A gonadotropin releasing hormone analog induces spermiation in intact and hypophysectomized frogs, Rana esculenta

The sperm-releasing activity of a gonadotropin releasing hormone (GnRH) agonist, Buserelin (GnRH) and hypophysis homogenate (PD) preparations was studied in intact and hypophysectomized (PDX) frogs, Rana esculenta. In addition, human chorion gonadotropin (hCG) was tested in PDX animals, and GnRH antagonist (GnRHA) treatments were carried out in intact and PDX animals, in combination with the hormonal injections. GnRH or PD treatments were able to elicit spermiation in intact and PDX animals. While GnRH, injected 24 h later, was again effective in inducing spermiation in intact animals, this was not the case in PDX frogs. GnRHA counteracted GnRH effects in intact frogs. Moreover, in PDX animals GnRHA injections counteracted the sperm-releasing activity induced by hCG or GnRH, but failed to inhibit sperm-releasing activity induced by PD homogenate.     

Embryonic chick muscle produces an FGF-like activity

Normal and pathological formation of blood vessels is of considerable interest both in terms of basic scientific processes and clinical applications. Angiogenic events in the adult are likely to represent persistence of developmental mechanisms, and embryos are therefore a suitable experimental model for these processes. Among embryonic tissues, muscle is particularly appropriate for investigation, since it is highly vascularised from early stages. There are a number of competing explanations of how this process is controlled. Bioassays offer advantages over conventional molecular localisation techniques, in that they reveal the presence of active processed forms of the molecules under study, rather than non-processed forms, or non-translated meassages. Using these techniques, we report here that embryonic chick muscle, taken from the stages at which blood vessels are forming, produces an angiogenic activity on the chick chorioallantoic membrane (CAM), and transforms NR6 cells in soft agar. Basic fibroblast growth factor (bFGF) is shown to be angiogenic on the CAM in the same way, and also transforms NR6 cells (NR6 cells lack functional epidermal growth factor/transforming growth factor-a receptors, and are believed to respond only to bFGF in this way). Anti-bFGF removes the transforming activity of the embryonic muscle. We conclude that this represents evidence that embryonic chick muscle is producing an FGF-like molecule which is capable of acting as an angiogenic agent at the appropriate times in development.     

Secretogranin III: a diabetic retinopathy-selective angiogenic factor

Secretogranin III (Scg3) is a member of the granin protein family that regulates the biogenesis of secretory granules. Scg3 was recently discovered as an angiogenic factor, expanding its functional role to extrinsic regulation. Unlike many other known angiogenic factors, the pro-angiogenic actions of Scg3 are restricted to pathological conditions. Among thousands of quantified endothelial ligands, Scg3 has the highest binding activity ratio to diabetic vs. healthy mouse retinas and lowest background binding to normal vessels. In contrast, vascular endothelial growth factor binds to and stimulates angiogenesis of both diabetic and control vasculature. Consistent with its role in pathological angiogenesis, Scg3-neutralizing antibodies alleviate retinal vascular leakage in mouse models of diabetic retinopathy and retinal neovascularization in oxygen-induced retinopathy mice. This review summarizes our current knowledge of Scg3 as a regulatory protein of secretory granules, highlights its new role as a highly disease-selective angiogenic factor, and envisions Scg3 inhibitors as “selective angiogenesis blockers” for targeted therapy.     

Ovarian LDH activity in gonadotropin-treated immature rats.

Lactate dehydrogenase (LDH) activity was studied in the ovaries of immature rats treated with pregnant mare serum gonadotropin (PMS). LDH activity increased sharply at 36 h after PMS injection in the ovarian tissue as well as in the blood. It was suggested that the increase of LDH activity in the ovary may be related to its increasing ability to secrete estrogen.     

Hypothalamic tyrosine hydroxylase activity,plasma gonadotropin and prolactin levels after aminooxyacetic acid in ovariectomized rats

Summary Plasma concentrations of gonadotropin, prolactin and hypothalamic tyrosine hydroxylase (TH) activity were measured in ovariectomized rats treated with aminooxyacetic acid (AOAA), a drug which elevates brain GABA levels. Hypothalamic TH activity was significantly increased with a significant decrease in prolactin (Prl) release. Plasma levels of gonadotropins were not modified by AOAA. These results support an inhibitory action of GABA on Prl release possibly mediated through hypothalamic dopamine.Supported by grants from Indian Council of Medical Research (ICMR), New Delhi. RIA kits for the estimation of LH, FSH and Prl were kindly supplied by Dr A.F. Parlow, NIAMDD-NIH, Bethesda, Maryland, USA. GNB is a UGC research fellow.     

Ovarian LDH activity in gonadotropin-treated immature rats

Summary Lactate dehydrogenase (LDH) activity was studied in the ovaries of immature rats treated with pregnant mare serum gonadotropin (PMS). LDH activity increased sharply at 36 h after PMS injection in the ovarian tissue as well as in the blood. It was suggested that the increase of LDH activity in the ovary may be related to its increasing ability to secret estrogen.     

