Abstract
Many studies have been trying to establish standard protocols for animal experimentation, especially for animal species or strains, to master research variables with high precision. The main mouse strains used in the field of the biology of reproduction are Swiss, Balb/c, and C57BL/6. Since some of the strains show reproduction limitations, such as the size of the litter, the present study aimed to compare their spermatogenic processes to verify differences regarding the testicular parenchyma and germ cell populations, which could explain low offspring production. In addition, the present study provides additional information concerning the testicular parenchyma of such strains, which consequently would help researchers to choose the most suitable strain for reproductive studies. Six adult male mice were used for each of the strains. After euthanasia, the testes were weighed, fixated with Karnovsky fixative, embedded in methacrylate, sectioned, and stained with toluidine blue/sodium borate 1%. Morphometrical analyses from the testicular parenchyma (seminiferous tubules and interstitium) were made using the software ImageJ. Germ and Sertoli cells populations were counted in seminiferous tubules cross-sections at stage I of the seminiferous epithelium cycle. The lowest body and testicular weights were observed in C57BL/6 animals, followed by Balb/c and Swiss, however, the relative testes, parenchyma, and albuginea weights were significantly lower only in C57BL/6. Despite the seminiferous tubules and seminiferous epithelium proportions were lower in Swiss animals, their relative amount related to the body weight was the same among strains. The total number of germ cells was higher in Swiss animals, reflecting higher spermatogenic yield and daily sperm production. Due to the lower relative number of Sertoli cells, the Swiss animals showed the highest Sertoli cell index and support capacity. On the other hand, the lowest pathological indexes regarding the germ cells were observed in Balb/c animals, followed by Swiss and C57BL/6. In the interstitium, the proportion of blood vessels was lower in Swiss mice, while the lymphatic cell proportion was lower in C57BL/6 animals. Moreover, the highest proportions of Leydig cells and macrophages were noticed in Swiss mice, which may indicate increased testosterone levels. Altogether, such observations must be taken into account when choosing any of the studied strains for reproduction studies.
Highlights
The use of animal models for experimentation was responsible to improve methods of prevention, diagnosis, and treatment (Green 1986; Fernandes et al 2001; Almeida et al 2008)
The lowest body weight was observed in C57BL/6 animals, followed by Balb/c and Swiss
The testes weight, the GSI, the parenchyma weight and the albuginea weight were significantly lower in the C57BL/6 animals
Summary
The use of animal models for experimentation was responsible to improve methods of prevention, diagnosis, and treatment (Green 1986; Fernandes et al 2001; Almeida et al 2008). The use of nonhuman animals in research has significantly increased, allowing scientific advances such as the description of vital mechanisms, especially in medicine and physiology (Simmons 2004). Many studies have been trying to establish standard protocols, from chemical reagents to animal species or strains, to master the research variables with high precision, avoiding misinterpretation of the results, and increasing reliability (Spinelli et al 2012; Oliveira 2014). There is a need to establish the concept of the animal model, which would be the animal that best responds to the experimental approach as well as its reliable reproduction so that any researcher could have access to the same results (Maia et al 2006). The animal species must be capable of providing the appropriate answer to that specific experiment (Braga 2017)
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