Transcriptome analysis of the spider Phonotimpus pennimani reveals novel toxin transcripts

Background: Phonotimpus pennimani (Araneae, Phrurolithidae) is a small-sized (3-5 mm) spider endemic to the Tacaná volcano in Chiapas, Mexico, where it is found in soil litter of cloud forests and coffee plantations. Its venom composition has so far not been investigated, partly because it is not a species of medical significance. However, it does have an important impact on the arthropod populations of its natural habitat. Methods: Specimens were collected in Southeastern Mexico (Chiapas) and identified taxonomically by morphological characteristics. A partial sequence from the mitochondrial gene coxI was amplified. Sequencing on the Illumina platform of a transcriptome library constructed from 12 adult specimens revealed 25 toxin or toxinlike genes. Transcripts were validated (RT-qPCR) by assessing the differential expression of the toxin-like PpenTox1 transcript and normalising with housekeeping genes. Results: Analysis of the coxI-gene revealed a similarity to other species of the family Phrurolithidae. Transcriptome analysis also revealed similarity with venom components of species from the families Ctenidae, Lycosidae, and Sicariidae. Expression of the toxinlike PpenTox1 gene was different for each developmental stage (juvenile or adult) and also for both sexes (female or male). Additionally, a partial sequence was obtained for the toxin-like PpenTox1 from DNA. Conclusion: Data from the amplification of the mitochondrial coxI gene confirmed that P. pennimani belongs to the family Phrurolithidae. New genes and transcripts coding for venom components were identified.(AU)

Analysis of nuclear maturation, DNA damage and repair gene expression of bovine oocyte and cumulus cells submitted to ionizing radiation'

Radiotherapy causes destruction of tumor cells, but also threatens the integrity and survival of surrounding normal cells. Then, woman submitted to irradiation for cancer treatment may present permanent ovary damage, resulting in impaired fertility. The objective of this study was to investigate the effects of therapeutic doses of ionizing radiation (IR), used for ovarian cancer treatment in humans, on bovine cumulus-oocyte complexes (COCs) as experimental model. Bovine ovaries were exposed to 0.9 Gy, 1.8 Gy, 3.6 Gy or 18.6 Gy IR, and then COCs were collected and used to evaluate: (a) oocyte nuclear maturation; (b) presence of phosphorylated H2A.X (γH2AX), as an indicator of DNA double-strand breaks (DSBs); and (c) expression of genes involved in DNA repair (TP53BP1, RAD52, ATM, XRCC6 and XRCC5) and apoptosis (BAX). The radiation doses tested in this study had no detrimental effects on nuclear maturation and did not increase γH2AX in the oocytes. However, IR treatment altered the mRNA abundance of RAD52 (RAD52 homolog, DNA repair protein) and BAX (BCL2-associated X protein). We conclude that although IR doses had no apparent effect on oocyte nuclear maturation and DNA damage, molecular pathways involved in DNA repair and apoptosis were affected by IR exposure in cumulus cells.(AU)

Expression analysis of transcription factors in sugarcane during cold stress / Análise de expressão de fatores de transcrição em cana-de-açúcar sob estresse por frio

Abstract Transcription factors (TF) are a wide class of genes in plants, and these can regulate the expression of other genes in response to various environmental stresses (biotic and abiotic). In the current study, transcription factor activity in sugarcane was examined during cold stress. Initially, RNA transcript reads of two sugarcane cultivars (ROC22 and GT08-1108) under cold stress were downloaded from SRA NCBI database. The reads were aligned into a reference genome and the differential expression analyses were performed with the R/Bioconductor edgeR package. Based on our analyses in the ROC22 cultivar, 963 TF genes were significantly upregulated under cold stress among a total of 5649 upregulated genes, while 293 TF genes were downregulated among a total of 3,289 downregulated genes. In the GT08-1108 cultivar, 974 TF genes were identified among 5,649 upregulated genes and 283 TF genes were found among 3,289 downregulated genes. Most transcription factors were annotated with GO categories related to protein binding, transcription factor binding, DNA-sequence-specific binding, transcription factor complex, transcription factor activity in RNA polymerase II, the activity of nucleic acid binding transcription factor, transcription corepressor activity, sequence-specific regulatory region, the activity of transcription factor of RNA polymerase II, transcription factor cofactor activity, transcription factor activity from plastid promoter, transcription factor activity from RNA polymerase I promoter, polymerase II and RNA polymerase III. The findings of above results will help to identify differentially expressed transcription factors during cold stress. It also provides a comprehensive analysis of the regulation of the transcription activity of many genes. Therefore, this study provides the molecular basis for improving cold tolerance in sugarcane and other economically important grasses.

Resumo Fatores de transcrição (FT) são uma ampla classe de genes em plantas e podem regular a expressão de outros genes em resposta a vários estresses ambientais (estresses bióticos e abióticos). No presente estudo, a atividade do fator de transcrição na cana-de-açúcar foi examinada durante o estresse pelo frio. Inicialmente, as leituras de transcrição de RNA de duas cultivares de cana-de-açúcar (ROC22 e GT08-1108) sob estresse frio foram baixadas do banco de dados SRA NCBI. As leituras foram alinhadas em um genoma de referência e as análises de expressão diferencial foram realizadas com o pacote R / Bioconductor edgeR. Com base em nossas análises no cultivar ROC22, 963 genes TF foram significativamente regulados positivamente sob estresse pelo frio entre um total de 5.649 genes regulados positivamente, enquanto 293 genes TF foram regulados negativamente entre um total de 3.289 genes regulados negativamente. No cultivar GT08-1108, 974 genes TF foram identificados entre 5.649 genes regulados positivamente e 283 genes TF foram encontrados entre 3.289 genes regulados negativamente. Os fatores de transcrição, em sua maioria, foram anotados com categorias GO relacionadas à ligação de proteína, ligação de fator de transcrição, ligação específica de sequência de DNA, complexo de fator de transcrição, atividade de fator de transcrição em RNA polimerase II, atividade de fator de transcrição de ligação de ácido nucleico, atividade de corepressor de transcrição, sequência específica da região reguladora, atividade do fator de transcrição da RNA polimerase II, atividade do cofator do fator de transcrição, atividade do fator de transcrição do promotor do plastídio, atividade do fator de transcrição do promotor da RNA polimerase I, polimerase II e RNA polimerase III. As descobertas dos resultados acima ajudarão a identificar fatores de transcrição expressos diferencialmente durante o estresse pelo frio. Ele também fornece uma análise abrangente da regulação da atividade de transcrição de muitos genes. Portanto, este estudo fornece base molecular para melhorar a tolerância ao frio em cana-de-açúcar e outras gramíneas economicamente importantes.

