Glycol Methacrylate Embedding for Improved Morphological, Morphometrical, and Immunohistochemical Investigations Under Light Microscopy: Testes as a Model

Chiarini-Garcia, Hélio; Parreira, Gleydes G.; Almeida, F. R. C. L.

doi:10.1007/978-1-60761-950-5_1

Cited by 22 publications

(14 citation statements)

References 25 publications

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“…After that, from the medial region of the ovaries, small fragments (~1–2 mm thickness) were obtained and, after ethanol dehydration, were embedded in glycol methacrylate resin (Historesin, Leica Microsystems Nussloch GmbH, Nussloch, Heidelberg, Germany). From the resin blocks, histological sections of 3 μm thickness were obtained and stained with toluidine blue‐sodium borate (Chiarini‐Garcia, Parreira, & Almeida, ) for histomorphometrical analysis.…”

Section: Methodsmentioning

confidence: 99%

Ovarian follicle development and genital tract characteristics in different birthweight gilts at 150 days of age

Almeida

Dias

Moreira

et al. 2017

Reprod Domestic Animals

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In the last decades, selection for improved prolificacy has resulted in higher litter sizes and has thereby increased the proportion of low birthweight (LW) piglets. It is well documented that LW piglets have lower growth performance, muscle accretion and poor carcass quality. However, little is known about the relations of birthweight with subsequent reproductive performance in gilts. This study investigated the effects of birthweight on reproductive tract and ovarian follicle development in 150-day-old gilts. Twenty eight female pigs of different birthweight ranges (high-HW: 1.8-2.2 kg; low-LW: 0.8-1.2 kg) from higher parity commercial sows were reared until 150 days of age, and their body weights were recorded at weaning, end of nursery and end of the grower-finisher phase. The animals were killed and their reproductive tracts collected for biometrical and histomorphometrical analysis. LW gilts showed significantly lower body weights and growth rates during all phases of production compared to their HW counterparts (p < .01). Most biometrical measurements of the reproductive tract were similar between the experimental groups, except vaginal length and the gonadossomatic index (relative ovarian weight), which were affected by birthweight class (p < .05). LW females also showed fewer medium size (3-5 mm; p < .01) ovarian follicles, pre-antral follicles (p < .07) and more atretic follicles per ovarian cortex area (p < .05). Therefore, besides the effects on post-natal growth performance, birthweight affects vaginal length and the follicular dynamics process, which may impair the reproductive performance of replacement gilts.

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Section: Methodsmentioning

confidence: 99%

Ovarian follicle development and genital tract characteristics in different birthweight gilts at 150 days of age

Almeida

Dias

Moreira

et al. 2017

Reprod Domestic Animals

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“…Before embedding in GMA, tissue fragments were cut to appropriate size for use in the KIT Historesin Leyca KIT (Leika Historesin, Embedding Kit, Heidelberg, Germany) as previously described. 20 Subsequently, tissues were dehydrated in absolute ethanol in three steps of 30 min each and two baths of xylene at 15 min each. After dehydration, the tissue samples were GMA-embedded using the Historesin Solution Kit.…”

Section: Methodsmentioning

confidence: 99%

“…The reported advantages compared to paraffin include: i) rapid processing; ii) ease of handling; iii) no requirement for high temperatures for infiltration and soluble polymerization assays; iv) ability to obtain semi-thin sections of 0.5 µm; v) less distortion and artifact; and vi) better resolution with light microscopy. 20 …”

Section: Introductionmentioning

confidence: 99%

Glycol methacrylate embedding for the histochemical study of the gastrointestinal tract of dogs naturally infected with Leishmania infantum

et al. 2015

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In canine visceral leishmaniasis a diffuse chronic inflammatory exudate and an intense parasite load throughout the gastrointestinal tract (GIT) has been previously reported. However, these studies did not allow a properly description of canine cellular morphology details. The aim of our study was to better characterize these cells in carrying out a qualitative and quantitative histological study in the gastrointestinal tract of dogs naturally infected with Leishmania infantum by examining gut tissues embedded in glycol methacrylate. Twelve infected adult dogs were classified in asymptomatic and symptomatic. Five uninfected dogs were used as controls. After necropsy, three samples of each gut segment, including oesophagus, stomach, duodenum, jejunum, ileum, cecum, colon, and rectum were collected and fixed in Carnoy’s solution for glycol methacrylate protocols. Sections were stained with hematoxylin-eosin, toluidine blue borate, and periodic acid-Schiff stain. Leishmania amastigotes were detected by immunohistochemistry employed in both glycol methacrylate and paraffin embedded tissues. The quantitative histological analysis showed higher numbers of plasma cells, lymphocytes and macrophages in lamina propria of all segments of GIT of infected dogs compared with controls. The parasite load was more intense and cecum and colon, independently of the clinical status of these dogs. Importantly, glycol methacrylate embedded tissue stained with toluidine blue borate clearly revealed mast cell morphology, even after mast cell degranulation. Infected dogs showed lower numbers of mast cells in all gut segments than controls. Despite the glycol methacrylate (GMA) protocol requires more attention and care than the conventional paraffin processing, this embedding procedure proved to be especially suitable for the present histological study, where it allowed to preserve and observe cell morphology in fine detail.

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“…Sections, 3–5 µm thick, were stained with toluidine blue‐sodium borate for general morphological and morphometric analyzes. Basement membranes arrangements were analyzed in 3–5µm thick sections stained with modified periodic acid Schiff – PAS (Chiarini‐Garcia et al, ).…”

Section: Methodsmentioning

confidence: 99%

Morphofunctional changes of female germinal epithelium to support spermatozoa along the annual reproductive cycle in an inseminating catfish (Trachelyopterus galeatus, Auchenipteridae)

Chiarini‐Garcia

Vieira

Godinho

2013

Journal of Morphology

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The reproductive system of some fish species presents elaborate mechanisms by which the females store spermatozoa inside their ovaries, keeping them viable for fertilization for an extended period of time. However, as intriguing as this sperm storage is, it is not yet understood how the sperm can remain viable in the ovary. Aiming to understand this phenomenon, the epithelium covering the ovarian lamellae, that is, the germinal epithelium, of the Cangati (Trachelyopterus galeatus), an inseminating catfish, was evaluated taking into account the different stages of the annual reproductive cycle. The germinal epithelium morphology changed during the annual reproductive cycle, presumably in preparation to receive the spermatozoa and keep them viable until fertilization. There was a progressive increase of the epithelium height. Also the number of intercellular junctions, desmosomes, and extended tight junctions, apparently increased forming chains that could be regarded as a barrier to isolate the sperm from the female immune system. Synthetic organelles were active releasing cytoplasmic granules and secretion in the epithelial enfolds in which the spermatozoa were deeply embedded. Concomitantly, oogonium nests were formed in the germinal epithelium during early folliculogenesis.

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Glycol Methacrylate Embedding for Improved Morphological, Morphometrical, and Immunohistochemical Investigations Under Light Microscopy: Testes as a Model

Cited by 22 publications

References 25 publications

Ovarian follicle development and genital tract characteristics in different birthweight gilts at 150 days of age

Ovarian follicle development and genital tract characteristics in different birthweight gilts at 150 days of age

Glycol methacrylate embedding for the histochemical study of the gastrointestinal tract of dogs naturally infected with Leishmania infantum

Morphofunctional changes of female germinal epithelium to support spermatozoa along the annual reproductive cycle in an inseminating catfish (Trachelyopterus galeatus, Auchenipteridae)

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