scholarly journals Migração espermática em suínos após inseminação artificial intrauterina profunda

2014 ◽  
Vol 66 (5) ◽  
pp. 1359-1366
Author(s):  
F.R.C.L. Almeida ◽  
V.A. Gheller ◽  
P.A. Auler ◽  
G.H.F.A. Moreira ◽  
R.B.C. Jardim ◽  
...  
Keyword(s):  
Phosphate Buffered Saline ◽  
Pbs Phosphate Buffered Saline

A inseminação artificial intrauterina profunda (IIP) é de grande importância para a indústria suinícola, em função do maior número de doses produzidas por reprodutores de alto mérito genético e da possibilidade da utilização de biotecnologias, como sêmen sexado e/ou congelado. Entretanto, necessita-se compreender com maior propriedade os mecanismos pelos quais os espermatozoides colonizam as tubas uterinas. Assim sendo, pretende-se com o presente experimento avaliar a existência ou não de migração intraperitoneal de espermatozoides inseminados profundamente em um dos cornos uterinos, mediante a obtenção de oócitos fertilizados no corno contralateral à inseminação e seccionado na base, na junção com o corpo do útero. Quatorze fêmeas pluríparas foram divididas em dois grupos experimentais, sendo que em um deles as fêmeas foram submetidas à secção da base de um dos cornos uterinos (Grupo Operado, n = 7), enquanto as do Grupo Controle (n = 7) não foram submetidas a nenhuma intervenção cirúrgica. Ambos os grupos foram submetidos à IIP, sendo as fêmeas abatidas 5±1,2 dias após a última inseminação. Os sistemas genitais das fêmeas foram coletados, dissecados e o número de corpos lúteos contados em ambos os ovários. A recuperação dos embriões foi feita por meio de lavagem das tubas e cornos uterinos com solução de PBS (Phosphate Buffered Saline), após o que se avaliou os fluidos coletados em lupa para a identificação de embriões. Em ambos os grupos experimentais, foram encontrados embriões nos segmentos do sistema genital de ambos os lados. Apenas uma fêmea apresentou embriões nos segmentos em somente um dos lados no grupo operado. Diante dos resultados aqui observados, concluiu-se que a migração espermática no suíno pode ocorrer tanto por via retrógrada pelo útero quanto por migração intraperitoneal. Estes achados certamente contribuirão para aumentar a eficiência da técnica de IIP, sendo de grande valia para o aprimoramento da indústria suinícola.

scholarly journals Tuned Cd2+ Selectivity: Showcase of Electronic and Regio-Effect of π-Extended Di-2-Picolylamine-Substituted Quinoline-Based Tolans

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 917
Author(s):  
Min-Sung Ko ◽  
P. Sankara Rao ◽  
Dong-Gyu Cho
Keyword(s):  
Metal Ions ◽  
Steric Hindrance ◽  
Limit Of Detection ◽  
Energy Difference ◽  
Turn On ◽  
Fluorescence Titration ◽  
Metal Selectivity ◽  
Substituted Effect ◽  
Phosphate Buffered Saline ◽  
Pbs Phosphate Buffered Saline

π-Extended di-2-picolylamine (DPA)-substituted 8-hydroxyquinoline (8-HQ) tolans (2) were synthesized for testing electronic and regio-effects. The electron-poor CN-tolan (2b) showed clear selectivity for Cd2+ (>>Zn2+) over other metal ions via turn-on fluorescence, while the electron-rich MeO-tolan (2a) displayed no clear metal selectivity. Furthermore, considering that there was no significant energy difference between the Cd2+ complexes of 1 and 2b, the intended regio-effect (7- vs. 5-substituted effect) did not induce steric hindrance. Thus, the regio-effect is mainly electronic. Considering the above, 2a and 2b constitute a complete showcase in which electronic and regio-effects modulate the metal selectivity. The fluorescence titration of 2b (10 mM) with Cd2+ showed that the limit of detection (LOD) of the Cd2+-selective 2b was 158 nM in PBS (phosphate-buffered saline) (10 mM, pH 7.2) containing 50% MeOH.