Expression analysis of transcription factors in sugarcane during cold stress

Abstract Transcription factors (TF) are a wide class of genes in plants, and these can regulate the expression of other genes in response to various environmental stresses (biotic and abiotic). In the current study, transcription factor activity in sugarcane was examined during cold stress. Initially, RNA transcript reads of two sugarcane cultivars (ROC22 and GT08-1108) under cold stress were downloaded from SRA NCBI database. The reads were aligned into a reference genome and the differential expression analyses were performed with the R/Bioconductor edgeR package. Based on our analyses in the ROC22 cultivar, 963 TF genes were significantly upregulated under cold stress among a total of 5649 upregulated genes, while 293 TF genes were downregulated among a total of 3,289 downregulated genes. In the GT08-1108 cultivar, 974 TF genes were identified among 5,649 upregulated genes and 283 TF genes were found among 3,289 downregulated genes. Most transcription factors were annotated with GO categories related to protein binding, transcription factor binding, DNA-sequence-specific binding, transcription factor complex, transcription factor activity in RNA polymerase II, the activity of nucleic acid binding transcription factor, transcription corepressor activity, sequence-specific regulatory region, the activity of transcription factor of RNA polymerase II, transcription factor cofactor activity, transcription factor activity from plastid promoter, transcription factor activity from RNA polymerase I promoter, polymerase II and RNA polymerase III. The findings of above results will help to identify differentially expressed transcription factors during cold stress. It also provides a comprehensive analysis of the regulation of the transcription activity of many genes. Therefore, this study provides the molecular basis for improving cold tolerance in sugarcane and other economically important grasses.

Resumo Fatores de transcrição (FT) são uma ampla classe de genes em plantas e podem regular a expressão de outros genes em resposta a vários estresses ambientais (estresses bióticos e abióticos). No presente estudo, a atividade do fator de transcrição na cana-de-açúcar foi examinada durante o estresse pelo frio. Inicialmente, as leituras de transcrição de RNA de duas cultivares de cana-de-açúcar (ROC22 e GT08-1108) sob estresse frio foram baixadas do banco de dados SRA NCBI. As leituras foram alinhadas em um genoma de referência e as análises de expressão diferencial foram realizadas com o pacote R / Bioconductor edgeR. Com base em nossas análises no cultivar ROC22, 963 genes TF foram significativamente regulados positivamente sob estresse pelo frio entre um total de 5.649 genes regulados positivamente, enquanto 293 genes TF foram regulados negativamente entre um total de 3.289 genes regulados negativamente. No cultivar GT08-1108, 974 genes TF foram identificados entre 5.649 genes regulados positivamente e 283 genes TF foram encontrados entre 3.289 genes regulados negativamente. Os fatores de transcrição, em sua maioria, foram anotados com categorias GO relacionadas à ligação de proteína, ligação de fator de transcrição, ligação específica de sequência de DNA, complexo de fator de transcrição, atividade de fator de transcrição em RNA polimerase II, atividade de fator de transcrição de ligação de ácido nucleico, atividade de corepressor de transcrição, sequência específica da região reguladora, atividade do fator de transcrição da RNA polimerase II, atividade do cofator do fator de transcrição, atividade do fator de transcrição do promotor do plastídio, atividade do fator de transcrição do promotor da RNA polimerase I, polimerase II e RNA polimerase III. As descobertas dos resultados acima ajudarão a identificar fatores de transcrição expressos diferencialmente durante o estresse pelo frio. Ele também fornece uma análise abrangente da regulação da atividade de transcrição de muitos genes. Portanto, este estudo fornece base molecular para melhorar a tolerância ao frio em cana-de-açúcar e outras gramíneas economicamente importantes.

Expression analysis of transcription factors in sugarcane during cold stress / Análise de expressão de fatores de transcrição em cana-de-açúcar sob estresse por frio

Transcription factors (TF) are a wide class of genes in plants, and these can regulate the expression of other genes in response to various environmental stresses (biotic and abiotic). In the current study, transcription factor activity in sugarcane was examined during cold stress. Initially, RNA transcript reads of two sugarcane cultivars (ROC22 and GT08-1108) under cold stress were downloaded from SRA NCBI database. The reads were aligned into a reference genome and the differential expression analyses were performed with the R/Bioconductor edgeR package. Based on our analyses in the ROC22 cultivar, 963 TF genes were significantly upregulated under cold stress among a total of 5649 upregulated genes, while 293 TF genes were downregulated among a total of 3,289 downregulated genes. In the GT08-1108 cultivar, 974 TF genes were identified among 5,649 upregulated genes and 283 TF genes were found among 3,289 downregulated genes. Most transcription factors were annotated with GO categories related to protein binding, transcription factor binding, DNA-sequence-specific binding, transcription factor complex, transcription factor activity in RNA polymerase II, the activity of nucleic acid binding transcription factor, transcription corepressor activity, sequence-specific regulatory region, the activity of transcription factor of RNA polymerase II, transcription factor cofactor activity, transcription factor activity from plastid promoter, transcription factor activity from RNA polymerase I promoter, polymerase II and RNA polymerase III. The findings of above results will help to identify differentially expressed transcription factors during cold stress. It also provides a comprehensive analysis of the regulation of the transcription activity of many genes. Therefore, this study provides the molecular basis for improving cold tolerance in sugarcane and other economically important grasses.

Fatores de transcrição (FT) são uma ampla classe de genes em plantas e podem regular a expressão de outros genes em resposta a vários estresses ambientais (estresses bióticos e abióticos). No presente estudo, a atividade do fator de transcrição na cana-de-açúcar foi examinada durante o estresse pelo frio. Inicialmente, as leituras de transcrição de RNA de duas cultivares de cana-de-açúcar (ROC22 e GT08-1108) sob estresse frio foram baixadas do banco de dados SRA NCBI. As leituras foram alinhadas em um genoma de referência e as análises de expressão diferencial foram realizadas com o pacote R / Bioconductor edgeR. Com base em nossas análises no cultivar ROC22, 963 genes TF foram significativamente regulados positivamente sob estresse pelo frio entre um total de 5.649 genes regulados positivamente, enquanto 293 genes TF foram regulados negativamente entre um total de 3.289 genes regulados negativamente. No cultivar GT08-1108, 974 genes TF foram identificados entre 5.649 genes regulados positivamente e 283 genes TF foram encontrados entre 3.289 genes regulados negativamente. Os fatores de transcrição, em sua maioria, foram anotados com categorias GO relacionadas à ligação de proteína, ligação de fator de transcrição, ligação específica de sequência de DNA, complexo de fator de transcrição, atividade de fator de transcrição em RNA polimerase II, atividade de fator de transcrição de ligação de ácido nucleico, atividade de corepressor de transcrição, sequência específica da região reguladora, atividade do fator de transcrição da RNA polimerase II, atividade do cofator do fator de transcrição, atividade do fator de transcrição do promotor do plastídio, atividade do fator de transcrição do promotor da RNA polimerase I, polimerase II e RNA polimerase III. As descobertas dos resultados acima ajudarão a identificar fatores de transcrição expressos diferencialmente durante o estresse pelo frio. Ele também fornece uma análise abrangente da regulação da atividade [...].

