In vitro biosynthesis of juvenile hormone in larval honey bees: Comparison of six media

Rachinsky, Anna; Hartfelder, Klaus

doi:10.1007/s11626-996-0014-x

Cited by 26 publications

(17 citation statements)

References 18 publications

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“…Ovaries were dissected in honey bee saline, and the ovarioles were individualized in honey bee culture medium (Rachinsky and Hartfelder, 1998) before tracheae and the peritoneal sheath were manually removed as far as possible with the aid of number 5 watchmaker forceps (Dumont). The individualized ovarioles were fixed for 2 h in a glutaraldehyde/paraformaldehyde (2%/2%) mix in cacodylate buffer (50 mM, pH 7.4).…”

Section: General Histologymentioning

confidence: 99%

“…• C in 5-bromo-2'deoxy-uridine (BrdU) labeling reagent (Roche) at a final concentration of 50 mM in honey bee tissue culture medium (Rachinsky and Hartfelder, 1998) supplemented with 0.1% DMSO. After two brief rinses in fresh medium, the ovarioles were individualized and cleared of trachea and peritonial sheath before fixation (overnight at 4…”

Section: Brdu Immunocytochemistrymentioning

confidence: 99%

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Ovariole structure and oogenesis in queens and workers of the stingless beeMelipona quadrifasciata(Hymenoptera: Apidae, Meliponini) kept under different social conditions

Tanaka¹,

Santana²,

Hartfelder³

2009

Apidologie

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-The high variability in the reproductive biology of stingless bees makes them very amenable for comparative studies with other eusocial bee taxa. We investigated the structural organization of the ovaries of Melipona quadrifasciata queens and workers kept under different social conditions by analyzing their general histology, mitotic activity, and microfilament organization. The overall dynamics of ovarian activity were similar in the two castes, and at emergence their ovarioles contained a previtellogenic follicle. Stingless bees and honey bees differ in the structural organization in the lower germarium, but they have in common synchronized mitotic activity and putative germ line stem cells in the terminal filament. Unlike honey bees, stingless bee workers lay trophic eggs in addition to reproductive eggs. The overall similarities in oogenesis between the two taxa suggest that the decision to form trophic eggs should only occur in the late stages of oogenesis.worker reproduction / follicle development / actin cytoskeleton / ovary histology

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Section: General Histologymentioning

confidence: 99%

Section: Brdu Immunocytochemistrymentioning

confidence: 99%

Ovariole structure and oogenesis in queens and workers of the stingless beeMelipona quadrifasciata(Hymenoptera: Apidae, Meliponini) kept under different social conditions

Tanaka¹,

Santana²,

Hartfelder³

2009

Apidologie

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“…Transfer these to a dish containing honey bee culture medium (recipe and preparation steps as described in Rachinsky and Hartfelder, 1998) or a commercial insect culture medium (e.g.…”

