Aldehyde Mixture Research Articles

Inhibition of glycolytic enzymes by endogenous aldehydes: a possible relation to diabetic neuropathies

Endogenous saturated and unsaturated aldehydes were found in significant elevations in serum of diabetic humans and rats. These compounds, originating from the lipid peroxidation processes, are shown here to be potent inhibitors of the glycolytic enzymes, phosphofructokinase and glyceraldehyde-3-phosphate dehydrogenase. The inhibition process is non-competitive and progressive. The aldehyde mixture, when supplemented to the standard rat diet at 1 100 ratio, caused nerve damage that is reminiscent of diabetic polyneuropathies.

Cellular relations in mouse circumvallate taste buds.

The fine structure of the taste buds of circumvallate papillae of two strains of mice was studied by electron microscopy. Mice anesthetized with ketamine were perfused through the heart with a double aldehyde mixture in cacodylate buffer and the tissues embedded in Epon. Semi-serial sections were employed. The morphology and relationships of cell types are consistent with the majority of descriptions of mammalian taste buds served by the ninth cranial nerve. Cells of type II are particularly well documented, as the stages in their origin, maturation and degeneration could be followed. Significant differences, however, relate to cell type I. These cells contain large dense-cored granules, contrasted with the more irregular and somewhat larger dark granules of the type I cells in the rabbit. These granules do not produce a dense homogenous product for the pore, as seen in the rabbit. Rather the pore substance consists of small, empty vesicles in a diffuse dark matrix. These granules are only moderately larger than the dense-cored vesicles of the type III cells. All features of the type III cell were demonstrated, although no instance of a complete cell was seen in any section. No significant differences were noted between the two strains of mice. Intimate proximity of a nerve to a cell nucleolus, suggestive of a trophic pathway, is illustrated.

Tetracycline administration increases protein (presumably procollagen) synthesis and secretion in periodontal ligament fibroblasts of streptozotocin-induced diabetic rats.

Streptozotocin-induced, insulin-deficient diabetic adult rats were daily administrated either minocycline or a chemically-modified non-antimicrobial tetracycline (CMT) by oral gavage for a 3-week time period; untreated diabetic and non-diabetic rats served as controls. On day 21, all rats received an intravenous injection of 3H-proline followed by perfusion fixation with an aldehyde mixture at 20 minutes and 4 hours after isotope injection. The upper and lower mandibles of these rats were dissected and processed for quantitative electron microscopic autoradiography to study 3H-proline utilization by fibroblasts in the periodontal ligament (PDL) of molars. In the non-diabetic controls, at 20 min after 3H-proline injection, radioprecursor was incorporated by the Golgi-RER system of PDL fibroblasts. At the 4-h time period, most of the label was present over the collagen fibers around these cells. In contrast, PDL fibroblasts in the untreated diabetic rats showed marked abnormalities ultrastructurally and minimal uptake (20 min) and secretion (4 h) of labeled proline. At both time periods, in both minocycline- and CMT-treated diabetic rats, fibroblasts were structurally more normal and the radioprecursor was localized in the fibroblasts and the PDL matrix in a pattern similar to that seen in the control rats. These results suggest that the diabetes-induced structural abnormalities and suppression of synthesis and secretion of protein (presumably collagen and its precursor) by PDL fibroblasts can be restored to near-normal by administration of a tetracycline and that this effect is mediated by a non-antimicrobial property of this family of antibiotics.

Ultrastructural immunogold localization of prostaglandin endoperoxide synthase (cyclooxygenase) to non-membrane-bound cytoplasmic lipid bodies in human lung mast cells, alveolar macrophages, type II pneumocytes, and neutrophils.