Expression analysis of transcription factors in sugarcane during cold stress / Análise de expressão de fatores de transcrição em cana-de-açúcar sob estresse por frio

Transcription factors (TF) are a wide class of genes in plants, and these can regulate the expression of other genes in response to various environmental stresses (biotic and abiotic). In the current study, transcription factor activity in sugarcane was examined during cold stress. Initially, RNA transcript reads of two sugarcane cultivars (ROC22 and GT08-1108) under cold stress were downloaded from SRA NCBI database. The reads were aligned into a reference genome and the differential expression analyses were performed with the R/Bioconductor edgeR package. Based on our analyses in the ROC22 cultivar, 963 TF genes were significantly upregulated under cold stress among a total of 5649 upregulated genes, while 293 TF genes were downregulated among a total of 3,289 downregulated genes. In the GT08-1108 cultivar, 974 TF genes were identified among 5,649 upregulated genes and 283 TF genes were found among 3,289 downregulated genes. Most transcription factors were annotated with GO categories related to protein binding, transcription factor binding, DNA-sequence-specific binding, transcription factor complex, transcription factor activity in RNA polymerase II, the activity of nucleic acid binding transcription factor, transcription corepressor activity, sequence-specific regulatory region, the activity of transcription factor of RNA polymerase II, transcription factor cofactor activity, transcription factor activity from plastid promoter, transcription factor activity from RNA polymerase I promoter, polymerase II and RNA polymerase III. The findings of above results will help to identify differentially expressed transcription factors during cold stress. It also provides a comprehensive analysis of the regulation of the transcription activity of many genes. Therefore, this study provides the molecular basis for improving cold tolerance in sugarcane and other economically important grasses.(AU)

Fatores de transcrição (FT) são uma ampla classe de genes em plantas e podem regular a expressão de outros genes em resposta a vários estresses ambientais (estresses bióticos e abióticos). No presente estudo, a atividade do fator de transcrição na cana-de-açúcar foi examinada durante o estresse pelo frio. Inicialmente, as leituras de transcrição de RNA de duas cultivares de cana-de-açúcar (ROC22 e GT08-1108) sob estresse frio foram baixadas do banco de dados SRA NCBI. As leituras foram alinhadas em um genoma de referência e as análises de expressão diferencial foram realizadas com o pacote R / Bioconductor edgeR. Com base em nossas análises no cultivar ROC22, 963 genes TF foram significativamente regulados positivamente sob estresse pelo frio entre um total de 5.649 genes regulados positivamente, enquanto 293 genes TF foram regulados negativamente entre um total de 3.289 genes regulados negativamente. No cultivar GT08-1108, 974 genes TF foram identificados entre 5.649 genes regulados positivamente e 283 genes TF foram encontrados entre 3.289 genes regulados negativamente. Os fatores de transcrição, em sua maioria, foram anotados com categorias GO relacionadas à ligação de proteína, ligação de fator de transcrição, ligação específica de sequência de DNA, complexo de fator de transcrição, atividade de fator de transcrição em RNA polimerase II, atividade de fator de transcrição de ligação de ácido nucleico, atividade de corepressor de transcrição, sequência específica da região reguladora, atividade do fator de transcrição da RNA polimerase II, atividade do cofator do fator de transcrição, atividade do fator de transcrição do promotor do plastídio, atividade do fator de transcrição do promotor da RNA polimerase I, polimerase II e RNA polimerase III. As descobertas dos resultados acima ajudarão a identificar fatores de transcrição expressos diferencialmente durante o estresse pelo frio. Ele também fornece uma análise abrangente da regulação da atividade [...].(AU)

Antral follicle count classification selects fertile heifers without influencing the carcass traits / A classificação da contagem folicular antral seleciona novilhas mais férteis sem influenciar as características da carcaça

This study aimed to evaluate the ovarian structure, estrus intensity, ultrasound carcass measurements, and pregnancy rate of Nelore breed heifers and cows in accordance with antral follicle counts (AFCs). We evaluated 503 heifers and 565 Nelore cows, with a mean age of 15.5±2.2 and 69.8±36.1 months, respectively, submitted to a fixed-time artificial insemination (FTAI) protocol. On day zero, all bovine females were examined using ultrasound to determine the AFC. The mean AFC of the heifers and cows were 20±8.6 and 22.5±8.4, respectively. The rib-eye area (REA) and fat thickness (FT) of the heifers (n = 119 for REA and n = 219 for FT) were measured using ultrasound imaging. The average conception rates at the first FTAI and at the end of the breeding season were 35.8% and 57.5%, respectively, for heifers and 45.1% and 78.9%, respectively, for cows. We demonstrated that the probability of pregnancy at the first FTAI and at the end of the breeding season for both young heifers and cows increased as the AFC decreased (P>0.001 and P=0.0123, respectively). FT and REA showed no correlation with AFC in heifers. The intensity of estrus expression was negatively correlated with AFC (−0.46; P<0.0001). In conclusion, Nelore heifers and cows with low AFC had a high probability of pregnancy during the entire breeding season. Thus, AFC can be used as a tool to select heifers with increased fertility without affecting carcass traits (REA and FT).

Este estudo teve como objetivo avaliar as estruturas ovarianas, intensidade do cio, medidas de carcaça por ultrassonografia e taxa de gestação de novilhas e vacas da raça Nelore com diferentes contagens de folículos antrais (CFA). Avaliamos 503 novilhas e 565 vacas Nelore, com idade média de 15,5 ± 2,2 e 69,8 ± 36,1 meses, respectivamente, submetidas a um protocolo de inseminação artificial em tempo fixo (IATF). No dia zero, todas as fêmeas foram examinadas por ultrassom para determinar a CFA. A CFA média das novilhas foi de 20 ± 8,6 e a das vacas foi de 22,5 ± 8,4 folículos. A área de olho de lombo (AOL) e a espessura de gordura (EG) das novilhas (n = 119 para AOL e n = 219 para EG) foram medidas utilizando imagens ultrassonográficas. As taxas médias de concepção na primeira IATF e no final da estação de monta foram de 35,8% e 57,5% e 45,1% e 78,9% para as novilhas e vacas, respectivamente. Demonstramos que a probabilidade de gestação na primeira IATF e no final da estação de monta, tanto para novilhas jovens quanto para vacas, foi maior quando a CFA era menor (P>0,001 e P=0,0123, respectivamente). A espessura da gordura e a AOL não mostraram nenhuma correlação com a CFA. A intensidade da expressão do cio foi negativamente correlacionada com o CFA (-0,46; P < 0,0001). Em conclusão, novilhas jovens e vacas da raça Nelore com menor CFA apresentam maior probabilidade de prenhez durante toda a estação de monta. Nenhuma relação entre a CFA e as características de carcaça foram observadas em novilhas Nelore. A CFA pode ser usada como uma ferramenta de seleção de novilhas com maior fertilidade sem influenciar as características de carcaça (AOL e EG).

