeria

The aim of this study is to verify the efficacy of a superovulation treatment with 3 injections of pituitary extracts (Stimufol®) 24 hours apart and compare the results to a standard protocol (8 injections 12 hours apart), with a total dose of 500 μg. Twenty-three embryos collections were conducted after two superovulation protocols, the first with 8 injections 12 hours apart and decreasing doses (n = 11, group 1) and the second 3 injections at constant doses 24 hours of interval (n = 12, group 2). The average number of lutea corpora obtained is 9.1 per cow for group 1 and 10.4 in group 2. A total number of 140 embryos were collected with an average of 6.08 per cow. Among the embryos collected, 46 and 56 per cent were transferable respectively in group 1 and 2. A little quality gain was recorded in group 2 with an average transferable embryo of 3.8 against 2.2 for group1. In our study we observed the same efficacy for both treatments with a little quality improvement (almost one transferable embryo) when reducing the number of injections at the time of superovulation treatment, the same gain minimal in elite cows can have a significant economic impact.

Successful, induced propagation of African catfish (Clarias gariepinus) by ovarian lavage with sperm and hormone mixture

The in vitro fertilisation method is widespread in fish aquaculture. In general, calculated doses of different hormones are injected intramuscularly or intraperitoneally, by using a syringe and needle, to induce ovulation and spermiation. We tested a novel approach to delivering sperm and hormone together to eggs, in African catfish, and concluded that sperm plus carp pituitary extract injection into the ovary lobes (through the oviduct) leads to successful ovulation and recovery of developing embryos in all of the experimental conditions applied. These results indicate that injected spermatozoa with suspended carp pituitary are stored in the ovary lobes for up to 10h without loss of biological activity and that the ovulated eggs could be fertilised by the injected sperm, after being released from the body cavity. In addition, the seminal fluid was an appropriate delivery vehicle for the ovulation inducing hormone (i.e. carp pituitary) via ovarian lavage.

Changes in ovarian gene expression profiles and plasma hormone levels in maturing European eel (Anguilla anguilla); Biomarkers for broodstock selection

Complete sexual maturation of European eels (Anguilla anguilla) in captivity can only be achieved via injections with gonadotropins. For female eels this procedure takes 4–6months and the response ranges from “unresponsive” to final maturation and ovulation. Reproductive success could be significantly increased via early selection of responders based on predictive markers and minimally invasive sampling methods. To get a better understanding of the genetic background of ovarian maturation of the European eel we performed a pilot deep-sequencing transcriptome analysis of ovarian tissue derived from a yellow eel, a prepubertal silver eel and a post-spawning matured eel. Two key players in steroidogenesis were strongly correlated with advanced sexual maturation, namely P450c17 and liver receptor homolog-1, suggesting that blood plasma steroids might qualify as minimally invasive markers for early detection of responders. Since the predictive value of plasma sex steroid levels for final maturation of the European eel had not yet been carefully examined, we performed an extensive artificial maturation trial. Farmed silver eels were treated with pituitary extracts and sampled at multiple time intervals. Expression of steroidogenesis-related genes in ovarian tissue of responding and non-responding eels after four weekly injections with pituitary extract was compared using a custom-built microarray and RNAseq. Increased expression of 17β-hsd1 was strongly linked to sexual maturation. Blood plasma levels of sex steroids were measured using ELISAs. We show that a 2.5-fold increase in blood-plasma estradiol level after 4 weekly pituitary extract injections is a strong predictor of final sexual maturation of female European eel.

Swimming suppresses hepatic vitellogenesis in European female silver eels as shown by expression of the estrogen receptor 1, vitellogenin1 and vitellogenin2 in the liver

