Avaliação da técnica de coloração AgNOR em testículos de ovinos

Chacur, Marcelo George Mungai; Ibrahim, Daniel Boabaid; Arrebola, Thaís Andressa Hernandes; Sanches, Osimar Carvalho; Giuffrida, Rogério; Oba, Eunice; Ramos, Alcides de Amorim

doi:10.1590/1678-6695

Cited by 8 publications

(5 citation statements)

References 19 publications

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“…For this purpose, ovarian tissue fragments from the control and autotransplanted treatments were histologically processed, as described earlier. Thereafter, NOR labeling was performed and interpreted using an adapted protocol previously reported [ 49 ]. Briefly, after reduction with potassium iodide 1%, slides containing ovarian tissue sections of 5 μm (5 sections per ovary/per treatment) were stained with 50% silver nitrate solution in a colloid solution (2:1) in a darkroom and counterstained with 0.1% safranin.…”

Section: Methodsmentioning

confidence: 99%

Heterotopic autotransplantation of ovarian tissue in a large animal model: Effects of cooling and VEGF

Souza

Alves

et al. 2020

PLoS ONE

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Heterotopic and orthotopic ovarian tissue autotransplantation techniques, currently used in humans, will become promising alternative methods for fertility preservation in domestic and wild animals. Thus, this study describes for the first time the efficiency of a heterotopic ovarian tissue autotransplantation technique in a large livestock species (i.e., horses) after ovarian fragments were exposed or not to a cooling process (4°C/24 h) and/or VEGF before grafting. Ovarian fragments were collected in vivo via an ultrasound-guided biopsy pick-up method and surgically autografted in a subcutaneous site in both sides of the neck in each mare. The blood flow perfusion at the transplantation site was monitored at days 2, 4, 6, and 7 post-grafting using color-Doppler ultrasonography. Ovarian grafts were recovered 7 days post-transplantation and subjected to histological analyses. The exposure of the ovarian fragments to VEGF before grafting was not beneficial to the quality of the tissue; however, the cooling process of the fragments reduced the acute hyperemia post-grafting. Cooled grafts compared with non-cooled grafts contained similar values for normal and developing preantral follicles, vessel density, and stromal cell apoptosis; lower collagen type III fibers and follicular density; and higher stromal cell density, AgNOR, and collagen type I fibers. In conclusion, VEGF exposure before autotransplantation did not improve the quality of grafted tissues. However, cooling ovarian tissue for at least 24 h before grafting can be beneficial because satisfactory rates of follicle survival and development, stromal cell survival and proliferation, as well as vessel density, were obtained.

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Section: Methodsmentioning

confidence: 99%

Heterotopic autotransplantation of ovarian tissue in a large animal model: Effects of cooling and VEGF

Souza

Alves

et al. 2020

PLoS ONE

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“…The differences evidenced in the present study in the quantification of NORs in spermatogonia demonstrate that the choice of associations of cryoprotectants with lower cell toxicity, as DMSO/GLY, acts directly on the proliferative potential of the spermatogenic cells, as the higher the amount of NORs, the higher the possibility of spermatogonia to differentiate into sperm suitable to be used in assisted reproduction programs (Chacur et al., ).…”

Section: Discussionmentioning

confidence: 70%

“…For each fragment of each group evaluated, 10 cells were counted in 10 random fields, with a total count of 500 cells per experimental group. The counting of NOR was performed by observation of the black dots within the nucleoli of spermatogonia, at a 1000× magnification in an optical microscope (Chacur et al., ).…”

Section: Methodsmentioning

confidence: 99%

“…Furthermore, Chacur et al. () validated the AgNOR technique as an efficient method to identify NORs in spermatogonia and in Leiydig and Sertoli cells in fresh testicular fragments of sheep. However, there are no researches of any kind involving the use of this technique to assess the potential for proliferation of testicular tissue submitted to cryopreservation.…”

Section: Introductionmentioning

confidence: 99%

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Different associations of cryoprotectants for testicular tissue of prepubertal cats submitted to vitrification

Lima

Silva

Morais

et al. 2016

Reprod Domestic Animals

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The cryopreservation of testicular tissue is presented as the only alternative for the preservation of genetic material from prepubertal animals. However, this biotechnology is still being tested. The objective of this study was to evaluate the effect of different associations of cryoprotectants and the potential of cell proliferation after vitrification of testicular tissue of prepubertal cats. Five testicular pairs from five prepubertal cats were used, and each pair was divided into four fragments. Of these, one fragment composed of the control group (CG) and the rest were distributed in experimental groups according to the associations of cryoprotectants to be tested (dimethyl sulphoxide (DMSO)/glycerol (GLY); ethylene glycol (EG)/GLY) or DMSO/EG) in a final cryoprotectant concentration of 5.6 m. The fragments were submitted to vitrification, and after one week, fragments were heated and processed for histomorphological evaluation and quantification of nucleolar organizer regions (NORs). DMSO/GLY did not differ from CG and was superior to the other vitrified groups, as to cell separation and degree of shrinkage of the basal membrane. Concerning cell differentiation, visibility of the nucleus and nuclear condensation, all the vitrified groups were inferior to CG; however, DMSO/EG was inferior to DMSO/GLY and EG/GLY, which did not differ among themselves. CG was superior to all groups in quantification of NORs. DMSO/EG was inferior to all others, and there was no difference between DMSO/GLY and EG/GLY. The association DMSO/GLY presented the best preservation of tissue integrity and potential of cell proliferation after vitrification of the testicular tissue of prepubertal cats.

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“…Cell proliferation can be visualized by histochemical staining, such as the AgNORs method. Silver staining of nucleolus organizing regions (NORs) is characterized by marking ribosomal ribonucleic acid-bound proteins, assessing proliferation in normal or neoplastic cells (Chacur et al, 2015). Their count can be used to assess the rate of cell proliferation as these proteins tend to accumulate in mitotically active cells, while their expression in resting cells is low (Sirri et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

Morphological and Volumetric Characteristics of Holstein-Gir Crossbred Calves’ Stomachs Fed Diets Comprising Cheese Whey and Milk Powder

Miranda¹,

Teófilo²,

Assis³

et al. 2021

JSD

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The objective of this study was to evaluate the effect of powdered cheese whey and milk powder as a substitute for whole milk on rumen development in calves up to 60 days of age. Twenty-one-week-old Holstein-Gir crossbred calves were randomized across four treatments and six replicates: LI: whole milk (Control); LP: milk powder; LPS1: 80% milk powder + 20% cheese whey powder; LPS2: 60% Milk Powder + 40% Cheese Whey Powder. The animals were slaughtered at 60 days of age. The consumption, weight of stomach and its compartments, ruminal papilla height, and rumen proliferative activity were measured. Dry matter intake, absolute and relative weights of the gastric compartments were similar. Significant differences were found in the development of rumen papillae and the mean height of the ventral sac was higher in the animals that received whole milk. There were no significant differences for cell proliferation rate in rumen papillae. It is suggested that feeding calves whey with powdered milk may be a viable alternative to the use of whole milk without harming dairy calves.

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Avaliação da técnica de coloração AgNOR em testículos de ovinos

Cited by 8 publications

References 19 publications

Heterotopic autotransplantation of ovarian tissue in a large animal model: Effects of cooling and VEGF

Heterotopic autotransplantation of ovarian tissue in a large animal model: Effects of cooling and VEGF

Different associations of cryoprotectants for testicular tissue of prepubertal cats submitted to vitrification

Morphological and Volumetric Characteristics of Holstein-Gir Crossbred Calves’ Stomachs Fed Diets Comprising Cheese Whey and Milk Powder

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