Extraskeletal osteosarcoma in the duodenum of a dog

Background: Osteosarcoma is the most observed primary bone tumor in dogs, and may affect the appendicular and axial skeletons. In addition, it may be present in extraskeletal form, accounting for only 1% of cases. As shown by few reports in the literature, the involvement of the intestinal region by is rare. The objective of this study was to report the case of a 13-year-old Yorkshire dog, submitted to an exploratory laparotomy for suspected partial intestinal obstruction, diagnosed with extraskeletal osteosarcoma. Case: A 13-year-old dog, Yorkshire Terrier, male, presented clinical signs of gastrointestinal abnormalities. An ultrasound examination was performed and was found a mass in small intestine region with wall and lumen invasion. Then, was realized exploratory laparotomy and detected intestinal obstruction due to a mass with approximately 5.0 x 6.0 x 4.4 cm localized in duodenum. Surgical removal was performed and the sample sent to the veterinary diagnostic laboratory for histopathological examination. The sample had an irregular surface and firm consistency. In addition, when cut, the mass enveloped the intestinal layers and sometimes obstructed the lumen. Then, the sample were processed routinely for histopathology. After that, in microscopy evaluation was detected cell proliferation, affecting all layers of intestine. In detail, cells were elongated with pleomorphism marked and atypical mitosis. In addition, there was production of cartilage and bone matrix. So, due the absence of others sites, the neoplasm was considered primary of intestine. After that, to evaluate the expression of KI-67 and COX-2 was performed, and the cell proliferation index was 54.0% and the COX-2 expression was moderate in less than 10% of neoplastic cells. After the surgery, the patient was hospitalized for a week and continue the treatment in home. Afterwards, the tutor received the diagnosis, but even though he was instructed on the severity of the case, he chose not to undergo chemotherapy. After three months, the patient presented abdominal fluid and nodules in your liver, suggesting metastasis, but without diagnosis confirmation. The patient died five months after the diagnosis of extraskeletal osteosarcoma. However, no necropsy was realized, impossibility the diagnosis confirm. Discussion: The frequency of extraskeletal osteosarcoma in dogs remains unknown, with the mammary glands being the most affected site. In the present study, osteosarcoma affects the duodenal region and no reports of this neoplasm in the duodenum of dogs have been found in the literature. The clinical sign of dyschezia was important for the tutor to refer the animal to the veterinarian and perform the ultrasound in an attempt to elucidate the case, as the tumor mass is not always palpable. Histopathological examination and immunohistochemistry were necessary for the differential diagnosis and to establish the prognosis, although after the surgery the tutor chose not to perform chemotherapy. Extraskeletal osteosarcoma are usually highly metastatic, mainly affecting the lymph nodes and liver. In this case, the patient presented a liver nodule three months after the tumor removal surgery, but unfortunately, there was no diagnostic confirmation. Such neoplastic type is rarer and more aggressive than appendicular and axial osteosarcoma, with an average survival of 1 to 3 months. In this case, as a necropsy was not obtained, we cannot attribute the survival time to the disease. The survival rates of osteosarcomas in dogs are few months, but in the present case, although the patient died five months after surgery, the failure to perform a necropsy compromises the attribution of survival time to extraskeletal osteosarcoma.

Propolis anti-inflammatory effects on MAGE-1 and retinoic acid-treated dendritic cells and on Th1 and T regulatory cells

Background: Propolis exhibits huge potential in the pharmaceutical industry. In the present study, its effects were investigated on dendritic cells (DCs) stimulated with a tumor antigen (MAGE-1) and retinoic acid (RA) and on T lymphocytes to observe a possible differential activation of T lymphocytes, driving preferentially to Th1 or Treg cells. Methods: Cell viability, lymphocyte proliferation, gene expression (T-bet and FoxP3), and cytokine production by DCs (TNF-α, IL-10, IL-6 and IL-1ß) and lymphocytes (IFN-γ and TGF-ß) were analyzed. Results: MAGE-1 and RA alone or in combination with propolis inhibited TNF-α production and induced a higher lymphoproliferation compared to control, while MAGE1 + propolis induced IL-6 production. Propolis in combination with RA induced FoxP3 expression. MAGE-1 induced IFN-γ production while propolis inhibited it, returning to basal levels. RA inhibited TGF-ß production, what was counteracted by propolis. Conclusion: Propolis affected immunological parameters inhibiting pro-inflammatory cytokines and favoring the regulatory profile, opening perspectives for the control of inflammatory conditions.(AU)

Differential gene expression pattern and plasma sex steroids during testicular development in Genyatremus luteus (Perciforme: Haemulidae) (Bloch, 1790) / Padrão diferencial de expressão gênica e esteróides sexuais plasmáticos durante o desenvolvimento testicular em Genyatremus luteus (Pisces: Haemulidae) (Bloch, 1790)

The aim of the current study is to evaluate gene expression patterns of LH (lhr) and estrogen (er) receptors and plasma steroid levels during testicular development in Genyatremus luteus. Males were histologically classified as immature (n=7), maturing (n=7) and mature (n=7), based on the cellular structure of their testes. Plasma 11-KT concentration recorded peak at the final maturation stage. The highest plasma 17α-OHP concentrations were observed at the immature stage; they decreased at the maturation and mature stages. On the other hand, 17ß-estradiol (E2) recorded higher concentrations at the maturation stage. Er expression has significantly increased along the maturational development of animals' testes. The mRNA observed for the LH receptor has decreased from immature to maturing stage; it presented expression peak at the mature stage. There was high association between receptor gene expression and plasma steroid levels, mainly E2. The current study was the first to feature different reproductive maturation stages in male G. luteus specimens, based on cellular, endocrine and molecular aspects. In addition, it has shown that the gene expression profile for er and lhr receptors, as well as plasma 11-KT and E2 concentrations, are directly linked to testicular maturation, although they are not necessarily associated with the gonadosomatic index.

O objetivo deste estudo foi avaliar os padrões de expressão gênica dos receptores de LH (lhr) e de estrogênio (er) e dos níveis de esteróides plasmáticos durante o desenvolvimento testicular de Genyatremus luteus. Os machos foram classificados histologicamente em imaturos, em maturação e maduros, de acordo com a estrutura celular dos testículos. A concentração plasmática de 11-KT apresentou um pico na fase de maturação final (P<0.05). As maiores concentrações plasmáticas de 17α-OHP foram encontradas no estádio imaturo (P<0.05), com consequente diminuição nos estádios em maturação e maturo. O 17ß-estradiol (E2) apresentou maiores níveis de concentração no estádio em maturação (P<0.05). A expressão de er aumentou significativamente ao longo do desenvolvimento maturacional dos testículos (P<0.05). O mRNA para o receptor de LH diminuiu do estádio imaturo para o estádio em maturação (P<0.05) com consequente pico de expressão no estádio maduro. Houve alta relação entre a expressão gênica dos receptores e os níveis de esteróides plasmáticos, especialmente com E2. Em conclusão, este estudo caracterizou pela primeira vez, sob os aspectos celular, endócrino e molecular, os diferentes estádios de maturação reprodutiva em machos de G. luteus, demonstrando que o perfil da expressão gênica para os receptores er e lhr, bem como as concentrações plasmáticas de 11-KT e E2 foram diretamente relacionados à maturação testicular, apesar de não se relacionarem necessariamente com o índice gonadossomático.

