Reliability of intraperitoneal injections in fish

Résumé L'hormone thyréotrope¹²⁵I fut injectée en doses variées dans le péritoine de Cyprins. Les pertes de substance par écoulement dépassèrent le 50% des doses. Les meilleures rétentions furent obtenues avec de petits volumes, injectés à travers l'intestin, après bouchage du cloaque avec du liquide silicone.     

Transgenesis in fish

Gene transfer into fish embryo is being performed in several species (trout, salmon, carps, tilapia, medaka, goldfish, zebrafish, loach, catfish, etc.). In most cases, pronuclei are not visible and microinjection must be done into the cytoplasm of early embryos. Several million copies of the gene are generally injected. In medaka, transgenesis was attempted by injection of the foreign gene into the nucleus of oocyte. Several reports indicate that the injected DNA was rapidly replicated in the early phase of embryo development, regardless of the origin and the sequence of the foreign DNA. The survival of the injected embryos was reasonably good and a large number reached maturity. The proportion of transgenic animals ranged from 1 to 50% or more, according to species and to experimentators. The reasons for this discrepancy have not been elucidated. In all species, the transgenic animals were mosaic. The copy number of the foreign DNA was different in the various tissues of an animal and a proportion lower than 50% of F1 offsprings received the gene from their parents. This suggests that the foreign DNA was integrated into the fish genome at the two cells stage or later. An examination of the integrated DNA in different cell types of an animal revealed that integration occurred mainly during early development. The transgene was found essentially unrearranged in the fish genome of the founders and offsprings. The transgenes were therefore stably transmitted to progeny in a Mendelian fashion. Southern blot analysis revealed the presence of possible junction fragments and also of minor bands which may result from a rearrangement of the injected DNA. In all species, the integrated DNA appeared mainly as random end-to-end concatemers. In adult trout blood cells, a small proportion of the foreign DNA was maintained in the form of non-integrated concatemers, as judged by the existence of end fragments. The transgenes were generally only poorly expressed. The majority of the injected gene constructs contained essentially mammalian or higher vertebrates sequences. The comparison of the expression efficiency of these constructs in transfected fish and mammalian cells indicates that some of the mammalian DNA sequences are most efficiently understood by the fish cell machinery. Chloramphenicol acetyl transferase gene under the control of promoters from Rous sarcoma virus, and human cytomegalovirus, was expressed in several tissues of transgenic fish. Chicken -crystallin gene was expressed in several tissues of transgenic fish. Rainbow trout growth hormone cDNA driven by the Rous sarcoma virus promoter was expressed in transgenic carps leading to a faster growth of these animals. The antifreeze protein gene from flounder was expressed in transgenic salmon. These data indicate that transgenesis in fish is relatively easy but that fish gene sequences must be preferably used to obtain a good expression of the transgenes. Fish is a good biological model, specially for developmental studies and it is an increasing part of human food. For these reasons, transgenesis in fish is most likely to be more and more practised in the coming years.     

Transgenesis in fish

Gene transfer into fish embryo is being performed in several species (trout, salmon, carps, tilapia, medaka, goldfish, zebrafish, loach, catfish, etc.). In most cases, pronuclei are not visible and microinjection must be done into the cytoplasm of early embryos. Several million copies of the gene are generally injected. In medaka, transgenesis was attempted by injection of the foreign gene into the nucleus of oocyte. Several reports indicate that the injected DNA was rapidly replicated in the early phase of embryo development, regardless of the origin and the sequence of the foreign DNA. The survival of the injected embryos was reasonably good and a large number reached maturity. The proportion of transgenic animals ranged from 1 to 50% or more, according to species and to experimentators. The reasons for this discrepancy have not been elucidated. In all species, the transgenic animals were mosaic. The copy number of the foreign DNA was different in the various tissues of an animal and a proportion lower than 50% of F1 offsprings received the gene from their parents. This suggests that the foreign DNA was integrated into the fish genome at the two cells stage or later. An examination of the integrated DNA in different cell types of an animal revealed that integration occurred mainly during early development. The transgene was found essentially unrearranged in the fish genome of the founders and offsprings. The transgenes were therefore stably transmitted to progeny in a Mendelian fashion. Southern blot analysis revealed the presence of possible junction fragments and also of minor bands which may result from a rearrangement of the injected DNA. In all species, the integrated DNA appeared mainly as random end-to-end concatemers. In adult trout blood cells, a small proportion of the foreign DNA was maintained in the form of non-integrated concatemers, as judged by the existence of end fragments. The transgenes were generally only poorly expressed. The majority of the injected gene constructs contained essentially mammalian or higher vertebrates sequences. The comparison of the expression efficiency of these constructs in transfected fish and mammalian cells indicates that some of the mammalian DNA sequences are most efficiently understood by the fish cell machinery. Chloramphenicol acetyl transferase gene under the control of promoters from Rous sarcoma virus, and human cytomegalovirus, was expressed in several tissues of transgenic fish. Chicken delta-crystallin gene was expressed in several tissues of transgenic fish.(ABSTRACT TRUNCATED AT 400 WORDS)     