BackgroundWhen European silver eels (Anguilla anguilla) venture into the Atlantic Ocean for their 6,000 km semelparous spawning run to the Sargasso Sea, they are still in a prepubertal stage. Further sexual development appears to be blocked by dopaminergic inhibition of hypothalamus and pituitary activity. Recently, we found that swimming for several weeks in freshwater stimulated the incorporation of fat droplets in the oocytes. So, it was hypothesized that long term swimming in seawater would release the inhibition further and would also stimulate the production of vitellogenin by the liver.MethodsFor this study a swim-flume was constructed to allow simulated migration of migratory female silver eels for 3 months (1,420 km) in natural seawater at 20 degrees C. Primers were designed for polymerase chain reactions to measure the mRNA expression of estrogen receptor 1 (esr1), vitellogenin1 (vtg1) and vitellogenin2 (vtg2) genes in the liver of European female silver eels.ResultsIn comparison to resting eels, swimming eels showed a diminished expression of esr1, vtg1 and vtg2 in the liver. They also had lower plasma calcium (Ca; indicative of vitellogenin) levels in their blood. This showed that vitellogenesis is more strongly suppressed in swimming than in resting eels. However, when eels were subsequently stimulated by 3 weekly carp pituitary extract injections, the expression of the same genes and plasma levels of Ca strongly increased in both groups to similar levels, thus equalizing the initial differences between resting and swimming.ConclusionsIt is concluded that vitellogenesis remains suppressed during resting and even more during swimming. The fact that swimming stimulates fat deposition in the oocytes but suppresses vitellogenesis indicates that these events are separated in nature and occur sequentially. Swimming-suppressed vitellogenesis may imply that in nature eels undergo vitellogenesis and final maturation near or at the spawning grounds.

Effect of Postmanufacturing Processing and Shipping of Luteinizing Hormone Releasing Hormone Analog on Induced Ovulation for Production of Channel Catfish Female × Blue Catfish Male Hybrid Fry

AbstractThe potential effects of postmanufacturing processing and shipping of luteinizing hormone releasing hormone analog (LHRH‐a) on induced ovulation for the production of hybrids of female channel catfish Ictalurus punctatus × male blue catfish I. furcatus were evaluated. There were no differences in efficacy regarding postmanufacturing bottling. Hand‐weighed LHRH‐a gave results similar to those of LHRH‐a reconstituted in water, aliquoted, and relyophilized. Shipping without dry ice reduced the number of fry per kilogram of female body weight by 51–87% . Theoretically, dry ice is not necessary when shipping powdered LHRH‐a, but these results indicate that to ensure efficacy (as indicated by total fry production) shipping LHRH‐a on dry ice would be a best management practice. Hybrid embryos were successfully produced, and LHRH‐a injections are a viable alternative to carp pituitary extract injections for producing hybrid catfish embryos.

Partial development of the steroidogenic ultrastructural features in degenerative corpora lutea after a single injection of pituitary extract in the Western painted turtle ( Chrysemys picta)

Pituitary glands were removed from sexually mature female turtles ( Chrysemys picta) and they were injected intraperitoneally (i.p.) into other mature females of the same species (experimental). In addition mature females of the same species received saline injection only (controls). Initially all the turtles used in this study were steroidogenically inactive with corpora lutea already undergoing luteolysis (degeneration) as these turtles had ovioposited their eggs approximately 2 weeks earlier. Forty-eight hour post injection the corpora lutea were removed from the control and experimental turtles. In the experimental turtles, the lutein granulosa cells developed ultrastructural features such as tubular and cisternal smooth endoplasmic reticulum (SER) and mitochondria with tubular cristae associated with lipid droplets. However, the controls maintained degenerative corpora lutea without steroidogenic ultrastructural features. The circulating progesterone (Pro) levels in the experimental turtles were significantly higher than the controls ( P < 0.049). Although the 48 h development of steroidogenic ultrastructural features in the lutein granulosa cells was only partial in development, the effect of the pituitary taken from the inactive donor triggered an activating process within a short period, clear evidence of gonadotropic effect on the inactive corpora lutea. The present data offer interesting information on the short-term effect of gonadotropins during the non-reproductive period. This information may have useful implication under natural conditions particularly during the onset of a new reproductive cycle where the ovary is still inactive.

Growth hormone: Historical notes

A brief review of important contributions to our present knowledge of growth hormone is given. In 1887 it had been noted that a pituitary tumor was present in most patients with acromegaly. Even at the beginning of the 20. Century relationship between growth disorders and the pituitary was contested. From 1908 pituitary surgery became established treatment in GH hypersecretion. In 1922 it was demonstrated that injection of pituitary extract to animals caused excessive growth and soon after the opposite: removal of the pituitary caused growth retardation. A huge number of studies on the effects of GH were subsequently reported as were trials with GH treatment. They were impeded by failure to recognize the impact of species specificity of GH. After this issue was clarified in 1957, treatment with human growth hormone proved effective. In 1985 it was realized that Creutzfeldt-Jakob's disease might be transmitted through human growth hormone. At this time recombinant GH had become available. In 1971 the structure of human GH was established. In the same period both GH releasing and inhibiting hormones were identified and an analogue of somatostatin had evolved into the first effective pharmacological treatment for acromegaly.