Morfologia espermática em touros: quais os principais defeitos e o porquê ocorrem / Sperm morphology in bulls: which are the main defects and why they occur

A morfologia espermática é uma característica seminal que está associada à fertilidade e é, portanto, um componente importante na rotina do controle de qualidade em Centros de Coleta e Processamento de Sêmen (CCPS). É utilizada para caracterizar e quantificar os vários tipos de anormalidades nas células espermáticas do touro, usando preparação em montagem úmida para microscopia de Contraste de Interferência Diferencial (DIC), Contraste de Fase (CF) e Lâmina Corada (LC). O perfil da morfologia espermática permite a tomada de decisão sobre o aproveitamento ou descarte de um ejaculado para criopreservação e predizer a fertilidade de uma amostra de sêmen. Alterações no espermograma, com evidências do histórico e exame físico do touro, levam a interpretação da função testicular anormal, e consequentemente, ao diagnóstico, tratamento e prognóstico, se for o caso. As causas mais comuns de uma espermatogênese anormal nos touros incluem: termorregulação testicular anormal, desequilíbrios hormonais, estresse de manejo, ambiental e nutricional, efeitos de toxinas ou expressão de genes deletérios, além de defeitos de origem genética. Conhecendo-se a estrutura do espermatozoide, torna-se mais fácil a identificação das anormalidades espermáticas bem como a descrição das mesmas e uma melhor compreensão de sua origem.(AU)

Sperm morphology is a seminal trait that is associated with fertility and is therefore an important component of routine quality control in Semen Collection and Processing Centers (SCPC). It is used to characterize and quantify the various types of abnormalities in bull sperm cells, using wet mount preparation for Differential Interference Contrast (DIC), Phase Contrast (PC) and Slides Smear (SS) microscopy. The sperm morphology profile allows decision making on the use or disposal of an ejaculate for cryopreservation and predicts the fertility of a semen sample. Changes in sperm morphology may indicate that fertility may be impacted and therefore it is important to routinely monitor ejaculates. An abnormal spermogram with evidence from a history and physical examination of the bull can lead to the interpretation of abnormal testicular function, and consequently, to the diagnosis, treatment and prognosis, if any. The most common causes of abnormal spermatogenesis in bulls include: abnormal testicular thermoregulation, hormonal imbalances, management, environmental and nutritional stress, effects of toxins or expression of deleterious genes, in addition to defects of genetic origin. Knowledge in the structure of the sperm makes easier the identification of sperm abnormalities, as well as their description and provides a better understanding of their origin.(AU)

Hemoperitoneum in a cow with lumbar muscle hemangiosarcoma

Background: Hemangiosarcoma is a malignant neoplasm of endothelial cells with an infiltrative growth pattern. Hemangiosarcomas are frequently reported in canines and rare in felines, sheep, goats, swine, horses and cattle. Few cases of hemangiosarcoma were reported in cattle. In the present report, we describe the clinicopathological findings of a bovine muscle hemangiosarcoma. Case: A 6-year-old, Girolando cow from the Dairy Cattle Sector of the Federal Rural University of Rio de Janeiro (UFRRJ), Seropédica, presented sternal decubitus. Clinical signs were markedly pale mucous membranes, moderate dehydration, respiratory distress, and increased heart rate. The hematological examination revealed intense regenerative anemia. Due to the worsening of the clinical condition, the cow was submitted to euthanasia. The necropsy and collection of various fragments of organs were performed, which were sent to the "Setor de Anatomia Patológica" (SAP-UFRRJ). Tissues were fixed in 10 % buffered formalin, routinely processed for histology and stained with Hematoxylin and Eosin (HE). The external mucous membranes were markedly pale. Multifocal areas of 1.5 x 1.0 cm, irregular and dark red were observed dissecting the quadratus lumborum muscle (hemangiosarcoma) fibers. These neoplasms were associated with an extensive cruoric clot adhered to the muscle fibers. The extensive, red, friable mass measured approximately 76 x 55 x 20 cm on the serous surfaces of the organs of the peritoneal cavity (hemoperitoneum). The spleen was moderately reduced. The bone marrow was markedly pale. Histologically, it was observed that there was an extensive proliferation of endothelial cells in the quadratus lumbar muscle mass dissecting the epimysium and perimysium. Endothelial cells had moderate pleomorphism, organized in vascular channels and forming multifocally solid areas with a significant amount of eosinophilic fibrillar material (fibrin). Sections of muscle neoplasm were subjected to immunohistochemistry with anti-von Willebrand factor primary antibody, which showed a multifocal moderate cytoplasmic immunolabeling of neoplastic endothelial cells. Discussion: There are few reports of striated muscle hemangiosarcoma in cattle. Muscular hemangiosarcomas were reported in a 4-month-old calf in the left cervical trapezius muscle and a 6-year-old Holstein cow with left pelvic limb mass lateral and distal to the knee. Some reports presented hemangiosarcoma in the iliopsoas muscle, left cervical trapezius muscle, pelvic limb muscles and right cervical muscle of the bovine. In the presented report, hemoperitoneum occurred as a result of hemorrhages from muscle hemangiosarcoma. Other studies have demonstrated cavity hemorrhages in joint, pelvic, pleural and cranial cavities associated with hemangiosarcoma. Hemangiosarcoma with regenerative anemia must be distinguished from other diseases that cause anemia. The main differential diagnoses of bovine with anemia are vena cava syndrome, coumarin derivatives poisoning, acute poisoning by Pteridium spp., tick fever, anaplasmosis, babesiosis and trypanosomiasis. Hemangiosarcoma should be differentiated from other lesions as hemangioma, vascular tumor of lymphatic endothelium and perivascular wall tumors. Cases with poorly differentiated morphology should be submitted for immunohistochemistry. In the present hemangiosarcoma case, we have used the von Willebrand factor for immunohistochemistry diagnosis. Expression of angiogenic growth factors such as CD31, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and angiopoietin-1 (Ang-1) have also been used in the diagnosis of vascular proliferation lesions. Hemangiosarcoma in cattle should be included mainly in the differential diagnosis of diseases that cause acute anemia in cattle.