Lipid bodies are non-membrane-bound, lipid-rich cytoplasmic inclusions that occur in many mammalian cell types. Because lipid bodies are more prominent in cells associated with inflammation and are repositories of arachidonyl-phospholipids, a role for lipid bodies in the oxidative metabolism of arachidonic acid to form eicosanoids has been suggested. To evaluate further whether lipid bodies, in addition to serving as non-membranous sources of substrate arachidonate, are involved in eicosanoid formation, we used cells isolated from human lung to investigate the intracellular localization of prostaglandin endoperoxide (PGH) synthase (cyclooxygenase), the key initial, rate-limiting enzyme in the formation of prostaglandins and thromboxanes. Isolated lung cells containing a mixture of mast cells, alveolar macrophages, Type II alveolar pneumocytes, and neutrophils from short-term cultures were fixed in suspension in a dilute aldehyde mixture, post-fixed in osmium tetroxide, stained en bloc with uranyl acetate, dehydrated in a graded series of alcohols, and embedded in Epon. A post-embedding immunogold procedure was used with a primary PGH synthase monoclonal antibody and 20-nm gold-conjugated secondary antibody to demonstrate enzyme locations. Specificity controls were also done. We found PGH synthase in lipid bodies of human lung mast cells, alveolar macrophages, Type II alveolar pneumocytes, and neutrophils. Specific secretory and lysosomal granules and plasma membranes did not express PGH synthase. Specificity controls, including omission of the primary antibody or substitution with an irrelevant antibody, were negative. Absorption of the specific PGH synthase antibody with purified solid-phase PGH synthase resulted in a marked reduction of label in lipid bodies of all four cell types. These findings establish the presence of PGH synthase in lipid bodies of human lung mast cells, alveolar macrophages, Type II alveolar pneumocytes, and neutrophils and, in concert with previous studies, suggest that these cytoplasmic lipid-rich organelles may be non-membrane sites of eicosanoid formation.

Tetracycline administration increases collagen synthesis in osteoblasts of streptozotocin-induced diabetic rats: a quantitative autoradiographic study.

Streptozotocin-induced, insulin-deficient diabetic rats were administrated either minocycline (MC) or a chemically modified non-antimicrobial tetracycline (CMT) by oral gavage for a 3-week period; untreated diabetic and nondiabetic rats served as controls. On day 21, all rats received an intravenous injection of 3H-proline followed by perfusion fixation with an aldehyde mixture at 20 minutes and 4 hours after isotope injection. The parietal bones of these rats were dissected and processed for quantitative electron microscopic autoradiography to study 3H-proline utilization by osteoblasts. At 20 minutes after 3H-proline injection, radioprecursor was incorporated by the Golgi-RER system of the osteoblasts in the periosteal surface of the control rats. At the 4-hour time period, most of the label was present over the collagen fibers of the osteoid. In contrast, the flattened bone-lining cells in the untreated diabetic rats showed minimal uptake (20 minutes) and secretion (4 hours) of labeled proline. In both MC and CMT-treated diabetic rats, the radioprecursor was localized in the osteoblasts and osteoid matrix in a pattern similar to that seen in the control rats at both 20 minutes and 4 hours after isotope injection. Labeling of the osteoid by the radioprecursor was greater as a result of CMT treatment than during minocycline treatment. These results suggest that the diabetes-induced suppression of synthesis and secretion of protein (presumably collagen and its precursor) by osteoblasts can be restored to near-normal levels by administration of tetracycline(s) and that this effect is mediated by a non-antimicrobial property of these antibiotics.

生理的組織接着剤（ベリプラストＰ）の骨欠損填塞後の骨形成に関する組織学的解析

This paper describes the effects of fibrin adhesive material (Beriplast P ®: BP) on bone induction in bone defects formed by surgical operation. Using adult Sprague-Dawley strain rats, bone defects were formed in the cortical bones of femurs with a dental round bar 1 mm in diameter, and immediately filled with BP. Bone defects without BP served as controls.On days 1, 4, 7, 14, 30, 60, and 120 after operation, rats were fixed by transcardiac perfusion with an aldehyde mixture, and dissected bones were examined by light, transmission, and scanning-electron microscopy.In the control bones without BP, the defective regions werefirst filled with blood clots (day 1) and then with granulated tissues containing numerous fibroblasts and macrophages (day 4). In BP-filled defects, the presence of BP and surrounding fibrous tissues significantly inhibited the formation of blood clots and granulated tissues, so bone formation from the surrounding cortical bone margins was induced faster than in control bones. In addition, granulated tissues were rapidly replaced by normal bone marrow in the BP-filled defects. Because BP is biological adhesive, at 4 days after operation, most of the BP was already absorbed, but no dead space was left in the defected regions. At 2 weeks after operation, the defects were completely filled with newly-formed bones, in which an active bone remodelling by osteoblasts and osteoclasts occurred.These results show that BP is an effective biological tool in filling bone defects and in inducing new bone formation.