An in vitro system for the long-term tissue culture of juvenile rainbow trout (Oncorhynchus mykiss) testis

In vitro cultivation of fish gonad fragments continues to be an important experimental approach to answer both fundamental and applied scientific questions. The aim of the present investigation was to test whether juvenile rainbow trout (Oncorhynchus mykiss) testes cultured for a week or more were physiologically competent. Trout testis fragments (approximately 1 mm2) were placed on pieces of flat, culture plate insert filter, in a drop of liquid medium (modified Leibovitz's L-15), and floated on the medium surface in a multi-well culture plate. Culture plates were covered and incubated in air at 15 degrees C. Three different endpoints were used to test whether cultured testis fragments remained healthy and functional. First, a comparison of the histological appearance of testis fragments cultured in vitro for different periods up to 8 days with intact testes indicated that no differences were evident. Secondly, testis fragments incubated in medium supplemented with bovine calf serum (BCS) at concentrations of 2.5% and 25% BCS had significantly greater proliferation of interstitial and spermatocyst cells as measured by nuclear 5-bromo-2'-deoxyuridine incorporation. Finally, testis fragments cultured for 5 days and transplanted back into the donor fish underwent precocious sexual maturity in response to a regime of salmon pituitary extract injections and produced fertile sperm as determined by their ability to successfully fertilize eggs. These experiments demonstrate the utility of this method for in vitro culture of juvenile rainbow trout testis fragments.

The GH-IGF-I axis and diabetes complications.

Over 60 yr ago, Houssay demonstrated that removal of the pituitary improved the diabetes of animals and made them particularly sensitive to insulin (Carbohydrate metabolism. BA Houssay. N Engl J Med 1936; 214: 971–975), and Young reported that permanent diabetes could be produced by injection of pituitary extract (Permanent diabetes produced by pituitary (anterior lobe) injections. FG Young. Lancet 1937; ii: 372–374). Rolf Luft and colleagues in Sweden translated the Houssay model to humans by performing hypophysectomy in the hope of ameliorating vascular complications (Hypophysectomy in man. R Luft, H Olivecrona, B Sjogren. Nord Med 1952; 47: 351–354). At the same time, remission of diabetic retinopathy was reported in a woman with long-standing type 1 diabetes when she developed postpartum anterior pituitary insufficiency (Sheehan's syndrome) (The Houssay phenomenon in man: recovery from retinopathy in a case of diabetes with Simmonds' disease. JE Poulsen. Diabetes 1953; 2 : 7–12). In the 1960s and 1970s, there was considerable enthusiasm for pituitary ablation in the treatment of proliferative retinopathy, an approach happily supplanted by retinal photocoagulation.

Evaluation of sturgeon follicle-stimulating hormone, luteinizing hormone, estradiol, progesterone and testosterone in Acipenser persicus serum to identify fertile broodstock

Summary Various methods have been proposed for the selection of suitable sturgeon broodstock for artificial reproduction. In this study, serum levels of reproductive hormones, namely sturgeon homologues of follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), estradiol (E2) and progestrone (P), were measured by radioimmunoassay (RIA) in 36 mature broodstock before and after injection of pituitary extract, and the results correlated with sex, age and oocyte polarization index (p). Mature males were significantly younger than females, and showed significant differences in serum P and T levels following pituitary extract injection, although not before that. A significant difference was found between LH, E2 and T before and after injection in the female group. There was also a significant difference between serum levels of T and P in the fertile and infertile groups. Stepwise discriminant function analysis was used to discriminate between fertile and infertile groups. The function had 93,8% overall correct classification, i.e. amongst 100 female broodstock fish, almost 94% of them would be correctly allocated to the relevant group. So the proposed model provides a reliable method for selecting suitable broodstock fish.