Nano-selenium and nano-zinc oxide supplementation in syrup on laying area, population size and hsp gene expression of honey bees in hot climate

This study was designed to examine the protective effects of nano-selenium and nano-zinc oxide on queen and workers performance under heat stress condition and gene expression of heat shock protein 70 (hsp70) as an index of heat tolerance. Sixty colonies were randomly assigned to five treatments with 12 replicates from June until early September. Sugar syrup (50%) containing no supplement or nano-selenium at levels of 50 and 100 µg L-1 or nano-zinc at levels of 100 and 200 µg L-1 was fed to colonies. Nano-selenium supplementations had no effect, but nano-zinc at level of 100 µg L-1 significantly decreased body malondialdehyde concentration. The highest bee population was seen in nano-zinc at level of 100 µg L-1 and the lowest one in control group. The lowest and the highest body weight, fat and protein deposition was found in group received nano-zinc at level of 100 µg L-1 and control, respectively. The highest gene expression was for group received nano-zinc at level of 100 µg L-1 In group received nano-zinc at level of 100 µg L-1, an increase in hsp70 gene expression was found. In conclusion, nano-zinc oxide at level of 100 µg L-1 could increase queen and worker performance and heat resistance of bees in the hot climate condition.(AU)

Nano-selenium and nano-zinc oxide supplementation in syrup on laying area, population size and hsp gene expression of honey bees in hot climate

This study was designed to examine the protective effects of nano-selenium and nano-zinc oxide on queen and workers performance under heat stress condition and gene expression of heat shock protein 70 (hsp70) as an index of heat tolerance. Sixty colonies were randomly assigned to five treatments with 12 replicates from June until early September. Sugar syrup (50%) containing no supplement or nano-selenium at levels of 50 and 100 µg L-1 or nano-zinc at levels of 100 and 200 µg L-1 was fed to colonies. Nano-selenium supplementations had no effect, but nano-zinc at level of 100 µg L-1 significantly decreased body malondialdehyde concentration. The highest bee population was seen in nano-zinc at level of 100 µg L-1 and the lowest one in control group. The lowest and the highest body weight, fat and protein deposition was found in group received nano-zinc at level of 100 µg L-1 and control, respectively. The highest gene expression was for group received nano-zinc at level of 100 µg L-1 In group received nano-zinc at level of 100 µg L-1, an increase in hsp70 gene expression was found. In conclusion, nano-zinc oxide at level of 100 µg L-1 could increase queen and worker performance and heat resistance of bees in the hot climate condition.

Possibilidades do uso do RNA espermático como ferramenta na suinocultura / Possibilities of using sperm's RNA as a tool in pig farming

A seleção de macho mais férteis é essencial para a suinocultura moderna, a análise do transcriptoma espermático tem se mostrado como uma ferramenta interessante para podermos escolher animais com maior potencial de fertilidade ou congelabilidade de sêmen. O objetivo da presente revisão foi caracterizar os RNA encontrados em espermatozoides suínos e apresentar possíveis finalidades de seu uso na suinocultura. Por sofrer mudanças estruturais significativas, como meiose, compactação do DNA e perda de parte do citoplasma, as linhagens celulares que dão origem aos espermatozoides também apresentam uma população singular na população RNA. Os espermatozoides carregam um transcriptoma heterogêneo, com mRNA e muitos pequenos RNA truncados e de difícil avaliação. Eventos como congelamento, capacitação e estresse térmico causam alteração nos perfis de RNA espermático. Existem RNA que podemos relacionar como marcadores moleculares para diversas funções, além disso, há muitas evidências de que o RNA espermático contribua na modulação gênica durante a fertilização e desenvolvimento embrionário inicial, tornando estes em um importante alvo para estudos futuros.

The selection of fertile males is essential for modern pig farming, the sperm transcriptome analysis has been shown to be an interesting tool to be able to choose animals with greater fertility potential or semen freezeability. The aim of this review was to characterize the RNA found in boar spermatozoa and to present possible purposes for its use in pig farming. By undergoing significant structural changes, such as meiosis, DNA compaction and loss of part of the cytoplasm, spermatic cells also have a unique population in the RNA population. Sperm carry a heterogeneous transcriptome, with mRNA and many small and truncated RNAs. Events such as freezing, capacitation and heat stress cause changes in sperm RNA profiles. There are RNAs that we can relate as molecular markers for several functions, in addition, there is evidence that sperm RNA contributes to gene modulation during fertilization and early embryonic development, making these an important target for future studies.

Cell differentiation events in pre-implantation mouse and bovine embryos

Abstract Early mammal embryogenesis starts with oocyte fertilization, giving rise to the zygote. The events that the newly formed zygote surpasses are crucial to the embryo developmental success. Shortly after activation of its genome, cells of the embryo segregate into the inner cell mass (ICM) or the trophectoderm (TE). The first will give rise to the embryo while the latter will become the placenta. This first segregation involves cellular and molecular processes that include cell polarity linked to intracellular pathway activation, which will regulate the transcription of trophectoderm-related genes. Then, cells of the ICM undergo the second event of mammalian cell differentiation, which consists of the separation between epiblast (EPI) and hypoblast or primitive endoderm (PrE). This second segregation involves paracrine signaling, leading to differential expression of key genes that will dictate the fate of the cell. Although these processes are described in detail in the mouse, recent studies suggest that the bovine embryo could also be an interesting model for early development, since there are differences to the mouse and similarities with early human embryogenesis. In this review, we gathered the main data available in the literature upon bovine and mouse early development events, suggesting that both models should be analyzed and studied in a complementary way, to better model early events occurring in human development.

Evaluation of interferon-stimulated gene 15 (ISG15) expression in blood neutrophils in beef cattle poisoned by Senecio spp / Avaliação da expressão do gene estimulado por interferon 15 (ISG15) em neutrófilos sanguíneos em bovinos de corte intoxicados por Senecio spp

This study aimed to assess liver damage and interferon-stimulated gene 15 (ISG15) blood expression as a consequence of embryonic signaling on maternal recognition of pregnancy in beef cattle presenting natural ingestion of Senecio spp. Epidemiological aspects, as the presence of the plant, associated to gamma glutamyl transferase (GGT) activity can be used as Senecio spp. poisoning diagnosis. Maternal recognition of pregnancy period occurs when the embryo secretes interferon tau (IFNT) to signal its presence to the mother and eventually extend corpus luteum (CL) lifespan. In our study, liver damage was determined by concentration serum GGT, cytological and histopathological examinations. Reproductive status was evaluated by concentration of progesterone, CL diameter and ISG15 mRNA expression on Day 19 following fixed-time artificial insemination (FTAI). Cows were categorized into two groups based on concentration of GGT: Group 1 (GGT<30U/L) and 2 (GGT>31U/L). No difference on body condition scores was observed. All the cows presented liver damage based on cytology and histopathological exams. Cows from the Group 1 had higher pregnancy rate, presenting larger CL diameter and greater concentration of progesterone. Interestingly, ISG15 mRNA expression had no difference between Groups 1 and 2, even presenting difference in pregnancy status. These findings suggest embryonic loss beyond Day 19. It suggests late embryonic mortality may be associated to liver insufficiency. In conclusion, liver injury and/or concentration of GGT does not alter ISG15 expression on blood neutrophils, however cows presenting lower concentration of GGT (<30U/L) had increased pregnancy status. Therefore, the concentration of GGT allow us to screen liver status and foresee a successful pregnancy in beef cattle.(AU)