Tetracycline administration restores osteoblast structure and function during experimental diabetes

Osteopenia is a recognized complication of diabetes mellitus in humans and experimental animals. We recently found that tetracyclines prevent osteopenia in the streptozotocin-induced diabetic rat and that this effect was associated with a restoration of defective osteoblast morphology (Golub et al., 1990). The present study extends these initial ultrastructural observations by assessing osteoblast function in the untreated and tetracycline-treated diabetic rats. After a 3-week protocol, non-diabetic control and diabetic rats, including those orally administered a tetracycline, minocycline (MC), or a non-antimicrobial tetracycline analog (CMT), were perfusion-fixed with an aldehyde mixture; the humeri were dissected and processed for ultracytochemical localization of alkaline phosphatase (ALPase) and Ca-ATPase activities. Some rats from each experimental group received an intravenous injection of 3H-proline as a radioprecursor of procollagen, and the humeri were processed for light microscopic autoradiography. In addition, the osteoid volume in each experimental group was quantitatively examined by morphometric analysis of electron micrographs. During the diabetic state, active cuboidal osteoblasts in the endosteum of control rats were replaced by flattened bone-lining cells that contained few cytoplasmic organelles for protein synthesis (Golgi-RER system), and active transport (mitochondria). Treating diabetic rats with MC, and even more so with CMT, appeared to "restore" osteoblast structure. During diabetes, bone-lining cells incorporated little 3H-proline or secreted little labeled protein and produced only a very thin osteoid layer. Tetracycline administration to the diabetics increased both the incorporation of 3H-proline by osteoblasts and their secretion of labeled protein toward the osteoid matrix, in a pattern similar to that seen in the non-diabetic controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Paraffin Sections of Nervous Tissue Supra Vitally Stained with Methylene Blue: A New, Reliable and Simple Fixation Technique

Two steps are recommended for fixation of central nervous tissue after supravital staining with methylene blue: application of ammonium heptamolybdate followed by a paraformaldehyde-glutaraldehyde mixture. The two fixatives complement each other so that both dye and tissue are optimally fixed. Molybdate renders the dye water and alcohol insoluble, the aldehyde mixture conditions the tissue for embedding in paraffin. This technique is a suitable alternative to conventional whole mount and frozen sectioning methods.

A new sealant for knitted Dacron prostheses: Minimally cross-linked gelatin

There has been a resurgence of interest in the concept of presealing high-porosity knitted Dacron prostheses with an absorbable biologic material. Such a material should provide reliable porosity control, preferably reducing water porosity from 2000 ml/cm2/min to less than 50 ml/cm2/min. It should not interfere with the fibrous and vascular ingrowth that securely anchors the developing pseudointima. In previous studies, we have examined fibrin glue and two forms of aldehyde cross-linked insoluble collagen used as Dacron sealants. We concluded that delayed resorption of the sealant as seen with glutaraldehyde cross-linked insoluble collagen results in undesirable healing characteristics, particularly lack of adhesion between pseudointima and the luminal surface of a prosthesis. This study examines a new sealant. Soluble collagen (gelatin) is treated to reduce the number of free amino groups available for aldehyde cross-linking. It is then weakly cross-linked with an aldehyde mixture and applied to a knitted Dacron prosthesis. Water porosity studies have confirmed satisfactory porosity control. Both rat subcutaneous and canine circulatory implants for 6 months reveal relatively rapid and complete sealant resorption without undesirable modification of the normal healing process of knitted Dacron.

A new sealant for knitted dacron prostheses: Minimally cross-linked gelatin

There has been a resurgence of interest in the concept of presealing high-porosity knitted Dacron prostheses with an absorbable biologic material. Such a material should provide reliable porosity control, preferably reducing water porosity from 2000 ml/cm2/min to less than 50 ml/cm2/min. It should not interfere with the fibrous and vascular ingrowth that securely anchors the developing pseudointima. In previous studies, we have examined fibrin glue and two forms of aldehyde cross-linked insoluble collagen used as Dacron sealants. We concluded that delayed resorption of the sealant as seen with glutaraldehyde cross-linked insoluble collagen results in undesirable healing characteristics, particularly lack of adhesion between pseudointima and the luminal surface of a prosthesis. This study examines a new sealant. Soluble collagen (gelatin) is treated to reduce the number of free amino groups available for aldehyde cross-linking. It is then weakly cross-linked with an aldehyde mixture and applied to a knitted Dacron prosthesis. Water porosity studies have confirmed satisfactory porosity control. Both rat subcutaneous and canine circulatory implants for 6 months reveal relatively rapid and complete sealant resorption without undesirable modification of the normal healing process of knitted Dacron.