Hypothalamic tyrosine hydroxylase activity, plasma gonadotropin and prolactin levels after aminooxyacetic acid in ovariectomized rats

Plasma concentrations of gonadotropin, prolactin and hypothalamic tyrosine hydroxylase (TH) activity were measured in ovariectomized rats treated with aminooxyacetic acid (AOAA), a drug which elevates brain GABA levels. Hypothalamic TH activity was significantly increased with a significant decrease in prolactin (Prl) release. Plasma levels of gonadotropins were not modified by AOAA. These results support an inhibitory action of GABA on Prl release possibly mediated through hypothalamic dopamine.     

Immunological demonstration of large quantities of glycogen or of a glycogen-like substance in human embryonic colon cells and in colon carcinomas by means of rabbit antisera raised against a strain of Escherichia coli 013]

Rabbit antisera raised against a strain of E. coli 013, with a strong antiglycogen activity, were tested on human fetal and normal adult colons, on colon carcinomas, and on colon tumor cells in culture (HT29). Only very rare granules were present in adult normal colons when tested with the immunofluorescence method. In faetal colons, in 12 out of 14 carcinomas, and on HT29 cells, the immunofluorescent reactions were similar to those observed in normal liver. The reactions were negative after previous treatment with alpha-amylase. They were inhibited with glycogen, with phenol-alcohol, perchloric, and trichloroacetic extracts from faetal colons, and with a tumor trichloroacetic extract. The extracts precipitated with anti-E. coli 013 antisera. They had a strong inhibiting activity in a radioimmunoassay test with labeled glycogen. The extracts from normal adult colons did not precipitate with the antisera and they had no inhibiting activity in either immunofluorescence and radioimmunoassay tests.     

Different salt concentration requirement of nuclear and cytoplasmic Mn++-stimulated poly(A) synthetase activities from rat liver

Summary Mn⁺⁺-stimulated poly(A) synthetase activity of dialyzed ammonium sulfate fraction (0–50% saturation) of nuclear extracts from rat liver is dependent on salt addition to the incubation mixture. The corresponding soluble cytoplasmic activity is inhibited by the salt concentration required for nuclear activity.     

A molecular recognition hypothesis for nonpeptides: Na+ K+ ATPase and endogenous digitalis-like peptides

The molecular recognition hypothesis for peptides is that binding sites of ligands and their receptors are encoded by short, complementary segments of DNA. A corollary hypothesis for nonpeptide ligands posited here is that peptide replicas may be encoded by the DNA segment complementary to the receptor binding sites for nonpeptides. This corollary was tested for digitalis, a family of cardiotonic and natriuretic steroids including ouabain. A hexapeptide (ouabain-like peptide, OLP) complementary to a ouabain binding site on sodium/potassium dependent adenosine triphosphatase (Na⁺ K⁺ ATPase) exhibited activity in a digitalis bioassay. Antisera to the complementary peptide (OLP) stained the neurohypophysis in an immunocytochemical procedure. The complementary peptide was found to share an identical 4-amino acid region with the 39-amino acid glycopeptide moiety of the vasopressin-neurophysin precursor. This glycopeptide was isolated from pituitary extracts; it exhibited digitalis-like activity in the submicromolar range and cross-reacted with complementary peptide antibodies. Another digitalis-like substance with high activity also was detected in the extracts. These results demonstrate that the vasopressin-neurophysin glycopeptide has digitalis-like activity. Moreover, the findings are consistent with the hypothesis that peptide mimetics of nonpeptides are encoded in the genome. Received 23 November 1998; received after revision 18 January 1999; accepted 19 February 1999     

Appearance of growth-inhibiting activity (G1 chalone) during ontogenetic development of rat and chick epidermis in vivo and in vitro.

A comparative study was carried out between the stage of embryonic development of epidermis and its content of growth-inhibitory activity (G1 chalone). Injections of aqueous extracts from keratinized fetal rat or chick embryo epidermis led to a depression of DNA-synthesis in adult mouse epidermis, whereas extracts from undifferentiated epidermis did not contain such an activity.     

Cell cycle patterns of thymidylate synthetase and 5,10-methylenetetrahydrofolate polyglutamates in cultured mouse hepatoma cells

Summary The regulation of thymidylate synthetase activity was investigated throughout the first cell cycle after release from an isoleucine block in synchronous cultures of mouse hepatoma (Hepa) cells. Activity in cell extracts increased with the onset of S phase and the increased activity was attributed to a parallel increase in enzyme concentration as determined by titration with tritiated fluorodeoxyuridylate. The polyglutamate chain length of reduced folate cofactors, which could also influence thymidylate synthetase activity, was unchanged.Acknowledgments. Supported by grant CA 22754, awarded by the National Cancer Institute, DHEW.     