O objetivo deste estudo foi avaliar a lesão hepática e a expressão sanguínea do gene estimulado por interferon 15 (ISG15) durante a sinalização embrionária, no reconhecimento materno da gestação, em bovinos de corte apresentando ingestão natural de Senecio spp. Fatores epidemiológicos, como a presença da planta, associados à atividade da gama glutamil transferase (GGT) podem ser utilizados como diagnóstico da intoxicação por Senecio spp. O reconhecimento materno da gestação ocorre quando o embrião secreta interferon tau (IFNT) para sinalizar sua presença à mãe. Em nosso estudo, a lesão hepática foi determinada pela concentração sérica de GGT, pelos exames citológicos e histopatológicos. O estado reprodutivo foi avaliado pela concentração de progesterona, diâmetro de corpo lúteo (CL) e expressão de mRNA ISG15 no Dia 19 após a inseminação artificial em tempo fixo (IATF). As vacas foram separadas em dois grupos com base na concentração de GGT sanguíneo: Grupo 1 (GGT<30U/L) e Grupo 2 (GGT>31U/L). Não foi observada nenhuma diferença no escore de condição corporal entre os grupos. Na citologia e nos exames histopatológicos todas as vacas apresentaram lesão hepática. As vacas do Grupo 1 apresentaram maior taxa de prenhez, maior diâmetro do CL e maior concentração de progesterona. Diferente do esperado, a expressão do mRNA ISG15 não foi diferente entre os Grupos 1 e 2, mesmo apresentando diferença na taxa de prenhez. Esses achados sugerem perda embrionária após o Ddia 19. Isso demonstra que a mortalidade embrionária tardia pode estar associada à insuficiência hepática. Dessa forma, conclui-se que a lesão hepática e/ou concentração de GGT não altera a expressão de ISG15 nos neutrófilos sanguíneos, porém vacas com menor concentração de GGT (<30U/L) apresentaram maiores taxas de prenhez. Assim, a concentração de GGT nos permite avaliar a saúde hepática e prever uma gestação bem-sucedida em bovinos de corte.(AU)

Evaluation of interferon-stimulated gene 15 (ISG15) expression in blood neutrophils in beef cattle poisoned by Senecio spp / Avaliação da expressão do gene estimulado por interferon 15 (ISG15) em neutrófilos sanguíneos em bovinos de corte intoxicados por Senecio spp

This study aimed to assess liver damage and interferon-stimulated gene 15 (ISG15) blood expression as a consequence of embryonic signaling on maternal recognition of pregnancy in beef cattle presenting natural ingestion of Senecio spp. Epidemiological aspects, as the presence of the plant, associated to gamma glutamyl transferase (GGT) activity can be used as Senecio spp. poisoning diagnosis. Maternal recognition of pregnancy period occurs when the embryo secretes interferon tau (IFNT) to signal its presence to the mother and eventually extend corpus luteum (CL) lifespan. In our study, liver damage was determined by concentration serum GGT, cytological and histopathological examinations. Reproductive status was evaluated by concentration of progesterone, CL diameter and ISG15 mRNA expression on Day 19 following fixed-time artificial insemination (FTAI). Cows were categorized into two groups based on concentration of GGT: Group 1 (GGT<30U/L) and 2 (GGT>31U/L). No difference on body condition scores was observed. All the cows presented liver damage based on cytology and histopathological exams. Cows from the Group 1 had higher pregnancy rate, presenting larger CL diameter and greater concentration of progesterone. Interestingly, ISG15 mRNA expression had no difference between Groups 1 and 2, even presenting difference in pregnancy status. These findings suggest embryonic loss beyond Day 19. It suggests late embryonic mortality may be associated to liver insufficiency. In conclusion, liver injury and/or concentration of GGT does not alter ISG15 expression on blood neutrophils, however cows presenting lower concentration of GGT (<30U/L) had increased pregnancy status. Therefore, the concentration of GGT allow us to screen liver status and foresee a successful pregnancy in beef cattle.(AU)

O objetivo deste estudo foi avaliar a lesão hepática e a expressão sanguínea do gene estimulado por interferon 15 (ISG15) durante a sinalização embrionária, no reconhecimento materno da gestação, em bovinos de corte apresentando ingestão natural de Senecio spp. Fatores epidemiológicos, como a presença da planta, associados à atividade da gama glutamil transferase (GGT) podem ser utilizados como diagnóstico da intoxicação por Senecio spp. O reconhecimento materno da gestação ocorre quando o embrião secreta interferon tau (IFNT) para sinalizar sua presença à mãe. Em nosso estudo, a lesão hepática foi determinada pela concentração sérica de GGT, pelos exames citológicos e histopatológicos. O estado reprodutivo foi avaliado pela concentração de progesterona, diâmetro de corpo lúteo (CL) e expressão de mRNA ISG15 no Dia 19 após a inseminação artificial em tempo fixo (IATF). As vacas foram separadas em dois grupos com base na concentração de GGT sanguíneo: Grupo 1 (GGT<30U/L) e Grupo 2 (GGT>31U/L). Não foi observada nenhuma diferença no escore de condição corporal entre os grupos. Na citologia e nos exames histopatológicos todas as vacas apresentaram lesão hepática. As vacas do Grupo 1 apresentaram maior taxa de prenhez, maior diâmetro do CL e maior concentração de progesterona. Diferente do esperado, a expressão do mRNA ISG15 não foi diferente entre os Grupos 1 e 2, mesmo apresentando diferença na taxa de prenhez. Esses achados sugerem perda embrionária após o Ddia 19. Isso demonstra que a mortalidade embrionária tardia pode estar associada à insuficiência hepática. Dessa forma, conclui-se que a lesão hepática e/ou concentração de GGT não altera a expressão de ISG15 nos neutrófilos sanguíneos, porém vacas com menor concentração de GGT (<30U/L) apresentaram maiores taxas de prenhez. Assim, a concentração de GGT nos permite avaliar a saúde hepática e prever uma gestação bem-sucedida em bovinos de corte.(AU)

Histopathological evaluation and expression profile of PRM-1, TNP-2, 17B-HSD3, LHR, MHC-I, MIC-B, NC1 and NC3 genes in bovine testis / Análise histopatológica e perfil de expressão dos genes PRM-1, TNP-2, 17B-HSD3, LHR, MHC-I, MIC-B, NC1 e NC3 em testículo bovino