The effects of chemical fixation on the permeability of frog mesenteric capillaries.

1. We have investigated the effects on microvascular permeability of two chemical fixatives, glutaraldehyde/formaldehyde mixture and osmium tetroxide, both of which are used widely in electron microscopy. 2. The permeability of single perfused frog mesenteric capillaries was assessed using the technique of Michel (1980) to estimate the hydraulic conductance (Lp) of the vessel walls and the effective osmotic pressure (sigma delta pi) which could be exerted across them by the neutral macromolecule, Ficoll 70. 3. In each experiment, Lp and sigma delta pi were estimated in the same capillary before and after chemical fixation with either an aldehyde mixture (n = 6) or 1% osmium tetroxide (n = 6). In the absence of serum proteins in the perfusate, the values of Lp recorded prior to fixation were high. The effect of both fixatives was to reduce Lp. With aldehydes the mean value (+/- S.E.M.) of Lp fell from 22.1 (+/- 6.3) x 10(-3) to 5.7 (+/- 2.0) x 10(-3) microns s-1 cmH2O-1. With osmium tetroxide the mean Lp fell from 29.1 (+/- 8.5) x 10(-3) to 8.6 (+/- 3.6) x 10(-3) microns s-1 cmH2O-1. 4. The perfusates contained the neutral macromolecule, Ficoll 70, at a concentration of 60 mg ml-1 which exerted an osmotic pressure of 35 cmH2O in a membrane osmometer. In the absence of plasma proteins in the perfusate, sigma delta pi was only a fraction of this perfusate oncotic pressure prior to fixation. After aldehyde treatment there was a small but insignificant decrease in sigma delta pi. Osmium tetroxide, however, increased sigma delta pi from a mean (+/- S.E.M.) of 6.3 (+/- 1.6) cmH2O before fixation to 16.4 (+/- 2.8) cmH2O after fixation, P less than 0.01 using a paired t test. 5. In three experiments, the reflection coefficient of the capillary wall to NaCl sigma NaCl, was measured using the technique of Curry, Mason & Michel (1976) before and after osmium fixation. Fixation raised sigma NaCl from a mean value (+/- S.E.M.) of 0.009 (+/- 0.001) to one of 0.017 (+/- 0.004). Although these changes may be interpreted as an increase in the contribution to the total Lp of the capillary wall of channels which are available only to water, the changes are too small for such a mechanism to account for the increase in reflection coefficient to Ficoll 70. 6. The reduction in Lp following chemical fixation could represent an increase in the hydraulic resistance of either the capillary wall itself, or in the tissues surrounding the vessel.(ABSTRACT TRUNCATED AT 400 WORDS)

Methyl enol ethers as artefacts in capillary gas chromatographic profiles of aldehyde dimethyl acetals

Methyl enol ethers are detected within the capillary gas chromatographic profiles of aldehyde dimethyl acetals derived from human red cell plasmalogens or from a synthetic aldehyde mixture. They arise from the dimethyl acetals via methanol elimination as artefacts of the injection technique. In a nutritional experiment, the probands showed a rising proportion of dimethyl acetals in the phospholipid fraction of their erythrocyte ghosts; this fact is seen as an effect of the lipid composition of the milk fat administered.

A preparation method for observing intracellular structures by scanning electron microscopy.

In order to observe intracellular structures by scanning electron microscopy, excess cytoplasmic matrix must be removed from the fractured surface of cells. Previously we reported an Osmium-DMSO-Osmium method devised for this purpose. This method is very effective in revealing intracellular structures, but requires osmium tetroxide for initial fixation with some consequent disadvantages. In the present study, a revised Osmium-DMSO-Osmium method is reported, in which an aldehyde mixture is used as the initial fixative instead of osmium tetroxide. As fixation is carried out by perfusion in this revised method, better preservation of fine structures is achieved than by the original method, especially in the central nervous tissue which tends to suffer from post-mortem degeneration. Moreover this method can be applied to cytochemical studies of intracellular structures with a scanning electron microscope (SEM). In this study, acid phosphatase of lysosomes is demonstrated in a coloured SEM micrograph.