Seasonal variations in sperm production and sperm quality in male winter flounder, Pleuronectes americanus : the effects of hypophysectomy, pituitary replacement therapy, and GnRH-A treatment

The pattern of sperm release and changes in sperm quality were investigated throughout the normal spawning season of male winter flounder, Pleuronectes americanus (Walbaum), caught in Conception Bay, Newfoundland, Canada in April and November 1994. In addition, the impact of hormonal manipulation on sperm production and sperm quality was examined through gonadotropin-releasing hormone analogue (GnRH-A) treatment, hypophysectomy, and pituitary extract injection – pituitary replacement therapy (PRT). While a lengthy period of spermiation, lasting ∼6 months (December to July), can be detected in some males, the major period for sperm release occurred in May to July in the summer spawning season. The spawning period started with peak levels of plasma testosterone (T) and 11 ketotestosterone (11-KT). By late July sperm production fell rapidly along with a notable deterioration in sperm motility. While GnRH-A treatment advanced the period of sperm release, and the milt became diluted and increased in volume, the treatment did not increase total sperm output over the spawning season. In hypophysectomized male flounder, by contrast, milt was more concentrated and sperm production declined along with reduced plasma androgen levels during the spawning season. Increased plasma T and 11-KT levels in hypophysectomized males followed PRT in December and January. Moreover, PRT appeared to be responsible for advancement of spermiation in a large number of prespawning males, which released an increased amount of milt. Finally, these studies indicated that both sperm motility and egg fertilization rates remained unchanged after these hormonal treatments.

Think globally, act locally: the making of a mouse mammary gland.

On a phylogenetic scale of organ development the mammary gland is a recent acquisition. It was introduced 200 million years ago with the appearance of mammals to provide nourishment to the newborn in the form of milk. The mammary gland is characterized by a unique dependence on hormonal signals for terminal differentiation, which is attained only after pregnancy. At the time of birth, the anlage consists of a few rudimentary ducts in the vicinity of the nipple. Pronounced ductal outgrowth and branching commences at puberty, and in pregnancy an expanded lobulo-alveolar compartment develops. Functional differentiation of the secretory epithelium coincides with parturition and large amounts of milk are produced and secreted during lactation. After weaning of the young, the entire alveolar epithelial compartment is remodeled to resemble a virgin-like state. With each pregnancy, a new round of lobulo-alveolar development occurs. During the past 100 years, intensive efforts have been made to understand the endocrine control of mammopoiesis and lactogenesis. Classical research on endocrine ablated animals firmly established that ovarian steroids and pituitary peptide hormones are mandatory and sufficient for breast development and lactation. In 1900, Halban first established that mammary growth is controlled by the ovary (Halban 1900). He demonstrated that ovariectomy caused mammary regression, and that transplanted ovaries prevented the castration atrophy of mammary glands. Twenty-eight years later, Stricker and Grueter induced mammary development and milk secretion artificially in castrated virgin rabbits by injection of pituitary extract (Stricker and Grueter 1928). In 1933, Riddle, Bates, and Dykshorn purified the respective pituitary hormone (Riddle et al. 1933) and named it prolactin(PRL). In the last several years, the ability to delete genes from the mouse genome has allowed us to identify genetic components of mammary gland development. Molecular insight into the underlying genetic framework and signaling networks of the developing tissue has been gained through experimental manipulations of tissues from wild-type and knockout mice. Two distinct, yet braided, developmental concepts have unfolded. First, discrete signaling networks activated by systemic endocrine hormones induce mammopoiesis. Secondly, some of these signals are relayed through reciprocal interactions between the epithelium and the stroma. Table 1 contains those genes whose elimination from the mouse genome results in impaired mammary gland development. Among these genes are some of the ‘‘usual suspects’’ but also some previously unidentified players. Each mutation affects specific and distinct aspects of mammary development. These knockout mice not only confirmed the involvement of hormonal signaling but also provided tools to identify the tissue compartment that receives and executes these signals.