Section: Dissection and Fixation Of Ovariolesmentioning

confidence: 99%

Standard methods for physiology and biochemistry research inApis mellifera

Hartfelder

Bitondi

Brent

et al. 2013

Journal of Apicultural Research

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SummaryDespite their tremendous economic importance, and apart from certain topics in the field of neurophysiology such as vision, olfaction, learning and memory, honey bees are not a typical model system for studying general questions of insect physiology. The reason is their social lifestyle, which sets them apart from a "typical insect" and, during social evolution, has resulted in the restructuring of certain physiological pathways and biochemical characteristics in this insect. Not surprisingly, the questions that have attracted most attention by researchers working on honey bee physiology and biochemistry in general are core topics specifically related to social organization, such as caste development, reproductive division of labour and polyethism within the worker caste. With certain proteins playing key roles in these processes, such as the major royal jelly proteins (MRJPs), including royalactin and hexamerins in caste development, and vitellogenin in reproductive division of labour and age polyethism, a major section herein will present and discuss basic laboratory protocols for protein analyses established and standardized to address such questions in bees. A second major topic concerns endocrine mechanisms underlying processes of queen and worker development, as well as reproduction and polyethism, especially the roles of juvenile hormone and ecdysteroids. Sensitive techniques for the quantification of juvenile hormone levels circulating in haemolymph, as well as its synthesis by the corpora allata are described. Although these require certain instrumentation and a considerable degree of sophistication in the analysis procedures, we considered that presenting these techniques would be of interest to laboratories planning to specialize in such analyses. Since biogenic amines are both neurotransmitters and regulators of endocrine glands, we also present a standard method for the detection and analysis of certain biogenic amines of interest. Further questions that cross borders between individual and social physiology are related to energy metabolism and thermoregulation. Thus a further three sections are dedicated to protocols on carbohydrate quantification in body fluid, body temperature measurement and respirometry. Métodos estándar para la investigación de la fisiología y bioquímica de Apis mellifera ResumenA pesar de su enorme importancia económica, y aparte de ciertos temas en el campo de la neurofisiología, tales como la visión, el olfato, el aprendizaje y la memoria, las abejas no son un sistema modelo típico para el estudio de cuestiones generales sobre la fisiología de los insectos. La razón de ello es su forma de vida social, lo que las diferencia de un "insecto típico" y que durante la evolución social, se ha traducido en la reestructuración de ciertas vías fisiológicas y bioquímicas propias de este insecto. Como era de esperar, las preguntas que han atraído mayor atención por parte de los investigadores que trabajan en la fisiología y la bioquímica de la abeja melífera, son en general tema...

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“…For probing the ecdysone response in ovaries in vitro, batches of 50 L5F3 worker larvae per sample were dissected and their ovaries were incubated for 6 h at 34.5°C in 250 µL larval honey bee medium (Rachinsky and Hartfelder, 1998) supplemented with makisterone A (final conc. 5 × 10 -7 M) (Hartfelder et al, 1995).…”

Section: Northern Blot Analysis Of Honey Bee Sdr Expressionmentioning

confidence: 99%

A member of the short-chain dehydrogenase/reductase (SDR) superfamily is a target of the ecdysone response in honey bee (Apis mellifera) caste development

2004

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-Many aspects in caste polyphenism result from hormonally controlled differential gene expression. A DDRT-PCR screen for ecdysteroid-regulated genes in ovaries revealed a set of ESTs coding for metabolic enzymes. For a cDNA encoding a short-chain dehydrogenase/reductase (SDR) we obtained the complete coding sequence (246 amino acids), revealing the protein motifs typical of insect SDRs. Its initially high expression in early fifth-instar larvae vanished in prepupae. Expression levels in worker larvae were higher than in queen larvae, suggesting negative regulation by the caste-specific ecdysteroid titer. This finding was confirmed by in vitro exposure of competent worker ovaries to makisterone A. In contrast to whole body RNA extracts, two SDR transcripts were detected in the ovaries. Both had their expression downregulated by makisterone A. Hormonal regulation and tissue-specific expression pattern makes this SDR an interesting enzyme for comparative molecular studies on social insect caste polyphenisms.Apis mellifera / differential display PCR / ecdysteroid / alcohol dehydrogenase / caste polyphenism

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In vitro biosynthesis of juvenile hormone in larval honey bees: Comparison of six media

Cited by 26 publications

References 18 publications

Ovariole structure and oogenesis in queens and workers of the stingless beeMelipona quadrifasciata(Hymenoptera: Apidae, Meliponini) kept under different social conditions

Ovariole structure and oogenesis in queens and workers of the stingless beeMelipona quadrifasciata(Hymenoptera: Apidae, Meliponini) kept under different social conditions

Standard methods for physiology and biochemistry research inApis mellifera

A member of the short-chain dehydrogenase/reductase (SDR) superfamily is a target of the ecdysone response in honey bee (Apis mellifera) caste development

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