Attraction of Hylemya antiqua (Meigen) (Diptera: Anthomyiidae) in the field to host- produced oviposition stimulants and their nonhost analogues

VERNON, R. S., G. J. R. JUDD, J. H. BORDEN, H. D. PIERCE, JR., and A. C. OEHLSCHLAGER. 1981. Attraction of Hylemya antiqua (Meigen) (Diptera: Anthomyiidae) in the field to host-produced oviposition stimulants and their nonhost analogues. Can. J. Zool. 59: 872-881. Because of low, natural population levels, laboratory-reared Hylemya antiqua were employed in evaluating chemical attractants in the field. Of 13 chemicals (held in paraffin oil for slow release) tested in buried, partially enclosed water traps, dipropyl disulfide, propanethiol, dipropyl sulfide, propenyl propyl sulfide, 2-butyl propyl sulfide, dibutyl disulfide, and methyl butyl disulfide were attractive to H. antiqua. Males were approximately three times more responsive to these compounds than females. Females were twice as responsive to cut onions as males, and the rate of attraction increased for both sexes as the cut onions aged, and released more volatiles. Four combined onion-produced aldehydes (propionaldehyde, 2-methyl butanal, 2-methylpentanal, and trans-2-methyl-2-pentanal) were attractive to females but not to males. An aldehyde mixture plus a mixture of onion-produced thiopropyl-containing compounds was additive in attractiveness to males and females.

Aqueous aldehyde (Faglu) methods for the fluorescence histochemical localization of catecholamines and for ultrastructural studies of central nervous tissue.

Aqueous solutions combining a high concentration of formaldehyde (4%) with low concentrations of glutaraldehyde (0.5--01%) have been used to simultaneously localize amines by the formation of fluorescent products and to fix central nervous tissue for electron microscopy. The fluorescence reaction is produced by the aldehyde mixture at room temperature and the fluorescence is stable when the tissue is maintained in aqueous solution. This means that nerve cell bodies and terminal fields which contain catecholamines can be located accurately in vibratome sections at the light microscope level and, after further processing, can be examined under the electron microscope. With 1% glutaraldehyde in the aldehyde mixture, ultrastructural details are well preserved; there is no significant distortion of any component of the tissue. If vibratome or cryostat sections are dried against glass slides, the intensity of the fluorescence reaction is enhanced and the sections can be permanently mounted.

Distribution of Schwann cell cytoplasm and plasmalemmal vesicles (caveolae) in peripheral myelin sheaths. An electron microscopic study with thin sections and freeze-fracturing.

The mode of distribution of Schwann cell cytoplasm in the various regions of the peripheral myelin sheath is reviewed using freeze-fractured and thin sectioned nerves from bullfrogs, chickens and cats, perfused with an aldehyde mixture or fixedin situ with potassium permanganate.

Water-stable fluorophores, produced by reaction with aldehyde solutions, for the histochemical localization of catechol- and indolethylamines

The properties of a new fluorescence histochemical method for arylethylamines based on reaction with a mixture of 4% formaldehyde and 0.5% glutaraldehyde in aqueous solution are described. At room temperature the aldehyde mixture produced a well-localized fluorescence reaction in tissues, which, when examined microscopically in aqueous solution, was sufficiently intense for fine terminal noradrenergic axons to be seen. If the tissue was subsequently dried, the fluorescence intensity increased. At the same time as inducing the fluorophores, the aldehyde mixture fixed the tissue to a standard well suited for electron microscopy. It thus proved possible to locate amine containing cells in the fluorescence microscope and subsequently examine their ultrastructure. In aqueous models, the aldehyde mixture formed fluorescent products with adrenaline, noradrenaline, dopamine, dopa, 5-hydroxytryptamine and 5-hydroxytryptophan, but not with histamine or octopamine. The fluorescence induced in the aldehyde mixture remained stable if the tissue was subsequently transferred to saline or distilled water and when it was dehydrated in ethanol and cleared with xylene, benzene, chloroform or acetone.

Acid phosphatase and peroxidase in "resting" acinar cells of the major salivary glands of cats and their possible movement into secretory granules.