Chapter 9 Isolation of Fish Sperm Flagella

Publisher Summary This chapter describes methods for purifying axonemes and the focus is on preparing intact flagella to analyze their membrane proteins and reactivate them to study axonemal motility mechanisms. Fish spermatozoa offer a good source from which to isolate and purify large amounts of vertebrate flagella and axonemes. Sperm from two teleost species, trout and carp, have been used extensively to study the mechanisms involved in the initiation of sperm motility. During the spawning season, an adult male trout produces a large number of sperm. In carp, sperm production can be induced throughout the year by repetitive injection of pituitary extracts. Trout and carp spermatozoa are quite simple and their flagella contain a classical 9+2 axoneme without periaxonemal structures. The absence of an acrosomal vesicle decreases the risk of contamination of the preparation with proteases. Trout sperm motility can be inhibited by media containing large amounts of potassium or having a low Ph. Carp sperm activation is controlled by a decrease in the external osmotic pressure and is not dependent on the external ions. It is advisable to include different protease inhibitors in extraction buffer (IMEN), as proteasomes are present on trout sperm plasma membrane. For trout, the water should be aerated with bubbling and frequently renewed. Chlorine must be avoided. For injection or sperm collection, fish must be anesthetized. For carp, spermiation can be induced throughout the year by intraabdominal injection of carp pituitary extract. Only a fraction of the sperm obtained from a single trout can be used directly for axoneme isolation. Therefore, for biochemical studies, some of the sperm may be frozen in liquid nitrogen for later use. The chapter discusses isolation of axonemes and flagella, and thereafter the demembranation and ATP reactivation of isolated carp sperm flagella.

Fertility of triploid backcross progeny, (GengoroubunaCarassius auratus cuvieri ♀ × carpCyprinus carpio ♂)F11 ♀ × carp or gengoroubuna ♂

The fertility of triploid backcross progeny, (gengoroubunaCarassius auratus cuvieri ♀ × carpCyprinus carpio ♂) F1 ♀ × gengoroubuna or carp ♂ is reported. In females, spawning could be induced by the injection of pituitary extracts. The fertilization rates were from 52.4 to 88.6%, and free swimming rates from 2.6 to 34.4%. Many normal larvae were obtained after crossing with the sperm of gengoroubuna. However, after crossing with sperm of the loachMisgurnus anguillicaudatus, most of the embryos died before hatching and no normal larvae were obtained.

Elevation of sex steroids and inhibition of PDP-glucuronyltransferase are out of phase during gonadal maturation in the common carp

1. Plasma sex steroid concentrations, onset of gonadal maturation, and hepatic microsomal UDP-glucuronyltransferase (UDPGT) activities were followed under natural temperature and photoperiod in outdoor tanks, and under controlled laboratory temperature and photoperiod regimens in common carp ( Cyprinus carpio). 2. Decreased activity of UDPGT was out of phase with elevations in plasma testosterone and 17β-estradiol during gonadal maturation. 3. Injection of pituitary extract induced final gonadal maturation and transient elevations (within 24 hr) of both plasma sex steroid concentrations and UDPGT activities. 4. There were no simple relationships between plasma sex steroid concentrations and activity of hepatic microsornal UDPGT in common carp.

Hormonal changes during induced ovulation of the carp, Cyprinus carpio

The changes in plasma concentrations of eight steroids (testosterone, testosterone glucuronide, estradiol, estradiol glucuronide, 17-hydroxyprogesterone, 17,20β-dihydroxy-4-pregnen-3-one (17,20βP), deoxycorticosterone, and cortisol) have been followed in three individual carp, Cyprinus carpio, during ovulation induced by carp pituitary extract. Deoxycorticosterone and estradiol glucuronide were not detectable and small amounts of 17,20βP were found only in one fish. A priming injection of pituitary extract, administered 24 hr before ovulation, stimulated an increase in testosterone 3–6 hr later, followed by an increase in estradiol. The second injection of pituitary extract given 12 hr after the priming dose, resulted in a second peak of testosterone which was accompanied by a peak of testosterone glucuronide, a conjugate which was usually formed in only small amounts following the priming dose. A sharp peak of 17-hydroxyprogesterone followed the second stimulation and it is suggested that this, rather than 17,20βP, may be the natural inducer of maturation in carp. Although cortisol levels showed large variations, they appeared to be stimulated by the priming dose of pituitary extract. Levels of all steroids, except for cortisol, fell rapidly after ovulation.

Observation of Early Development of Fertilized Ova Superovulated by Pituitary Extract Injection in Rat.

Observation of Early Development of Fertilized Ova Superovulated by Pituitary Extract Injection in Rat.

Induced breeding of the sabalo, Prochilodus platensis Holmberg

Maturation and ovulation were induced in female Prochilodus platensis with multiple homologous pituitary extract injections (total dose 0.9 to 1.4 hypophysis/kg) or single injections of pituitary extract combined with human chorionic gonadotrophin ((1 hypophysis + 500 I.U.)/kg). One of the females given the latter treatment produced viable eggs. Latency time from injection to ovulation was 9 h (at 28°C). Hatching occurred 12 h after fertilization. Survival after 30 days of rearing was 96%.