scholarly journals TÉCNICAS DE COLORAÇÃO CROMOSSÔMICA PARA ESTÁGIOS ESPECÍFICOS DA MATURAÇÃO NUCLEAR DE OÓCITOS BOVINOS

1994 ◽  
Vol 24 (3) ◽  
pp. 583-589 ◽  
Author(s):  
Alvaro Hernández Vignola ◽  
André do Prado ◽  
Alexandre Valente ◽  
Mara Batistella Rubin ◽  
Paulo Bayard Dias Gonçalves
Keyword(s):  
Phosphate Buffered Saline ◽  
Pbs Phosphate Buffered Saline

O presente trabalho foi conduzido com o objetivo de avaliar a eficácia de três técnicas de coloração nuclear de oócitos bovinos nos estágios de vesícula germinativa (VG), metáfase I (MI) e metáfase II (MII). Nestes três estágios de maturação nuclear, os oócitos, sem as células do cumulus, foram fixados em ácido acético:metanol (1:3) em lâminas (fixados em lâmina e corados com lacmóide; FLL) ou placa de petri (fixados em placa e corados com lacmóide; FPL) e corados com lacmóide a 1% em PBS (phosphate buffered saline). Os oócitos foram também distribuídos aleatoriamente a um terceiro grupo, no qual foram fixados em lâmina e corados com Giemsa (fixados em lâmina e corados com Giemsa; FLG). No estágio de VG, a percentagem de oócitos corretamente identificados no tratamento FLL (93,5%) foi significativamente maior do que nos tratamentos FPL (53,1%; χ²9,84; p-0,0017) e FLG (55,2%; χ2=9,03; p=0,0027). No entanto, a proporção de oócitos nos estágios de MI e MII precisamente corados com a técnica FLL (MI 41,7% e MII 41,9%) foram estatisticamente inferiores do que aquelas observadas nos tratamentos FPL (MI 90,0%, χ²13,25; p=0,0003, e MII 92.8%, χ2=12,45, p=0,0004) e FLG (MI 83,3%, χ2=10,69; p=0,0011, e MII 89,7%, χ²=12,24; p=0,0005). Assim sendo, os resultados demonstraram que a eficácia das técnicas de fixação e coloração dos oócitos bovinos depende do estágio de maturação nuclear que está em estudo.

scholarly journals Comparison of Vascular Perturbations in an Aβ-Injected Animal Model and in AD Brain

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Nattinee Jantaratnotai ◽  
Jae K. Ryu ◽  
Claudia Schwab ◽  
Patrick L. McGeer ◽  
James G. McLarnon
Keyword(s):  
Animal Model ◽  
Neuronal Damage ◽  
Molecular Layer ◽  
Prominent Feature ◽  
Model Of Alzheimer’S Disease ◽  
Intrahippocampal Injection ◽  
The Mean ◽  
Phosphate Buffered Saline ◽  
Pbs Phosphate Buffered Saline

The validity of amyloid-βpeptide (Aβ1–42) intrahippocampal injection, as an animal model of Alzheimer's disease (AD), has previously been considered in terms of inflammatory reactivity and neuronal damage. In this work, we have extended the testing of the animal model to vasculature by comparison of selected properties of microvesselsin vivowith those in human AD brain tissue. The injection of Aβ1–42, relative to control PBS (phosphate buffered saline), increased the mean number of microvessels and diminished the mean length of microvessels in the molecular layer of dentate gyrus. The animal model showed Aβ1–42, but not PBS, injection was associated with abnormalities in morphology of microvessels which were characterized as looping, fragmented, knob-like, uneven, and constricted. In particular, numbers of constricted microvessels, defined as vessels with diameters less than 3 μm, were considerably enhanced for Aβ1–42, compared to PBS, injection. In comparison, human AD brain demonstrated an elevated number of microvessels with a diminished mean length relative to nondemented (ND) brain. Additionally, microvessel perturbations in AD brain showed a similar pattern of morphological abnormalities to those observed in Aβ1–42-injected rat hippocampus. Constricted microvessels were a prominent feature of AD brain but were rarely observed in ND tissue. These results provide the first evidence that a peptide-injection animal model exhibits a commonality in perturbations of microvessels compared with those evident in AD brain.