Histopathological and spermatogenesis classification by Johnsen is widely used in the germinal epithelium maturation analysis, besides identifying pathological alterations able to cause subfertility and even infertility. The aim of this study was to analyze cell-differentiation histopathological data and to correlate them with expression of PRM-1, TNP-2, 17β-HSD3, LHR, generic MHC-I, MIC-B, NC1 and NC3 genes, involved in bovine spermatogenesis using qRT-PCR from testicular parenchyma. Based on Johnsen’s criteria, the results showed normal spermatogenic activity in these animals, classified at 6, 7 and 8 scores. The qRT-PCR analysis expression showed that MHC-I (generic) gene was less expressed than all the other genes in evaluated scores (p < 0.05) and, PRM-1 and TNP-2 were the most expressed genes (p < 0.05). The PRM-1 gene expression was significantly higher than TNP-2 (p < 0.05). Comparing scores, 17β-HSD3 gene expression was lower (p < 0.05) in score 6 when compared to scores 7 and 8 animals. It was also observed that PRM-1 expression was lower in score 6 when compared to 7, as well as TNP-2 gene was less expressed in the score 6 (p < 0.05) when compared to 7 and 8 scores. Our results demonstrated that MHC I (generic), MIC-B, NC1, NC3, and LHR genes are poorly expressed in bovine testis, suggesting their marginal action on spermatogenesis. Instead, PRM-1, TNP-2, and 17β-HSD3 expression were higher, supporting the notion that these genes can act directly on the germ cells differential development during bovine spermatogenesis.

A classificação histopatológica e espermatogênica segundo Johnsen é amplamente utilizada na análise da matu-ração do epitélio germinativo, além de identificar alterações patológicas capazes de causar subfertilidade e até infertilidade. O objetivo deste estudo foi analisar dados histopatológicos de diferenciação celular e correlacioná-los à expressão dos genes PRM-1, TNP-2, 17β-HSD3, LHR, MHC-I genérico, MIC-B, NC1 e NC3, envolvidos na espermatogênese bovina usando qRT-PCR do parênquima testicular. Com base nos critérios de Johnsen, os resultados demonstraram atividade espermatogê-nica normal nesses animais, classificados nos escores 6, 7 e 8. A análise de expressão gênica (qRT-PCR) demonstrou que o gene MHC-I (genérico) foi menos expresso do que os demais nos escores avaliados (p < 0,05) e, PRM-1 e TNP-2 foram os mais expressos (p < 0,05). A expressão do PRM-1 foi significativamente maior do que TNP-2 (p < 0,05). Comparando os escores, a expressão do 17β-HSD3 foi menor (p < 0,05) no escore 6 quando comparado aos escores 7 e 8. Observou-se também que a expressão do PRM-1 foi menor no escore 6 quando comparado ao 7, bem como o TNP-2 foi menos expresso no escore 6 (p < 0,05) quando comparado aos escores 7 e 8. Nossos resultados demonstraram que os genes MHC-I (genérico), MIC-B, NC1, NC3 e LHR são pouco expressos em testículos bovinos, sugerindo sua ação indireta na espermatogênese. Em contrapartida, a expressão de PRM-1, TNP-2 e 17β-HSD3 foi alta, corroborando com a hipótese de que esses genes podem atuar diretamente no desenvolvimento diferencial de células germinativas durante a espermatogênese bovina.

Histopathological evaluation and expression profile of PRM-1, TNP-2, 17B-HSD3, LHR, MHC-I, MIC-B, NC1 and NC3 genes in bovine testis / Análise histopatológica e perfil de expressão dos genes PRM-1, TNP-2, 17B-HSD3, LHR, MHC-I, MIC-B, NC1 e NC3 em testículo bovino

Histopathological and spermatogenesis classification by Johnsen is widely used in the germinal epithelium maturation analysis, besides identifying pathological alterations able to cause subfertility and even infertility. The aim of this study was to analyze cell-differentiation histopathological data and to correlate them with expression of PRM-1, TNP-2, 17β-HSD3, LHR, generic MHC-I, MIC-B, NC1 and NC3 genes, involved in bovine spermatogenesis using qRT-PCR from testicular parenchyma. Based on Johnsens criteria, the results showed normal spermatogenic activity in these animals, classified at 6, 7 and 8 scores. The qRT-PCR analysis expression showed that MHC-I (generic) gene was less expressed than all the other genes in evaluated scores (p < 0.05) and, PRM-1 and TNP-2 were the most expressed genes (p < 0.05). The PRM-1 gene expression was significantly higher than TNP-2 (p < 0.05). Comparing scores, 17β-HSD3 gene expression was lower (p < 0.05) in score 6 when compared to scores 7 and 8 animals. It was also observed that PRM-1 expression was lower in score 6 when compared to 7, as well as TNP-2 gene was less expressed in the score 6 (p < 0.05) when compared to 7 and 8 scores. Our results demonstrated that MHC I (generic), MIC-B, NC1, NC3, and LHR genes are poorly expressed in bovine testis, suggesting their marginal action on spermatogenesis. Instead, PRM-1, TNP-2, and 17β-HSD3 expression were higher, supporting the notion that these genes can act directly on the germ cells differential development during bovine spermatogenesis.(AU)

A classificação histopatológica e espermatogênica segundo Johnsen é amplamente utilizada na análise da matu-ração do epitélio germinativo, além de identificar alterações patológicas capazes de causar subfertilidade e até infertilidade. O objetivo deste estudo foi analisar dados histopatológicos de diferenciação celular e correlacioná-los à expressão dos genes PRM-1, TNP-2, 17β-HSD3, LHR, MHC-I genérico, MIC-B, NC1 e NC3, envolvidos na espermatogênese bovina usando qRT-PCR do parênquima testicular. Com base nos critérios de Johnsen, os resultados demonstraram atividade espermatogê-nica normal nesses animais, classificados nos escores 6, 7 e 8. A análise de expressão gênica (qRT-PCR) demonstrou que o gene MHC-I (genérico) foi menos expresso do que os demais nos escores avaliados (p < 0,05) e, PRM-1 e TNP-2 foram os mais expressos (p < 0,05). A expressão do PRM-1 foi significativamente maior do que TNP-2 (p < 0,05). Comparando os escores, a expressão do 17β-HSD3 foi menor (p < 0,05) no escore 6 quando comparado aos escores 7 e 8. Observou-se também que a expressão do PRM-1 foi menor no escore 6 quando comparado ao 7, bem como o TNP-2 foi menos expresso no escore 6 (p < 0,05) quando comparado aos escores 7 e 8. Nossos resultados demonstraram que os genes MHC-I (genérico), MIC-B, NC1, NC3 e LHR são pouco expressos em testículos bovinos, sugerindo sua ação indireta na espermatogênese. Em contrapartida, a expressão de PRM-1, TNP-2 e 17β-HSD3 foi alta, corroborando com a hipótese de que esses genes podem atuar diretamente no desenvolvimento diferencial de células germinativas durante a espermatogênese bovina.(AU)