Appearance of growth-inhibiting activity (G1 chalone) during ontogenetic development of rat and chick epidermis in vivo and in vitro

Summary A comparative study was carried out between the stage of embryonic development of epidermis and its content of growth-inhibitory activity (G₁ chalone). Injections of aqueous extracts from keratinized fetal rat or chick embryo epidermis led to a depression of DNA-synthesis in adult mouse epidermis, whereas extracts from undifferentiated epidermis did not contain such an activity.This investigation was supported by the Deutsche Forschungsgemeinschaft.     

Ornithine decarboxylase, S-adenosyl-L-methionine decarboxylase and arginine decarboxylase from Mycobacterium bovis (BCG)

Ornithine decarboxylase (ODC), S-adenosyl-L-methionine decarboxylase (AMDC) and arginine decarboxylase (ADC) activities were detected for the first time in extracts of Mycobacterium bovis (BCG). All the decarboxylases differed from corresponding known bacterial decarboxylases in that: a) ODC did not require GTP for activity; b) ODC was not inhibited by any known inhibitor of bacterial ODCs; c) AMDC and ADC did not require Mg2+-ion for activity and were not markedly inhibited by any known inhibitor of the decarboxylases of other bacteria.     

Neurovascular development and links to disease

The developing central nervous system (CNS) is vascularized via ingression of blood vessels from the outside as the neural tissue expands. This angiogenic process occurs without perturbing CNS architecture due to exquisite cross-talk between the neural compartment and invading blood vessels. Subsequently, this intimate relationship also promotes the formation of the neurovascular unit that underlies the blood–brain barrier and regulates blood flow to match brain activity. This review provides a historical perspective on research into CNS blood vessel growth and patterning, discusses current models used to study CNS angiogenesis, and provides an overview of the cellular and molecular mechanisms that promote blood vessel growth and maturation. Finally, we highlight the significance of these mechanisms for two different types of neurovascular CNS disease.     

Increased angiogenesis in diabetes.

Rats with streptozotocin-induced diabetes mellitus showed a 3.4-4.5 times increased angiogenic response following mast-cell activation in situ as compared with age-matched normal controls. The test tissue used was the mesenteric window, which we have previously exploited as a quantitative angiogenesis assay. In the present study two independent techniques for quantifying the angiogenic response showed essentially the same result. The finding of a pathologically increased angiogenic reaction in the diabetic animals is noteworthy since some of the most harmful complications of diabetes in man relate to proliferative vascular lesions.     

CNS stimulation effect on the sexual maturation of the female rat

Summary In the immature rat, CNS stimulants administration to pregnant mare serum gonadotropin (PMSG) primed rats resulted in significant (p<0.01) ovarian and uterine hypertrophy when compared to animals treated with PMSG only. Meanwhile precocious puberty was produced by pentylenetetrazol treatment alone. The results of this experiment may indicate that administration of CNS stimulants has a specific action on the release of endogenous gonadotropin.Supported by NIH grant no. RR 8111.National Institute of Health, MBS Graduate Trainee.     

Ornithine decarboxylase,S-adenosyl-L-methionine decarboxylase and arginine decarboxylase fromMycobacterium bovis (BCG)

Summary Ornithine decarboxylase (ODC), S-adenosyl-L-methionine decarboxylase (AMDC) and arginine decarboxylase (ADC) activities were detected for the first time in extracts ofMycobacterium bovis (BCG). All the decarboxylases differed from corresponding known bacterial decarboxylases in that: a) ODC did not require GTP for activity; b) ODC was not inhibited by any known inhibitor of bacterial ODCs; c) AMDC and ADC did not require Mg²⁺-ion for activity and were not markedly inhibited by any known inhibitor of the decarboxylases of other bacteria.     

Increased angiogenesis in diabetes

Summary Rats with streptozotocin-induced diabetes mellitus showed a 3.4–4.5 times increased angiogenic response following mast-cell activation in situ as compared with age-matched normal controls. The test tissue used was the mesenteric window, which we have previously exploited as a quantitative angiogenesis assay. In the present study two idependent techniques for quantifying the angiogenic response showed essentially the same result. The finding of a pathologically increased angiogenic reaction in the diabetic animals is noteworthy since some of the most harmful complications of diabetes in man relate to proliferative vascular lesions.     

Search for the neurodepressing hormone in a stomatopod,Squilla mantis

Summary Crude extracts ofSquilla mantis eyestalks have no neurodepressing activity in theProcambarus bouvieri bioassay. However, partially purified extracts show a very high level of neurodepressing hormone with the same characteristics as the decapod hormone.This investigation was supported by grant No. 192 from the Consejo Nacional de Ciencia y Tecnologia, and by a grant from Fondo de Fomento Educativo. We are very thankful to Dr Victor Anaya for performing the bioassays.