Bacterial RNA synthesis in frog auricles after intraperitoneal injection of bacteria

Résumé Des organes de plantes ou d'animaux mis dans une suspension bactérienne synthétisent du RNA bactérien. Ce phénomène que nous avons appelé transcession est dû au transfert de DNA spontanément cédé par les bactéries vivantes aux cellules des organismes supérieurs. Dans le présent travail, il est démontré que le même phénomène peut avoir lieu naturellement lorsqu'une grenouille est sujette à une infection bactérienne.

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We thank MissA. Cattaneo, MissJ. Henry for excellent technical assistance. This work was supported by a grant from the Fonds National Suisse de la Recherche Scientifique.     

Electro-actography in fresh-water fish

Zusammenfassung Es wird eine Methode beschrieben, das Aktogramm von Fischen und anderen Süsswasserbewohnern zu bestimmen aufgrund der vom Autor entdeckten, von diesen Organismen erzeugten äusseren schwachelektrischen Felder.     

Daily melatonin injections inhibit short-day-induced testicular regression in hamsters

Summary Daily injections of melatonin were found to retard testicular regression in hamsters exposed to LD 10:14, if the injections occurred in the morning (i.e., 0.5 h after lights on), but not if they occurred in the afternoon (i.e., 6 h after lights on). These results indicate that appropriately timed injections of melatonin can at least partially block the inhibitory effects of short days on gonadal activity in the photoperiodic hamster.We wish to thank Susan Losee and Susan Stice for excellent technical assistance and Gary Ellis for his comments. This investigation was supported by NIH grants HD-09885, HD 12622 and Research Career Development Award HD-00249 (F. W. T.) from the National Institute of Child Health and Human Development.     

Feeding elicited in sheep by intrahypothalamic injections of PGE1

Résumé Une augmentation de la prise de nourriture a lieu après injection intrahypothalamique du PGE₁ en dose de 21 nmoles, chez les brebis. La température de la cavité abdominale n'est pas affectée par ces injections, sauf une légère augmentation au moment de l'injection et qui peut provenir du maniement des animaux.

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We are grateful to Dr.John E. Pike for the gift of PGE₁.

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This research was supported in part by a Grant-in-aid of the National Science Foundation, Grant No. GB-28836.     

Effects of hypothalamic injections of 5,6-dihydroxytryptamine on thermoregulation in rats

Summary 5,6-Dihydroxytryptamine, a serotonin depletor, infused directly into the anterior hypothalamus of rat's brain, produced an increase in both heat production and heat loss (as indicated by changes in peripheral circulation) at temperatures of 8, 15 and 22°C. The rectal temperature of these treated rats remained constant.This work was supported by grants from National Science Council of Taiwan and J. Aron Charitable Foundation (USA). The author is grateful to Dr C. Y. Chai and Mr C. C. Wei for their support.     

Comparison of the effectiveness of intramuscular and intraperitoneal ACTH in the rat

Summary Injection of 1–24 ACTH is more effective by the i.m. than i.p. route. Large doses are required to induce consistent maximal adrenal corticosterone secretion.Supported by grants from the National Institute of Arthritis, Metabolism and Digestive Diseases, National Institutes of Health, Bethesda, Maryland, USA.     

Urinary output of rats in response to subcutaneous injections of adrenaline

Zusammenfassung Es werden der Effekt einer s.c. Injektion von Adrenalin auf die renale Ausscheidung von Wasser und Elektrolyten bei Ratten sowie Versuche über die Korrelation verschiedener Dosis abhängiger, renaler Adrenalin-Wirkungen beschrieben.     

Experimental hypertension elicited by injections of methyl cellulose

Résumé Des injections sous-cutanés d'une solution de méthyl-cellulose provoquent dans la rate une vraie hypertension artérielle maligne. Les animaux utilisés eurent fréquemment des dèmes et des ascites et toujours des lésions artérielles dans le cur, les reins et ailleurs.

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This study was supported by grants H-2703 and H-4327 from the United States Public Health Service.