After fixation of perarterial perfusion using an aldehyde mixture, salivary tissues were prepared for ultrastructural cytochemistry of acid phosphatase or peroxidase. Great variations in the distributions of the reaction product occurred, often within the same cell. Acid phosphatase staining occurred not only in lysosomes and sometimes in a GERL system, but a diffuse cytoplasmic component was also found in submandibular central acinar cells and to a lesser extent in parotid acini and variable staining occurred in the secretory granules of these cells. Peroxidase was variably associated with rough endoplasmic reticulum in submandibular demilunar cells, parotid acini, and more strongly in some sublingual cells. The secretory granules of the latter were darkly stained, but in parotid granules there was variable staining and least staining occurred in the granules of submandibular demilunes. These results are thought to indicate that not all enzymes present in secretory granules have reached there by an elective secretory process. Sometimes they appear to have entered the granules haphazardly, possibly having been enzymes associated with intracellular cisternal channels for transport or metabolism of other secretory substances and ultimately to have passed into the cisternal channels by chance or as part of a natural removal of redundant material.

Dorsal column nuclei projections to the cerebellar cortex in cats as revealed by the use of the retrograde transport of horseradish peroxidase.

The existence of a cerebellar projection from the dorsal column nuclei (gracile and cuneate nuclei, DCN) has been proposed on electrophysiological grounds but questioned when studied with neuroanatomical techniques. The retrograde transport of horseradish peroxidase (HRP) has been used for the present study and provides anatomical evidence of a DCN-cerebellar pathway. In adult cats, 1 to 6 mul of 30% HRP were injected in pars intermedia of the anterior lobe (lobules IV-V), in paramedial lobule and in vermis of the anterior (lobules IV-V) and of the posterior lobe (lobule VII). After survival of 24 to 48 hours, all animals were perfused with a double aldehyde mixture and serial 40 mu sections through the medulla oblongata were incubated for visualization of HRP. In all cases, medullary nuclei known to project to the injected cortical regions of the cerebellum contained HRP-positive neurons mainly ipsilateral to the injection (e.g., external cuneate nucleus) or mainly contralateral to it (e.g., inferior olivary complex). Following ipsilateral injections in either the paramedian lobule or the pars intermedia, HRP-positive neurons in the cuneate nucleus were concentrated in its rostral portion where multipolar cells with radiating dendrites predominate. In contrast, none of the clusters region, in the caudal part of the cuneate nucleus, displayed HRP-positive granules. In cases in which the anterior vermis was injected a few labelled cells were present in the rostral part of the gracile nucleus but not in the clusters region of this nucleus. No labelling of DCN neurons was evident after posterior vermis injection. To compare the distribution of cells contributing to the DCN-cerebellar pathway with that of thalamic relay cells in the DCN, 0.5 to 3 mul of 30% HRP were injected in the nucleus ventralis posterolateralis of the thalamus in another series of cats. Contralateral to the thalamic injection, labelled cells were concentrated in the clusters region of the gracile and cuneate but rostrally in these nuclei they were scattered among unlabelled neurons. The preferential location in the DCN of cells which project to the cerebellum and of cells which project to the thalamus stresses the heterogeneous organization of these nuclei along the rostrocaudal axis. Further, the results indicate that regions of the DCN which have been distinguished on the basis of cytoarchitectonics (Kuypers and Tuerk, '64) and of afferents (Rustioni, '73, '74) differ also in their efferent projections.

Light and fine Structural Studies of Natural and Artificially Induced Egg Growth of Penaeid Shrimp1

ABSTRACTEgg differentiation in penaeids induced to undergo gonad maturation by eyestalk ablation has been examined with light and electron microscopes.Controls consisted of both wild brown and white shrimp (Penaeus aztecus and P. setiferus) caught in various stages of ovarian maturation. These animals were sacrificed and ovarian tissues were excised and fixed in a cold (4 C) aldehyde mixture, post‐fixed in osmic acid, dehydrated in acetone, and embedded in an epoxy resin. Experimental animals included laboratory‐reared, wild immature, and wild mature animals which had resorbed ovarian tissues. These animals were ablated with hot surgical clamps and sacrificed at various time intervals. Tissues were prepared as described for the controls.The development of ova from experimental animals was compared to control animals. In the stages early oocyte, cisternal phase, platelet phase, cortical rod phase, and mature oocyte, the experimental oocytes appeared the same as the control tissues. Though further studies (e.g., biochemical and zygote development) are needed, these preliminary fine structural studies are encouraging. While ablation, a mechanical process, may not be the technique of choice for maturing shrimp, it does indicate that manipulation of the endocrine system will be successful.