Ultrastructure of developing visual cortical synapses in the cat superior colliculus

1991 ◽  
Vol 49 ◽  
pp. 156-157
Author(s):  
Caroline A. Miller ◽  
Laura L. Bruce
Keyword(s):  
Superior Colliculus ◽  
Phosphate Buffer ◽  
Receptive Fields ◽  
Visual Cortical Area ◽  
Cortical Synapses ◽  
Visual Cortical ◽  
And Function ◽  
Phosphate Buffered Saline ◽  
The Brain ◽  
Cat Superior Colliculus

The first visual cortical axons arrive in the cat superior colliculus by the time of birth. Adultlike receptive fields develop slowly over several weeks following birth. The developing cortical axons go through a sequence of changes before acquiring their adultlike morphology and function. To determine how these axons interact with neurons in the colliculus, cortico-collicular axons were labeled with biocytin (an anterograde neuronal tracer) and studied with electron microscopy.Deeply anesthetized animals received 200-500 nl injections of biocytin (Sigma; 5% in phosphate buffer) in the lateral suprasylvian visual cortical area. After a 24 hr survival time, the animals were deeply anesthetized and perfused with 0.9% phosphate buffered saline followed by fixation with a solution of 1.25% glutaraldehyde and 1.0% paraformaldehyde in 0.1M phosphate buffer. The brain was sectioned transversely on a vibratome at 50 μm. The tissue was processed immediately to visualize the biocytin.

Artifact-free electron microscopic immunocytochemistry on rat brain tissue

1991 ◽  
Vol 49 ◽  
pp. 272-273
Author(s):  
Veronika Burmeister ◽  
N. Ludvig ◽  
P.C. Jobe
Keyword(s):  
Microscopic Examination ◽  
Free Electron ◽  
Electron Microscopic Examination ◽  
Ultrastructural Localization ◽  
Rabbit Serum ◽  
Electron Microscopic ◽  
Electron Microscopic Immunocytochemistry ◽  
Phosphate Buffered Saline ◽  
Rat Brain Tissue ◽  
Immune Rabbit

Electron microscopic immunocytochemistry provides an important tool to determine the ultrastructural distribution of various molecules in both normal and pathologic tissues. However, the specific immunostaining may be obscured by artifactual immunoreaction product, misleading the investigator. Previous observations show that shortening the incubation period with the primary antibody from the generally used 12-24 hours to 1 hour substantially reduces the artifactual immunostaining. We now extend this finding by the demonstration of artifact-free ultrastructural localization of the Ca2/calmodulindependent cyclic nucleotide phosphodiesterase (CaM-dependent PDE) immunoreactivity in brain.Anesthetized rats were perfused transcardially with phosphate-buffered saline followed by a fixative containing paraformaldehyde (4%) and glutaraldehyde (0.25%) in PBS. The brains were removed, and 40μm sections were cut with a vibratome. The sections were processed for immunocytochemistry as described by Ludvig et al. Both non-immune rabbit serum and specific CaM-dependent PDE antibodies were used. In both experiments incubations were at one hour and overnight. The immunostained sections were processed for electron microscopic examination.

Modified gach technique vs critical point drying: Shrinkage analysis for SEM

1987 ◽  
Vol 45 ◽  
pp. 954-955
Author(s):  
B. Thompson ◽  
N. Sculov ◽  
R.E. Crang
Keyword(s):  
Critical Point ◽  
Calf Serum ◽  
Drying Shrinkage ◽  
Specimen Preparation ◽  
Cell Shrinkage ◽  
Whole Cells ◽  
Critical Point Drying ◽  
Prepared Material ◽  
Inverted Microscope ◽  
Phosphate Buffered Saline

The use of co-polymerized glutaraldehyde-carbohydrazide (GACH) was proposed for specimen preparation in scanning electron microscopy (SEM) as a means of avoiding dehydration in organic solvents, and to provide dimensionally stable biological specimens through a process of air-drying. It has been assumed that shrinkage of specimens prepared by the GACH technique should be less than that of conventionally-prepared material by critical point drying (CPD). In a previous study, Bell has reported significant shrinkage of whole cells for SEM. This report compares cell shrinkage in GACH and CPD preparations.Fibroblasts from newborn rats were grown on collagen-coated glass cover-slips (with alpha numeric grids etched onto the surface of the coverslips) in Eagle's minimum essential medium + 10% fetal calf serum for 7 d. (3). Using an inverted microscope with phase-contrast optics, micrographs were taken of the cultures in their live state and 1 h. after fixation with 2.5% glutaraldehyde in Dulbecco's phosphate buffered saline (Figs. 1 and 3).

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